IL-2_NN gene_NN expression_NN and_CC NF-kappa_NN B_NN activation_NN through_IN CD28_NN requires_VBZ reactive_JJ oxygen_NN production_NN by_IN 5-lipoxygenase_NN ._. 
Activation_NN of_IN the_DT CD28_NN surface_NN receptor_NN provides_VBZ a_DT major_JJ costimulatory_JJ signal_NN for_IN T_NN cell_NN activation_NN resulting_VBG in_IN enhanced_VBN production_NN of_IN interleukin-2_NN (_( IL-2_NN )_) and_CC cell_NN proliferation_NN ._. 
In_IN primary_JJ T_NN lymphocytes_NNS we_PRP show_VBP that_IN CD28_NN ligation_NN leads_VBZ to_TO the_DT rapid_JJ intracellular_JJ formation_NN of_IN reactive_JJ oxygen_NN intermediates_NNS (_( ROIs_NNS )_) which_WDT are_VBP required_VBN for_IN CD28_* -mediated_JJ activation_NN of_IN the_DT NF-kappa_NN B_* /_* CD28_* -responsive_JJ complex_NN and_CC IL-2_NN expression_NN ._. 
Delineation_NN of_IN the_DT CD28_NN signaling_NN cascade_NN was_VBD found_VBN to_TO involve_VB protein_NN tyrosine_NN kinase_NN activity_NN ,_, followed_VBN by_IN the_DT activation_NN of_IN phospholipase_NN A2_NN and_CC 5-lipoxygenase_NN ._. 
Our_PRP$ data_NNS suggest_VBP that_IN lipoxygenase_NN metabolites_NNS activate_VBP ROI_NN formation_NN which_WDT then_RB induce_VBP IL-2_NN expression_NN via_IN NF-kappa_NN B_NN activation_NN ._. 
These_DT findings_NNS should_MD be_VB useful_JJ for_IN therapeutic_JJ strategies_NNS and_CC the_DT development_NN of_IN immunosuppressants_NNS targeting_VBG the_DT CD28_NN costimulatory_NN pathway_NN ._. 
The_DT peri-kappa_NN B_NN site_NN mediates_VBZ human_JJ immunodeficiency_NN virus_NN type_NN 2_CD enhancer_NN activation_NN in_IN monocytes_NNS but_CC not_RB in_IN T_NN cells_NNS ._. 
Human_JJ immunodeficiency_NN virus_NN type_NN 2_CD (_( HIV-2_NN )_) ,_, like_IN HIV-1_NN ,_, causes_VBZ AIDS_NN and_CC is_VBZ associated_VBN with_IN AIDS_NN cases_NNS primarily_RB in_IN West_NNP Africa_NNP ._. 
HIV-1_NN and_CC HIV-2_NN display_VBP significant_JJ differences_NNS in_IN nucleic_JJ acid_NN sequence_NN and_CC in_IN the_DT natural_JJ history_NN of_IN clinical_JJ disease_NN ._. 
Consistent_JJ with_IN these_DT differences_NNS ,_, we_PRP have_VBP previously_RB demonstrated_VBN that_IN the_DT enhancer/promoter_NN region_NN of_IN HIV-2_NN functions_VBZ quite_RB differently_RB from_IN that_DT of_IN HIV-1_NN ._. 
Whereas_IN activation_NN of_IN the_DT HIV-1_NN enhancer_NN following_VBG T-cell_NN stimulation_NN is_VBZ mediated_VBN largely_RB through_IN binding_NN of_IN the_DT transcription_NN factor_NN NF-kappa_NN B_NN to_TO two_CD adjacent_JJ kappa_NN B_NN sites_NNS in_IN the_DT HIV-1_NN long_JJ terminal_JJ repeat_NN ,_, activation_NN of_IN the_DT HIV-2_NN enhancer_NN in_IN monocytes_NNS and_CC T_NN cells_NNS is_VBZ dependent_JJ on_IN four_CD cis-acting_JJ elements_NNS :_: a_DT single_JJ kappa_NN B_NN site_NN ,_, two_CD purine-rich_JJ binding_VBG sites_NNS ,_, PuB1_NN and_CC PuB2_NN ,_, and_CC a_DT pets_NN site_NN ._. 
We_PRP have_VBP now_RB identified_VBN a_DT novel_JJ cis-acting_JJ element_NN within_IN the_DT HIV-2_NN enhancer_NN ,_, immediately_RB upstream_JJ of_IN the_DT kappa_NN B_NN site_NN ,_, designated_VBN peri-kappa_NN B_NN ._. 
This_DT site_NN is_VBZ conserved_VBN among_IN isolates_NNS of_IN HIV-2_NN and_CC the_DT closely_RB related_JJ simian_JJ immunodeficiency_NN virus_NN ,_, and_CC transfection_NN assays_NNS show_VBP this_DT site_NN to_TO mediate_VB HIV-2_NN enhancer_NN activation_NN following_VBG stimulation_NN of_IN monocytic_JJ but_CC not_RB T-cell_NN lines_NNS ._. 
This_DT is_VBZ the_DT first_JJ description_NN of_IN an_DT HIV-2_NN enhancer_NN element_NN which_WDT displays_VBZ such_JJ monocyte_NN specificity_NN ,_, and_CC no_DT comparable_JJ enhancer_NN element_NN has_VBZ been_VBN clearly_RB defined_VBN for_IN HIV-1_NN ._. 
While_IN a_DT nuclear_JJ factor_NN (_( s_NNS )_) from_IN both_CC peripheral_JJ blood_NN monocytes_NNS and_CC T_NN cells_NNS binds_VBZ the_DT peri-kappa_NN B_NN site_NN ,_, electrophoretic_JJ mobility_NN shift_NN assays_NNS suggest_VBP that_IN either_CC a_DT different_JJ protein_NN binds_VBZ to_TO this_DT site_NN in_IN monocytes_NNS versus_CC T_NN cells_NNS or_CC that_IN the_DT protein_NN recognizing_VBG this_DT enhancer_NN element_NN undergoes_VBZ differential_JJ modification_NN in_IN monocytes_NNS and_CC T_NN cells_NNS ,_, thus_RB supporting_VBG the_DT transfection_NN data_NNS ._. 
Further_RB ,_, while_IN specific_JJ constitutive_JJ binding_NN to_TO the_DT peri-kappa_NN B_NN site_NN is_VBZ seen_VBN in_IN monocytes_NNS ,_, stimulation_NN with_IN phorbol_NN esters_NNS induces_VBZ additional_JJ ,_, specific_JJ binding_NN ._. 
Understanding_VBG the_DT monocyte-specific_JJ function_NN of_IN the_DT peri-kappa_NN B_NN factor_NN may_MD ultimately_RB provide_VB insight_NN into_IN the_DT different_JJ role_NN monocytes_NNS and_CC T_NN cells_NNS play_VBP in_IN HIV_NN pathogenesis_NN ._. 
E1A_NN gene_NN expression_NN induces_VBZ susceptibility_NN to_TO killing_NN by_IN NK_NN cells_NNS following_VBG immortalization_NN but_CC not_RB adenovirus_NN infection_NN of_IN human_JJ cells_NNS ._. 
Adenovirus_NN (_( Ad_NN )_) infection_NN and_CC E1A_NN transfection_NN were_VBD used_VBN to_TO model_VB changes_NNS in_IN susceptibility_NN to_TO NK_NN cell_NN killing_NN caused_VBN by_IN transient_JJ vs_CC stable_JJ E1A_NN expression_NN in_IN human_JJ cells_NNS ._. 
Only_RB stably_RB transfected_VBN target_NN cells_NNS exhibited_VBD cytolytic_JJ susceptibility_NN ,_, despite_IN expression_NN of_IN equivalent_JJ levels_NNS of_IN E1A_NN proteins_NNS in_IN Ad-infected_JJ targets_NNS ._. 
The_DT inability_NN of_IN E1A_NN gene_NN products_NNS to_TO induce_VB cytolytic_JJ susceptibility_NN during_IN infection_NN was_VBD not_RB explained_VBN by_IN an_DT inhibitory_JJ effect_NN of_IN viral_JJ infection_NN on_IN otherwise_RB susceptible_JJ target_NN cells_NNS or_CC by_IN viral_JJ gene_NN effects_NNS on_IN class_NN I_CD MHC_NN antigen_NN expression_NN on_IN target_NN cells_NNS ._. 
This_DT differential_JJ effect_NN of_IN E1A_NN expression_NN on_IN the_DT cytolytic_JJ phenotypes_NNS of_IN infected_JJ and_CC stably_RB transfected_VBN human_JJ cells_NNS suggests_VBZ that_IN human_JJ NK_NN cells_NNS provide_VBP an_DT effective_JJ immunologic_JJ barrier_NN against_IN the_DT in_FW vivo_FW survival_NN and_CC neoplastic_JJ progression_NN of_IN E1A-immortalized_JJ cells_NNS that_WDT may_MD emerge_VB from_IN the_DT reservoir_NN of_IN persistently_RB infected_JJ cells_NNS in_IN the_DT human_JJ host_NN ._. 
Distinct_JJ signaling_NN properties_NNS identify_VBP functionally_RB different_JJ CD4_NN epitopes_NNS ._. 
The_DT CD4_NN coreceptor_NN interacts_VBZ with_IN non-polymorphic_JJ regions_NNS of_IN major_JJ histocompatibility_NN complex_NN class_NN II_CD molecules_NNS on_IN antigen-presenting_JJ cells_NNS and_CC contributes_VBZ to_TO T_NN cell_NN activation_NN ._. 
We_PRP have_VBP investigated_VBN the_DT effect_NN of_IN CD4_NN triggering_NN on_IN T_NN cell_NN activating_NN signals_NNS in_IN a_DT lymphoma_NN model_NN using_VBG monoclonal_JJ antibodies_NNS (_( mAb_NN )_) which_WDT recognize_VBP different_JJ CD4_NN epitopes_NNS ._. 
We_PRP demonstrate_VBP that_IN CD4_NN triggering_NN delivers_VBZ signals_NNS capable_JJ of_IN activating_VBG the_DT NF-AT_NN transcription_NN factor_NN which_WDT is_VBZ required_VBN for_IN interleukin-2_NN gene_NN expression_NN ._. 
Whereas_IN different_JJ anti-CD4_JJ mAb_NN or_CC HIV-1_NN gp120_NN could_MD all_DT trigger_VB activation_NN of_IN the_DT protein_NN tyrosine_NN kinases_NNS p56lck_NN and_CC p59fyn_NN and_CC phosphorylation_NN of_IN the_DT Shc_NN adaptor_NN protein_NN ,_, which_WDT mediates_VBZ signals_NNS to_TO Ras_NN ,_, they_PRP differed_VBD significantly_RB in_IN their_PRP$ ability_NN to_TO activate_VB NF-AT_NN ._. 
Lack_NN of_IN full_JJ activation_NN of_IN NF-AT_NN could_MD be_VB correlated_VBN to_TO a_DT dramatically_RB reduced_VBN capacity_NN to_TO induce_VB calcium_NN flux_NN and_CC could_MD be_VB complemented_VBN with_IN a_DT calcium_NN ionophore_NN ._. 
The_DT results_NNS identify_VBP functionally_RB distinct_JJ epitopes_NNS on_IN the_DT CD4_NN coreceptor_NN involved_VBN in_IN activation_NN of_IN the_DT Ras_* /protein_NN kinase_NN C_NN and_CC calcium_NN pathways_NNS ._. 
Ligand-dependent_JJ repression_NN of_IN the_DT erythroid_JJ transcription_NN factor_NN GATA-1_NN by_IN the_DT estrogen_NN receptor_NN ._. 
High-dose_JJ estrogen_NN administration_NN induces_VBZ anemia_NN in_IN mammals_NNS ._. 
In_IN chickens_NNS ,_, estrogens_NNS stimulate_VBP outgrowth_NN of_IN bone_NN marrow-derived_JJ erythroid_JJ progenitor_NN cells_NNS and_CC delay_VBP their_PRP$ maturation_NN ._. 
This_DT delay_NN is_VBZ associated_VBN with_IN down-regulation_NN of_IN many_JJ erythroid_JJ cell-specific_JJ genes_NNS ,_, including_VBG alpha-_NN and_CC beta-_* globin_NN ,_, band_NN 3_CD ,_, band_NN 4.1_CD ,_, and_CC the_DT erythroid_JJ cell-specific_JJ histone_NN H5_NN ._. 
We_PRP show_VBP here_RB that_IN estrogens_NNS also_RB reduce_VBP the_DT number_NN of_IN erythroid_JJ progenitor_NN cells_NNS in_IN primary_JJ human_JJ bone_NN marrow_NN cultures_NNS ._. 
To_TO address_VB potential_JJ mechanisms_NNS by_IN which_WDT estrogens_NNS suppress_VBP erythropoiesis_NN ,_, we_PRP have_VBP examined_VBN their_PRP$ effects_NNS on_IN GATA-1_NN ,_, an_DT erythroid_JJ transcription_NN factor_NN that_WDT participates_VBZ in_IN the_DT regulation_NN of_IN the_DT majority_NN of_IN erythroid_JJ cell-specific_JJ genes_NNS and_CC is_VBZ necessary_JJ for_IN full_JJ maturation_NN of_IN erythrocytes_NNS ._. 
We_PRP demonstrate_VBP that_IN the_DT transcriptional_JJ activity_NN of_IN GATA-1_NN is_VBZ strongly_RB repressed_VBN by_IN the_DT estrogen_NN receptor_NN (_( ER_NN )_) in_IN a_DT ligand-dependent_JJ manner_NN and_CC that_IN this_DT repression_NN is_VBZ reversible_JJ in_IN the_DT presence_NN of_IN 4-hydroxytamoxifen_NN ._. 
ER_* -mediated_JJ repression_NN of_IN GATA-1_NN activity_NN occurs_VBZ on_IN an_DT artificial_JJ promoter_NN containing_VBG a_DT single_JJ GATA-binding_JJ site_NN ,_, as_RB well_RB as_IN in_IN the_DT context_NN of_IN an_DT intact_JJ promoter_NN which_WDT is_VBZ normally_RB regulated_VBN by_IN GATA-1_NN ._. 
GATA-1_NN and_CC ER_NN bind_VBP to_TO each_DT other_JJ in_FW vitro_FW in_IN the_DT absence_NN of_IN DNA_NN ._. 
In_IN coimmunoprecipitation_NN experiments_NNS using_VBG transfected_VBN COS_NN cells_NNS ,_, GATA-1_NN and_CC ER_NN associate_VBP in_IN a_DT ligand-dependent_JJ manner_NN ._. 
Mapping_NN experiments_NNS indicate_VBP that_IN GATA-1_NN and_CC the_DT ER_NN form_VBP at_IN least_JJS two_CD contacts_NNS ,_, which_WDT involve_VBP the_DT finger_NN region_NN and_CC the_DT N-terminal_JJ activation_NN domain_NN of_IN GATA-1_NN ._. 
We_PRP speculate_VBP that_IN estrogens_NNS exert_VBP effects_NNS on_IN erythropoiesis_NN by_IN modulating_VBG GATA-1_NN activity_NN through_IN protein-protein_JJ interaction_NN with_IN the_DT ER_NN ._. 
(_( ABSTRACT_NN TRUNCATED_VBN AT_IN 250_CD WORDS_NNS )_) 
Mouse_NN interleukin-2_NN receptor_NN alpha_NN gene_NN expression_NN ._. 
Interleukin-1_NN and_CC interleukin-2_NN control_VBP transcription_NN via_IN distinct_JJ cis-acting_JJ elements_NNS ._. 
We_PRP have_VBP shown_VBN that_IN interleukin-1_NN (_( IL-1_NN )_) and_CC IL-2_NN control_VBP IL-2_NN receptor_NN alpha_NN (_( IL-2R_NN alpha_NN )_) gene_NN transcription_NN in_IN T_NN lymphocyte_NN precursors_NNS ._. 
Here_RB we_PRP map_VBP the_DT cis-acting_JJ elements_NNS that_WDT mediate_VBP interleukin_NN responsiveness_NN of_IN the_DT mouse_NN IL-2R_NN alpha_NN gene_NN using_VBG a_DT thymic_JJ lymphoma-derived_JJ hybridoma_NN (_( PC60_NN )_) ._. 
The_DT transcriptional_JJ response_NN of_IN the_DT IL-2R_NN alpha_NN gene_NN to_TO stimulation_NN by_IN IL-1_NN +_CC IL-2_NN is_VBZ biphasic_JJ ._. 
IL-1_NN induces_VBZ a_DT rapid_JJ ,_, protein_NN synthesis-independent_JJ appearance_NN of_IN IL-2R_NN alpha_NN mRNA_NN that_WDT is_VBZ blocked_VBN by_IN inhibitors_NNS of_IN NF-kappa_NN B_NN activation_NN ._. 
It_PRP also_RB primes_VBZ cells_NNS to_TO become_VB IL-2_NN responsive_JJ and_CC thereby_RB prepares_VBZ the_DT second_JJ phase_NN ,_, in_IN which_WDT IL-2_NN induces_VBZ a_DT 100-fold_RB further_JJ increase_NN in_IN IL-2R_NN alpha_NN transcripts_NNS ._. 
Transient_JJ transfection_NN experiments_NNS show_VBP that_IN several_JJ elements_NNS in_IN the_DT promoter-proximal_JJ region_NN of_IN the_DT IL-2R_NN alpha_NN gene_NN contribute_VBP to_TO IL-1_NN responsiveness_NN ,_, most_RBS importantly_RB an_DT NF-kappa_NN B_NN site_NN conserved_VBN in_IN the_DT human_JJ and_CC mouse_NN gene_NN ._. 
IL-2_NN responsiveness_NN ,_, on_IN the_DT other_JJ hand_NN ,_, depends_VBZ on_IN a_DT 78-nucleotide_JJ segment_NN 1.3_CD kilobases_NNS upstream_RB of_IN the_DT major_JJ transcription_NN start_NN site_NN ._. 
This_DT segment_NN functions_VBZ as_IN an_DT IL-2-inducible_JJ enhancer_NN and_CC lies_VBZ within_IN a_DT region_NN that_WDT becomes_VBZ DNase_NN I_CD hypersensitive_JJ in_IN normal_JJ T_NN cells_NNS in_IN which_WDT IL-2R_NN alpha_NN expression_NN has_VBZ been_VBN induced_VBN ._. 
IL-2_NN responsiveness_NN requires_VBZ three_CD distinct_JJ elements_NNS within_IN the_DT enhancer_NN ._. 
Two_CD of_IN these_DT are_VBP potential_JJ binding_VBG sites_NNS for_IN STAT_NN proteins_NNS ._. 
Hematopoietic_JJ lineage_NN commitment_NN :_: role_NN of_IN transcription_NN factors_NNS ._. 
This_DT review_NN focuses_VBZ on_IN the_DT roles_NNS of_IN transcription_NN factors_NNS in_IN hematopoietic_JJ lineage_NN commitment_NN ._. 
A_DT brief_JJ introduction_NN to_TO lineage_NN commitment_NN and_CC asymmetric_JJ cell_NN division_NN is_VBZ followed_VBN by_IN a_DT discussion_NN of_IN several_JJ methods_NNS used_VBN to_TO identify_VB transcription_NN factors_NNS important_JJ in_IN specifying_VBG hematopoietic_JJ cell_NN types_NNS ._. 
Next_RB is_VBZ presented_VBN a_DT discussion_NN of_IN the_DT use_NN of_IN embryonic_JJ stem_NN cells_NNS in_IN the_DT analysis_NN of_IN hematopoietic_JJ gene_NN expression_NN and_CC the_DT use_NN of_IN targeted_VBN gene_NN disruption_NN to_TO analyze_VB the_DT role_NN of_IN transcription_NN factors_NNS in_IN hematopoiesis_NN ._. 
Finally_RB ,_, the_DT status_NN of_IN our_PRP$ current_JJ knowledge_NN concerning_VBG the_DT roles_NNS of_IN transcription_NN factors_NNS in_IN the_DT commitment_NN to_TO erythroid_JJ ,_, myeloid_JJ and_CC lymphoid_JJ cell_NN types_NNS is_VBZ summarized_VBN ._. 
Epstein-Barr_JJ virus_NN replicative_JJ gene_NN transcription_NN during_IN de_FW novo_FW infection_NN of_IN human_JJ thymocytes_NNS :_: simultaneous_JJ early_JJ expression_NN of_IN BZLF-1_NN and_CC its_PRP$ repressor_NN RAZ_NN ._. 
Epstein-Barr_JJ virus_NN (_( EBV_NN )_) is_VBZ known_VBN to_TO infect_VB B_NN cells_NNS and_CC epithelial_JJ cells_NNS ._. 
We_PRP and_CC others_NNS have_VBP shown_VBN that_IN EBV_NN can_MD also_RB infect_VB a_DT subset_NN of_IN thymocytes_NNS ._. 
Infection_NN of_IN thymocytes_NNS was_VBD accompanied_VBN by_IN the_DT appearance_NN of_IN linear_JJ EBV_NN genome_NN within_IN 8_CD hr_NN of_IN infection_NN ._. 
Circularization_NN of_IN the_DT EBV_NN genome_NN was_VBD not_RB detected_VBN ._. 
This_DT is_VBZ in_IN contrast_NN to_TO the_DT infection_NN in_IN B_NN cells_NNS where_WRB the_DT genome_NN can_MD circularize_VB within_IN 24_CD hr_NN of_IN infection_NN ._. 
The_DT appearance_NN of_IN the_DT BamHI_NN ZLF-1_NN gene_NN product_NN ,_, ZEBRA_NN ,_, by_IN RT-PCR_NN ,_, was_VBD observed_VBN within_IN 8_CD hr_NN of_IN infection_NN ._. 
The_DT appearance_NN of_IN a_DT novel_JJ fusion_NN transcript_NN (_( RAZ_NN )_) ,_, which_WDT comprised_VBD regions_NNS of_IN the_DT BZLF-1_NN locus_NN and_CC the_DT adjacent_JJ BRLF-1_NN locus_NN ,_, was_VBD detected_VBN by_IN RT-PCR_NN ._. 
ZEBRA_NN protein_NN was_VBD also_RB identified_VBN in_IN infected_JJ thymocytes_NNS by_IN immunoprecipitation_NN ._. 
In_IN addition_NN ,_, we_PRP demonstrated_VBD that_IN the_DT EBNA-1_NN gene_NN in_IN infected_JJ thymocytes_NNS was_VBD transcribed_VBN from_IN the_DT Fp_NN promoter_NN ,_, rather_RB than_IN from_IN the_DT Cp/Wp_NN promoter_NN which_WDT is_VBZ used_VBN in_IN latently_RB infected_JJ B_NN cells_NNS ._. 
Transcripts_NNS encoding_VBG gp350/220_NN ,_, the_DT major_JJ coat_NN protein_NN of_IN EBV_NN ,_, were_VBD identified_VBN ,_, but_CC we_PRP did_VBD not_RB find_VB any_DT evidence_NN of_IN transcription_NN from_IN the_DT LMP-2A_NN or_CC EBER-1_NN loci_NNS in_IN infected_JJ thymocytes_NNS ._. 
These_DT observations_NNS suggest_VBP that_IN de_FW novo_FW EBV_NN infection_NN of_IN thymocytes_NNS differs_VBZ from_IN infection_NN of_IN B_NN cells_NNS ._. 
The_DT main_JJ difference_NN is_VBZ that_IN with_IN thymocytes_NNS ,_, no_DT evidence_NN could_MD be_VB found_VBN that_IN the_DT virus_NN ever_RB circularizes_VBZ ._. 
Rather_RB ,_, EBV_NN remains_VBZ in_IN a_DT linear_JJ configuration_NN from_IN which_WDT replicative_JJ genes_NNS are_VBP transcribed_VBN ._. 
Identification_NN and_CC purification_NN of_IN human_JJ Stat_NN proteins_NNS activated_VBN in_IN response_NN to_TO interleukin-2_NN ._. 
A_DT key_JJ cytokine_NN induced_VBN during_IN the_DT immune_JJ response_NN is_VBZ IL-2_NN ._. 
Following_VBG T_NN cell_NN activation_NN ,_, the_DT genes_NNS encoding_VBG IL-2_NN and_CC the_DT various_JJ chains_NNS of_IN its_PRP$ receptor_NN are_VBP transcriptionally_RB induced_VBN ._. 
In_IN turn_NN ,_, secreted_VBN IL-2_NN serves_VBZ to_TO stimulate_VB the_DT proliferation_NN and_CC differentiation_NN of_IN T_NN lymphocytes_NNS ._. 
Several_JJ recent_JJ studies_NNS have_VBP implicated_VBN Jak_NN kinases_NNS in_IN the_DT signaling_NN pathway_NN induced_VBN by_IN IL-2_NN ._. 
Following_VBG this_DT lead_NN ,_, we_PRP set_VBD out_RP to_TO identify_VB transcription_NN factors_NNS induced_VBN in_IN response_NN to_TO IL-2_NN ._. 
Human_JJ peripheral_JJ blood_NN lymphocytes_NNS were_VBD observed_VBN to_TO contain_VB several_JJ IL-2_* -inducible_JJ DNA_NN binding_NN activities_NNS ._. 
Similar_JJ activities_NNS were_VBD also_RB observed_VBN in_IN a_DT transformed_VBN human_JJ lymphocyte_NN line_NN ,_, termed_VBN YT_NN ._. 
We_PRP have_VBP purified_VBN these_DT activities_NNS and_CC found_VBD that_IN the_DT principal_JJ IL-2_* -inducible_JJ component_NN bears_VBZ significant_JJ relatedness_NN to_TO a_DT prolactin-induced_JJ transcription_NN factor_NN first_RB identified_VBN in_IN sheep_NN mammary_JJ gland_NN tissue_NN ._. 
We_PRP hypothesize_VBP that_IN activation_NN of_IN this_DT protein_NN ,_, designated_VBN hStat5_NN ,_, helps_VBZ govern_VB the_DT biological_JJ effects_NNS of_IN IL-2_NN during_IN the_DT immune_JJ response_NN ._. 
E2F-1_NN and_CC a_DT cyclin-like_JJ DNA_NN repair_NN enzyme_NN ,_, uracil-DNA_NN glycosylase_NN ,_, provide_VBP evidence_NN for_IN an_DT autoregulatory_JJ mechanism_NN for_IN transcription_NN ._. 
The_DT cell_NN cycle-dependent_JJ transcription_NN factor_NN ,_, E2F-1_NN ,_, regulates_VBZ the_DT cyclin_* -like_JJ species_NNS of_IN the_DT DNA_NN repair_NN enzyme_NN uracil-DNA_NN glycosylase_NN (_( UDG_NN )_) gene_NN in_IN human_JJ osteosarcoma_NN (_( Saos-2_NN )_) cells_NNS ._. 
We_PRP demonstrate_VBP ,_, through_IN the_DT deletion_NN of_IN the_DT human_JJ UDG_NN promoter_NN sequences_NNS ,_, that_IN expression_NN of_IN E2F-1_NN activates_VBZ the_DT UDG_NN promoter_NN through_IN several_JJ E2F_NN sites_NNS ._. 
The_DT major_JJ putative_JJ downstream_JJ site_NN for_IN E2F_NN ,_, located_JJ in_IN the_DT first_JJ exon_NN ,_, serves_VBZ as_IN a_DT target_NN for_IN E2F-1/DP1_NN complex_NN binding_NN in_FW vitro_FW ._. 
We_PRP also_RB provide_VBP evidence_NN for_IN the_DT functional_JJ relationship_NN between_IN the_DT cyclin-like_JJ UDG_NN gene_NN product_NN and_CC E2F_NN ._. 
High_JJ levels_NNS of_IN UDG_NN expression_NN in_IN a_DT transient_JJ transfection_NN assay_NN result_VBP in_IN the_DT down-regulation_NN of_IN transcriptional_JJ activity_NN through_IN elements_NNS specific_JJ for_IN E2F_* -mediated_JJ transcription_NN ._. 
Overexpression_NN of_IN UDG_NN in_IN Saos_NN 2_CD cells_NNS was_VBD observed_VBN to_TO delay_VB growth_NN late_RB in_IN G1_NN phase_NN and_CC transiently_RB arrest_VB these_DT cells_NNS from_IN progressing_VBG into_IN the_DT S_NN phase_NN ._. 
This_DT hypothetical_JJ model_NN integrates_VBZ one_CD mechanism_NN of_IN DNA_NN repair_NN with_IN the_DT cell_NN cycle_NN control_NN of_IN gene_NN transcription_NN ,_, likely_RB through_IN E2F_NN ._. 
This_DT implicates_VBZ E2F_NN as_IN a_DT multifunctional_JJ target_NN for_IN proteins_NNS and_CC enzymes_NNS ,_, possibly_RB ,_, responsive_JJ to_TO DNA_NN damage_NN through_IN the_DT negative_JJ effect_NN of_IN UDG_NN on_IN E2F_* -mediated_JJ transcriptional_JJ activity_NN ._. 
Cellular_JJ and_CC molecular_JJ mechanisms_NNS of_IN IFN-gamma_NN production_NN induced_VBN by_IN IL-2_NN and_CC IL-12_NN in_IN a_DT human_JJ NK_NN cell_NN line_NN ._. 
Interferon-gamma_NN (_( IFN-gamma_NN )_) is_VBZ an_DT important_JJ immunoregulatory_JJ protein_NN produced_VBN predominantly_RB by_IN T_NN cells_NNS and_CC large_JJ granular_JJ lymphocytes_NNS (_( LGL_NN )_) in_IN response_NN to_TO different_JJ extracellular_JJ signals_NNS ._. 
In_IN particular_JJ ,_, two_CD interleukins_NNS (_( ILs_NNS )_) ,_, IL-2_NN and_CC IL-12_NN ,_, have_VBP been_VBN shown_VBN to_TO be_VB potent_JJ inducers_NNS of_IN IFN-gamma_NN gene_NN expression_NN in_IN both_CC T_NN cells_NNS and_CC LGL_NN ._. 
Although_IN it_PRP has_VBZ been_VBN reported_VBN that_IN there_EX are_VBP some_DT T_NN cell_NN lines_NNS that_WDT produce_VBP IFN-gamma_NN in_IN response_NN to_TO IL-2_NN and_CC IL-12_NN stimulation_NN ,_, there_EX has_VBZ as_RB yet_RB been_VBN no_DT report_NN of_IN a_DT natural_JJ killer_NN (_( NK_NN )_) cell_NN line_NN that_WDT responds_VBZ in_IN a_DT similar_JJ manner_NN ._. 
In_IN this_DT report_NN we_PRP present_VBP evidence_NN that_IN the_DT cell_NN line_NN NK3.3_NN derived_VBN from_IN human_JJ NK_NN cells_NNS ,_, responds_VBZ to_TO both_CC IL-2_NN and_CC IL-12_NN ,_, as_IN measured_VBN by_IN increases_NNS in_IN IFN-gamma_NN and_CC granulocyte-macrophage_JJ colony-stimulating_JJ factor_NN (_( GM-CSF_NN )_) cytoplasmic_JJ mRNA_NN and_CC protein_NN expression_NN ._. 
In_IN addition_NN ,_, when_WRB used_VBN together_RB IL-2_NN and_CC IL-12_NN synergized_VBD in_IN the_DT induction_NN of_IN IFN-gamma_NN and_CC GM-CSF_NN and_CC this_DT synergy_NN was_VBD attributed_VBN to_TO an_DT increased_VBN accumulation_NN and_CC stability_NN of_IN the_DT IFN-gamma_NN and_CC GM-CSF_NN mRNAs_NNS ._. 
To_TO investigate_VB the_DT signaling_NN pathways_NNS involved_VBN in_IN the_DT gene_NN induction_NN ,_, five_CD inhibitors_NNS ,_, cyclosporin_NN A_NN (_( CsA_NN )_) ,_, transforming_VBG growth_NN factor-beta_NN ,_, cycloheximide_NN ,_, genistein_NN ,_, and_CC staurosporine_NN A_NN ,_, were_VBD used_VBN in_IN analyzing_VBG the_DT effects_NNS of_IN IL-2_NN and_CC IL-12_NN on_IN NK3.3_NN cells_NNS ._. 
The_DT results_NNS suggest_VBP that_IN activation_NN of_IN protein_NN kinase_NN C_NN ,_, but_CC not_RB new_JJ protein_NN synthesis_NN ,_, is_VBZ required_VBN for_IN IL-2_NN induction_NN of_IN IFN-gamma_NN and_CC GM-CSF_NN cytoplasmic_JJ mRNA_NN ._. 
In_IN contrast_NN ,_, IL-12_NN induction_NN of_IN IFN-gamma_NN cytoplasmic_JJ mRNA_NN appears_VBZ to_TO only_RB partially_RB depend_VB on_IN activation_NN of_IN protein_NN kinase_NN C_NN ._. 
Furthermore_RB ,_, both_CC transforming_VBG growth_NN factor-beta_NN and_CC genistein_NN ,_, a_DT tyrosine_NN kinase_NN inhibitor_NN ,_, could_MD suppress_VB IL-2_NN and_CC IL-12_NN signaling_NN but_CC CsA_NN was_VBD generally_RB inactive_JJ ._. 
It_PRP also_RB was_VBD observed_VBN that_IN suppression_NN of_IN cytokine_NN gene_NN expression_NN by_IN these_DT agents_NNS was_VBD independent_JJ of_IN the_DT inhibition_NN of_IN proliferation_NN ._. 
In_IN addition_NN ,_, IL-2_NN but_CC not_RB IL-12_NN induced_VBD nuclear_JJ factors_NNS NF-kappa_NN B_NN and_CC AP1_NN ,_, and_CC regulation_NN of_IN the_DT nuclear_JJ levels_NNS of_IN these_DT two_CD DNA_NN binding_NN protein_NN complexes_NNS is_VBZ correlated_VBN with_IN IFN-gamma_NN and_CC GM-CSF_NN gene_NN expression_NN ._. 
These_DT data_NNS indicate_VBP that_IN IL-2_NN and_CC IL-12_NN may_MD have_VB distinct_JJ signaling_NN pathways_NNS leading_VBG to_TO the_DT induction_NN of_IN IFN-gamma_NN and_CC GM-CSF_NN gene_NN expression_NN ,_, and_CC that_IN the_DT NK3.3_NN cell_NN line_NN may_MD serve_VB as_IN a_DT novel_JJ model_NN for_IN dissecting_VBG the_DT biochemical_JJ and_CC molecular_JJ events_NNS involved_VBN in_IN these_DT pathways_NNS ._. 
A_DT functional_JJ T-cell_NN receptor_NN signaling_NN pathway_NN is_VBZ required_VBN for_IN p95vav_NN activity_NN ._. 
Stimulation_NN of_IN the_DT T-cell_NN antigen_NN receptor_NN (_( TCR_NN )_) induces_VBZ activation_NN of_IN multiple_JJ tyrosine_NN kinases_NNS ,_, resulting_VBG in_IN phosphorylation_NN of_IN numerous_JJ intracellular_JJ substrates_NNS ._. 
One_CD substrate_NN is_VBZ p95vav_NN ,_, which_WDT is_VBZ expressed_VBN exclusively_RB in_IN hematopoietic_JJ and_CC trophoblast_NN cells_NNS ._. 
It_PRP contains_VBZ a_DT number_NN of_IN structural_JJ motifs_NNS ,_, including_VBG Src_NN homology_NN 2_CD ,_, Src_NN homology_NN 3_CD ,_, and_CC pleckstrin_NN homology_NN domains_NNS and_CC a_DT putative_JJ guanine_NN nucleotide_NN exchange_NN domain_NN ._. 
The_DT role_NN of_IN p95vav_NN in_IN TCR_* -mediated_JJ signaling_NN processes_NNS is_VBZ unclear_JJ ._. 
Here_RB ,_, we_PRP show_VBP that_IN overexpression_NN of_IN p95vav_NN alone_RB in_IN Jurkat_NN T_NN cells_NNS leads_VBZ to_TO activation_NN of_IN the_DT nuclear_JJ factors_NNS ,_, including_VBG NFAT_NN ,_, involved_VBN in_IN interleukin-2_NN expression_NN ._. 
Furthermore_RB ,_, p95vav_NN synergizes_VBZ with_IN TCR_NN stimulation_NN in_IN inducing_VBG NFAT-_NN and_CC interleukin-2-_* dependent_JJ transcription_NN ._. 
In_IN contrast_NN ,_, NFAT_NN activation_NN by_IN a_DT G-protein-coupled_JJ receptor_NN is_VBZ not_RB modulated_VBN by_IN p95vav_NN overexpression_NN ,_, suggesting_VBG that_IN the_DT effect_NN is_VBZ specific_JJ to_TO the_DT TCR_NN signaling_NN pathways_NNS ._. 
Although_IN removal_NN of_IN the_DT first_JJ 67_CD amino_NN acids_NNS of_IN p95vav_NN activates_VBZ its_PRP$ transforming_NN potential_NN in_IN NIH_NN 3T3_NN cells_NNS ,_, this_DT region_NN appears_VBZ to_TO be_VB required_VBN for_IN its_PRP$ function_NN in_IN T_NN cells_NNS ._. 
We_PRP further_RB demonstrate_VBP that_IN the_DT p95vav_* -induced_JJ NFAT_NN activation_NN is_VBZ not_RB mimicked_VBN by_IN Ras_NN activation_NN ,_, though_IN its_PRP$ function_NN is_VBZ dependent_JJ upon_IN Ras_NN and_CC Raf_NN ._. 
Furthermore_RB ,_, the_DT activating_NN function_NN of_IN p95vav_NN is_VBZ blocked_VBN by_IN FK506_NN ,_, suggesting_VBG that_IN its_PRP$ activity_NN also_RB depends_VBZ on_IN calcineurin_NN ._. 
To_TO further_RB dissect_VB p95vav_NN involvement_NN in_IN TCR_NN signaling_NN ,_, we_PRP analyzed_VBD various_JJ Jurkat_NN mutants_NNS deficient_JJ in_IN TCR_NN signaling_NN function_NN or_CC TCR_NN expression_NN and_CC showed_VBD that_IN an_DT intact_JJ TCR_NN signaling_NN pathway_NN is_VBZ required_VBN for_IN p95vav_NN to_TO function_VB ._. 
However_RB ,_, overexpression_NN of_IN p95vav_NN does_VBZ not_RB appear_VB to_TO influence_VB TCR_* -induced_JJ protein_NN tyrosine_NN phosphorylation_NN or_CC increases_NNS in_IN cytoplasmic_JJ free_JJ calcium_NN ._. 
Taken_VBN together_RB ,_, our_PRP$ data_NNS suggest_VBP that_IN p95vav_NN plays_VBZ an_DT important_JJ role_NN at_IN an_DT yet_RB unidentified_JJ proximal_JJ position_NN in_IN the_DT TCR_NN signaling_NN cascade_NN ._. 
Positive_JJ and_CC negative_JJ regulation_NN of_IN granulocyte-macrophage_JJ colony-stimulating_JJ factor_NN promoter_NN activity_NN by_IN AML1-related_JJ transcription_NN factor_NN ,_, PEBP2_NN ._. 
The_DT granulocyte-macrophage_JJ colony-stimulating_JJ factor_NN (_( GM-CSF_NN )_) gene_NN promoter_NN contains_VBZ a_DT consensus_NN sequence_NN for_IN the_DT polyomavirus_NN enhancer_NN binding-protein_NN 2_CD (_( PEBP2_NN )_) transcription_NN factor_NN ,_, which_WDT consists_VBZ of_IN alpha_NN and_CC beta_NN subunits_NNS ._. 
There_EX are_VBP at_IN least_JJS two_CD genes_NNS ,_, alpha_NN A_NN and_CC alpha_NN B_NN ,_, encoding_VBG the_DT alpha_NN subunit_NN ._. 
alpha_NN B_NN is_VBZ the_DT mouse_NN homologue_NN of_IN human_JJ AML1_NN gene_NN detected_VBN at_IN the_DT breakpoints_NNS of_IN t_NN (_( 8_CD ;_: 21_CD )_) and_CC t_NN (_( 3_CD ;_: 21_CD )_) myeloid_JJ leukemias_NNS ._. 
We_PRP examined_VBD alpha_NN A1_NN (_( an_DT alpha_NN A_* -gene_JJ product_NN )_) and_CC alpha_NN B1_NN and_CC alpha_NN B2_NN (_( two_CD alpha_NN B_* -encoded_JJ isomers_NNS )_) for_IN their_PRP$ effects_NNS on_IN the_DT GM-CSF_NN promoter_NN ._. 
PEBP2_NN alpha_NN A1_NN ,_, alpha_NN B1_NN ,_, and_CC alpha_NN B2_NN proteins_NNS bound_VBD the_DT PEBP2_NN site_NN within_IN the_DT mouse_NN GM-CSF_NN promoter_NN ._. 
PEBP2_NN alpha_NN A1_NN and_CC alpha_NN B1_NN enhanced_VBD the_DT expression_NN of_IN the_DT GM-CSF_NN promoter-driven_JJ reporter_NN plasmid_NN in_IN unstimulated_JJ and_CC 12-O-tetradecanoylphorbol_NN 13-acetate/phytohemagglutinin-stimulated_JJ human_JJ Jurkat_NN T_NN cells_NNS ._. 
In_IN contrast_NN ,_, the_DT promoter_NN activity_NN was_VBD suppressed_VBN by_IN alpha_NN B2_NN ._. 
Coexpression_NN of_IN alpha_NN B1_NN and_CC alpha_NN B2_NN showed_VBD that_IN the_DT promoter_NN activity_NN could_MD be_VB determined_VBN by_IN the_DT alpha_NN B1_* /alpha_NN B2_NN ratio_NN ._. 
Jurkat_NN cell_NN extract_NN contained_VBD PEBP2_NN site-binding_JJ protein_NN (_( s_NNS )_) that_WDT cross-reacted_VBD with_IN antimouse_JJ alpha_NN A1_NN antibodies_NNS ._. 
Northern_NN blot_NN analysis_NN indicated_VBD the_DT expression_NN of_IN human_JJ PEBP2_NN alpha_NN A_NN ,_, alpha_NN B_NN (_( AML1_NN )_) ,_, and_CC beta_NN genes_NNS in_IN Jurkat_NN cells_NNS ._. 
Although_IN further_JJ studies_NNS are_VBP required_VBN to_TO determine_VB the_DT precise_JJ role_NN of_IN PEBP2_NN in_IN the_DT GM-CSF_NN promoter_NN activity_NN ,_, the_DT present_JJ findings_NNS suggested_VBD the_DT importance_NN of_IN the_DT relative_JJ ratio_NN of_IN different_JJ PEBP2_NN isoforms_NNS in_IN regulating_VBG the_DT levels_NNS of_IN the_DT promoter_NN activity_NN ._. 
IFN-gamma_NN priming_NN of_IN monocytes_NNS enhances_VBZ LPS_* -induced_JJ TNF_NN production_NN by_IN augmenting_VBG both_CC transcription_NN and_CC MRNA_NN stability_NN ._. 
The_DT induction_NN of_IN cytokine_NN expression_NN in_IN monocytes_NNS /_* macrophages_NNS by_IN bacterial_JJ endotoxin_NN or_CC lipopolysaccharide_NN is_VBZ a_DT critical_JJ ,_, highly_RB regulated_VBN host_NN defence_NN response_NN ._. 
The_DT augmentation_NN of_IN LPS_NN responses_NNS by_IN interferon_NN gamma_NN (_( IFN-gamma_NN )_) ,_, referred_VBN to_TO as_IN priming_NN ,_, is_VBZ well_RB established_VBN ._. 
However_RB ,_, the_DT mechanism_NN (_( s_NNS )_) by_IN which_WDT priming_NN occurs_VBZ is_VBZ poorly_RB defined_VBN ._. 
Using_VBG tumour_NN necrosis_NN factor_NN (_( TNF_NN )_) induction_NN as_IN a_DT model_NN ,_, experiments_NNS were_VBD designed_VBN to_TO analyse_VB in_IN detail_NN the_DT priming_NN effect_NN on_IN the_DT LPS_NN response_NN in_IN human_JJ monocytes_NNS ._. 
Priming_NN by_IN IFN-gamma_NN was_VBD primarily_RB manifested_VBN at_IN the_DT level_NN of_IN TNF_NN mRNA_NN accumulation_NN ._. 
IFN-gamma_NN pre-treatment_NN affected_VBD the_DT magnitude_NN rather_RB than_IN the_DT sensitivity_NN of_IN the_DT LPS_NN response_NN ._. 
Priming_NN occurred_VBD after_IN several_JJ hours_NNS of_IN treatment_NN ,_, and_CC the_DT primed_JJ state_NN was_VBD induced_VBN by_IN either_CC IFN-gamma_NN or_CC GM-CSF_NN ,_, but_CC not_RB M-CSF_NN ._. 
Primed_VBN monocytes_NNS transcribed_VBD TNF_NN mRNA_NN at_IN a_DT higher_JJR rate_NN than_IN freshly_RB isolated_VBN monocytes_NNS upon_IN activation_NN with_IN LPS_NN ._. 
The_DT increased_VBN transcriptional_JJ rate_NN correlated_VBD with_IN a_DT marked_JJ increase_NN in_IN nuclear_JJ factor-kappa_NN B_NN activity_NN in_IN these_DT cells_NNS as_IN determined_VBN by_IN electrophoretic_JJ mobility_NN shift_NN assay_NN using_VBG a_DT consensus_NN NF-kappa_NN B_NN oligonucleotide_NN ._. 
An_DT additional_JJ significant_JJ finding_NN was_VBD than_IN TNF_NN mRNA_NN induced_VBN in_IN primed_VBN cells_NNS was_VBD much_RB more_RBR stable_JJ than_IN in_IN unprimed_JJ cells_NNS (_( T1/2_NN increased_VBN 6-8-fold_RB )_) ._. 
Consistent_JJ with_IN the_DT increased_VBN mRNA_NN stability_NN ,_, the_DT duration_NN of_IN mRNA_NN accumulation_NN was_VBD longer_RBR following_VBG LPS_NN stimulation_NN in_IN primed_VBN monocytes_NNS ,_, in_IN addition_NN to_TO being_VBG of_IN greater_JJR magnitude_NN ._. 
Finally_RB ,_, primed_VBN and_CC unprimed_JJ cells_NNS possessed_VBD a_DT differential_JJ sensitivity_NN to_TO the_DT kinase_NN inhibitor_NN H-89_NN ._. 
H-89_NN substantially_RB suppressed_VBD LPS_* -induced_JJ TNF_NN mRNA_NN accumulation_NN in_IN unprimed_JJ cells_NNS ,_, but_CC had_VBD no_DT effect_NN on_IN primed_VBN monocytes_NNS following_VBG LPS_NN stimulation_NN ._. 
(_( ABSTRACT_NN TRUNCATED_VBN AT_IN 250_CD WORDS_NNS )_) 
A_DT Myc-associated_JJ zinc_NN finger_NN protein_NN binding_NN site_NN is_VBZ one_CD of_IN four_CD important_JJ functional_JJ regions_NNS in_IN the_DT CD4_NN promoter_NN ._. 
The_DT CD4_NN promoter_NN plays_VBZ an_DT important_JJ role_NN in_IN the_DT developmental_JJ control_NN of_IN CD4_NN transcription_NN ._. 
In_IN this_DT report_NN ,_, we_PRP show_VBP that_IN the_DT minimal_JJ CD4_NN promoter_NN has_VBZ four_CD factor_NN binding_NN sites_NNS ,_, each_DT of_IN which_WDT is_VBZ required_VBN for_IN full_JJ function_NN ._. 
Using_VBG biochemical_JJ and_CC mutagenesis_NN analyses_NNS ,_, we_PRP determined_VBD that_IN multiple_JJ nuclear_JJ factors_NNS bind_VBP to_TO these_DT independent_JJ sites_NNS ._. 
We_PRP determined_VBD that_IN an_DT initiator-like_JJ sequence_NN present_JJ at_IN the_DT cap_NN site_NN and_CC an_DT Ets_NN consensus_NN sequence_NN are_VBP required_VBN for_IN full_JJ promoter_NN function_NN ._. 
We_PRP also_RB demonstrate_VBP that_IN the_DT Myc-associated_JJ zinc_NN finger_NN protein_NN (_( MAZ_NN )_) appears_VBZ to_TO be_VB the_DT predominant_JJ factor_NN binding_VBG to_TO one_CD of_IN these_DT sites_NNS ._. 
This_DT last_JJ site_NN closely_RB resembles_VBZ the_DT ME1a1_NN G3AG4AG3_NN motif_NN previously_RB shown_VBN to_TO be_VB a_DT critical_JJ element_NN in_IN the_DT P2_NN promoter_NN of_IN the_DT c-myc_NN gene_NN ._. 
We_PRP therefore_RB believe_VBP that_IN the_DT MAZ_NN transcription_NN factor_NN is_VBZ also_RB likely_JJ to_TO play_VB an_DT important_JJ role_NN in_IN the_DT control_NN of_IN developmental_JJ expression_NN of_IN the_DT CD4_NN gene_NN ._. 
Erythropoietin_NN stimulates_VBZ transcription_NN of_IN the_DT TAL1/SCL_NN gene_NN and_CC phosphorylation_NN of_IN its_PRP$ protein_NN products_NNS ._. 
Activation_NN of_IN the_DT TAL1_NN (_( or_CC SCL_NN )_) gene_NN ,_, originally_RB identified_VBN through_IN its_PRP$ involvement_NN by_IN a_DT recurrent_JJ chromosomal_JJ translocation_NN ,_, is_VBZ the_DT most_RBS frequent_JJ molecular_JJ lesion_NN recognized_VBN in_IN T-cell_NN acute_JJ lymphoblastic_JJ leukemia_NN ._. 
The_DT protein_NN products_NNS of_IN this_DT gene_NN contain_VBP the_DT basic-helix-loop-helix_JJ motif_NN characteristic_JJ of_IN a_DT large_JJ family_NN of_IN transcription_NN factors_NNS that_WDT bind_VBP to_TO the_DT canonical_JJ DNA_NN sequence_NN CANNTG_NN as_IN protein_NN heterodimers_NNS ._. 
TAL1_NN expression_NN by_IN erythroid_JJ cells_NNS in_FW vivo_FW and_CC in_IN chemical-induced_JJ erythroleukemia_NN cell_NN lines_NNS in_FW vivo_FW suggested_VBD the_DT gene_NN might_MD regulate_VB aspects_NNS of_IN erythroid_JJ differentiation_NN ._. 
Since_IN the_DT terminal_JJ events_NNS of_IN erythropoiesis_NN are_VBP controlled_VBN by_IN the_DT glycoprotein_NN hormone_NN erythropoietin_NN (_( Epo_NN )_) ,_, we_PRP investigated_VBD whether_IN the_DT expression_NN or_CC activity_NN of_IN the_DT TAL1_NN gene_NN and_CC its_PRP$ protein_NN products_NNS were_VBD affected_VBN by_IN Epo_NN in_IN splenic_JJ erythroblasts_NNS from_IN mice_NNS infected_VBN with_IN an_DT anemia-inducing_JJ strain_NN of_IN Friend_NN virus_NN (_( FVA_NN cells_NNS )_) ._. 
Epo_NN elicited_VBD a_DT rapid_JJ ,_, dose-related_JJ increase_NN in_IN TAL1_NN mRNA_NN by_IN increasing_VBG transcription_NN of_IN the_DT gene_NN and_CC stabilizing_VBG one_CD of_IN its_PRP$ mRNAs_NNS ._. 
An_DT Epo_* -inducible_JJ TAL1_NN DNA_NN binding_NN activity_NN was_VBD identified_VBN in_IN FVA_NN cell_NN nuclear_JJ extracts_NNS that_WDT subsequently_RB decayed_VBD despite_IN accumulating_VBG mRNA_NN and_CC protein_NN ._. 
Induction_NN of_IN DNA_NN binding_NN activity_NN was_VBD associated_VBN temporally_RB with_IN Epo_* -induced_JJ phosphorylation_NN of_IN nuclear_JJ TAL1_NN protein_NN ._. 
These_DT results_NNS indicate_VBP that_IN Epo_NN acts_VBZ at_IN both_CC transcriptional_JJ and_CC posttranscriptional_JJ levels_NNS on_IN the_DT TAL1_NN locus_NN in_IN Friend_NN virus_* -induced_JJ erythroblasts_NNS and_CC establish_VBP a_DT link_NN between_IN Epo_NN signaling_NN mechanisms_NNS and_CC a_DT member_NN of_IN a_DT family_NN of_IN transcription_NN factors_NNS involved_VBN in_IN the_DT differentiation_NN of_IN diverse_JJ cell_NN lineages_NNS ._. 
Nonradioactive_JJ quantification_NN of_IN glucocorticoid_NN receptor_NN expression_NN during_IN differentiation_NN of_IN human_JJ monocytic_JJ cells_NNS (_( U937_NN )_) ._. 
We_PRP describe_VBP a_DT method_NN for_IN relative_JJ quantification_NN of_IN specific_JJ mRNA_NN using_VBG a_DT nonradioactive_JJ assay_NN based_VBN on_IN DNA_NN strand_NN competition_NN between_IN identical_JJ sequences_NNS of_IN biotin-_NN and_CC fluorescein-labeled_JJ amplicon_NN (_( probe_NN )_) and_CC unlabeled_JJ amplicon_NN (_( target_NN )_) during_IN hybridization_NN ._. 
As_IN the_DT target_NN quantity_NN increased_VBD ,_, that_DT of_IN the_DT double-labeled_JJ probe_NN decreased_VBD in_IN accordance_NN with_IN the_DT mass_NN action_NN law_NN ._. 
This_DT technique_NN was_VBD successfully_RB applied_VBN to_TO evaluate_VB differences_NNS in_IN glucocorticoid_NN receptor_NN expression_NN in_IN U937_NN cells_NNS before_IN and_CC after_IN the_DT addition_NN of_IN potent_JJ differentiation_NN inducers_NNS :_: 12-O-tetradecanoylphorbol_NN 13-acetate_NN (_( TPA_NN )_) and_CC a_DT combination_NN of_IN all-trans_JJ retinoic_JJ acid_NN (_( RA_NN )_) and_CC 1,25-dihydroxyvitamin_NN D2_NN (_( VD_NN )_) ._. 
We_PRP observed_VBD that_IN TPA_NN treatment_NN was_VBD associated_VBN with_IN an_DT increase_NN in_IN specific_JJ binding_NN of_IN [3H]dexamethasone_NN and_CC up-regulation_NN of_IN GR_NN mRNA_NN while_IN no_DT enhanced_VBN GR_NN expression_NN was_VBD perceived_VBN with_IN RA_NN /VD_NN treatment_NN ._. 
Induction_NN of_IN Sp1_NN phosphorylation_NN and_CC NF-kappa_NN B_* -independent_JJ HIV_NN promoter_NN domain_NN activity_NN in_IN T_NN lymphocytes_NNS stimulated_VBN by_IN okadaic_JJ acid_NN ._. 
In_IN contrast_NN to_TO the_DT purely_RB enhancer-dependent_JJ effect_NN of_IN cytokines_NNS such_JJ as_IN TNF_NN on_IN the_DT activity_NN of_IN the_DT HIV_NN regulatory_JJ region_NN (_( LTR_NN )_) ,_, we_PRP observed_VBD that_IN okadaic_JJ acid_NN (_( OKA_NN )_) activates_VBZ HIV_NN transcription_NN through_IN both_CC the_DT enhancer_NN ,_, responding_VBG to_TO the_DT factor_NN NF-kappa_NN B_NN ,_, and_CC the_DT promoter_NN domain_NN of_IN the_DT LTR_NN ._. 
The_DT inducibility_NN of_IN HIV_NN LTR_* -driven_JJ luciferase_NN expression_NN constructs_NNS in_IN lymphoblastoid_JJ cells_NNS stimulated_VBN by_IN OKA_NN depended_VBD on_IN both_CC functional_JJ Sp1_NN binding_NN elements_NNS and_CC the_DT ability_NN of_IN the_DT TATA_NN box_NN to_TO bind_VB the_DT protein_NN TBP_NN ._. 
In_IN both_CC transformed_VBN and_CC normal_JJ lymphocytes_NNS ,_, OKA_NN stimulation_NN induced_VBD intense_JJ phosphorylation_NN of_IN the_DT constitutively_RB expressed_VBN Sp1_NN protein_NN in_IN the_DT nucleus_NN ,_, a_DT property_NN of_IN OKA_NN not_RB shared_VBN by_IN TNF_NN ,_, phorbol_NN ester_NN ,_, or_CC PHA_NN and_CC interleukin_NN 2_CD ._. 
Responsiveness_NN of_IN LTR_NN constructs_NNS deleted_VBN of_IN kappa_NN B_NN elements_NNS to_TO HIV_NN Tat_NN expression_NN was_VBD increased_VBN upon_IN OKA_NN but_CC not_RB TNF_NN stimulation_NN ._. 
Our_PRP$ results_NNS suggest_VBP that_IN SP1_NN phosphorylation_NN induced_VBN by_IN OKA_NN ,_, a_DT selective_JJ inhibitor_NN of_IN the_DT serine-threonine_JJ phosphatase_NN PP2A_NN ,_, facilitates_VBZ the_DT formation_NN of_IN a_DT transcription_NN complex_NN involving_VBG general_JJ transcription_NN factors_NNS ,_, HIV_NN Tat_NN ,_, and_CC Sp1_NN proteins_NNS ._. 
The_DT formation_NN of_IN this_DT complex_NN would_MD increase_VB ,_, independently_RB of_IN an_DT in_NN synergy_NN with_IN NF-kappa_NN B_NN ,_, the_DT low_JJ basal_JJ activity_NN of_IN the_DT HIV_NN LTR_NN observed_VBN in_IN normal_JJ T_NN lymphocytes_NNS ._. 
The_DT role_NN of_IN shared_JJ receptor_NN motifs_NNS and_CC common_JJ Stat_NN proteins_NNS in_IN the_DT generation_NN of_IN cytokine_NN pleiotropy_NN and_CC redundancy_NN by_IN IL-2_NN ,_, IL-4_NN ,_, IL-7_NN ,_, IL-13_NN ,_, and_CC IL-15_NN ._. 
To_TO understand_VB the_DT molecular_JJ bases_NNS for_IN cytokine_NN redundancy_NN and_CC pleiotropy_NN ,_, we_PRP have_VBP compared_VBN the_DT Stat_NN proteins_NNS activated_VBN in_IN peripheral_JJ blood_NN lymphocytes_NNS (_( PBLs_NNS )_) by_IN cytokines_NNS with_IN shared_JJ and_CC distinct_JJ actions_NNS ._. 
Interleukin-2_NN (_( IL-2_NN )_) rapidly_RB activated_VBD Stat5_NN in_IN fresh_JJ PBL_NN ,_, and_CC Stat3_NN and_CC Stat5_NN in_IN preactivated_JJ PBL_NN ._. 
IL-7_NN and_CC IL-15_NN induced_VBD the_DT same_JJ complexes_NNS as_IN IL-2_NN ,_, a_DT feature_NN explained_VBN by_IN the_DT existence_NN of_IN similar_JJ tyrosine_* -phosphorylated_JJ motifs_NNS in_IN the_DT cytoplasmic_JJ domains_NNS of_IN IL-2R_NN beta_NN and_CC IL-7R_NN that_WDT can_MD serve_VB as_IN docking_NN sites_NNS for_IN Stat_NN proteins_NNS ._. 
IL-13_NN Induced_VBD the_DT same_JJ complexes_NNS as_IN IL-4_NN ,_, a_DT finding_NN explained_VBN by_IN our_PRP$ studies_NNS implicating_VBG IL-4R_NN as_IN a_DT shared_JJ component_NN of_IN the_DT receptors_NNS ._. 
These_DT studies_NNS demonstrate_VBP that_IN a_DT single_JJ cytokine_NN can_MD activate_VB different_JJ combinations_NNS of_IN Stat_NN proteins_NNS under_IN different_JJ physiological_JJ conditions_NNS ,_, and_CC also_RB indicate_VBP two_CD mechanisms_NNS by_IN which_WDT distinct_JJ cytokines_NNS can_MD activate_VB the_DT same_JJ Stat_NN protein_NN ._. 
Control_NN of_IN I_NN kappa_NN B-alpha_NN proteolysis_NN by_IN site-specific_JJ ,_, signal-induced_JJ phosphorylation_NN ._. 
I_NN kappa_NN B-alpha_NN inhibits_VBZ transcription_NN factor_NN NF-kappa_NN B_NN by_IN retaining_VBG it_PRP in_IN the_DT cytoplasm_NN ._. 
Various_JJ stimuli_NNS ,_, typically_RB those_DT associated_VBN with_IN stress_NN or_CC pathogens_NNS ,_, rapidly_RB inactivate_VBP I_NN kappa_NN B-alpha_NN ._. 
This_DT liberates_VBZ NF-kappa_NN B_NN to_TO translocate_VB to_TO the_DT nucleus_NN and_CC initiate_VB transcription_NN of_IN genes_NNS important_JJ for_IN the_DT defense_NN of_IN the_DT organism_NN ._. 
Activation_NN of_IN NF-kappa_NN B_NN correlates_VBZ with_IN phosphorylation_NN of_IN I_NN kappa_NN B-alpha_NN and_CC requires_VBZ the_DT proteolysis_NN of_IN this_DT inhibitor_NN ._. 
When_WRB either_CC serine-32_NN or_CC serine-36_NN of_IN I_NN kappa_NN B-alpha_NN was_VBD mutated_VBN ,_, the_DT protein_NN did_VBD not_RB undergo_VB signal-induced_JJ phosphorylation_NN or_CC degradation_NN ,_, and_CC NF-kappa_NN B_NN could_MD not_RB be_VB activated_VBN ._. 
These_DT results_NNS suggest_VBP that_IN phosphorylation_NN at_IN one_CD or_CC both_DT of_IN these_DT residues_NNS is_VBZ critical_JJ for_IN activation_NN of_IN NF-kappa_NN B_NN ._. 
Regulation_NN of_IN transcription_NN of_IN the_DT human_JJ erythropoietin_NN receptor_NN gene_NN by_IN proteins_NNS binding_VBG to_TO GATA-1_NN and_CC Sp1_NN motifs_NNS ._. 
Erythropoietin_NN (_( Epo_NN )_) ,_, the_DT primary_JJ regulator_NN of_IN the_DT production_NN of_IN erythroid_JJ cells_NNS ,_, acts_VBZ by_IN binding_VBG to_TO a_DT cell_NN surface_NN receptor_NN (_( EpoR_NN )_) on_IN erythroid_JJ progenitors_NNS ._. 
We_PRP used_VBD deletion_NN analysis_NN and_CC transfection_NN assays_NNS with_IN reporter_NN gene_NN constructs_NNS to_TO examine_VB the_DT transcription_NN control_NN elements_NNS in_IN the_DT 5'_JJ flanking_JJ region_NN of_IN the_DT human_JJ EpoR_NN gene_NN ._. 
In_IN erythroid_JJ cells_NNS most_JJS of_IN the_DT transcription_NN activity_NN was_VBD contained_VBN in_IN a_DT 150_CD bp_NN promoter_NN fragment_NN with_IN binding_VBG sites_NNS for_IN transcription_NN factors_NNS AP2_NN ,_, Sp1_NN and_CC the_DT erythroid-specific_JJ GATA-1_NN ._. 
The_DT 150_CD bp_NN hEpoR_NN promoter_NN exhibited_VBD high_JJ and_CC low_JJ activity_NN in_IN erythroid_JJ OCIM1_NN and_CC K562_NN cells_NNS ,_, respectively_RB ,_, reflecting_VBG the_DT high_JJ and_CC low_JJ levels_NNS of_IN constitutive_JJ hEpoR_NN expression_NN ._. 
The_DT GATA-1_NN and_CC Sp1_NN binding_NN sites_NNS in_IN this_DT promoter_NN lacking_VBG a_DT TATA_NN sequence_NN were_VBD necessary_JJ for_IN a_DT high_JJ level_NN of_IN transcription_NN activation_NN ._. 
Protein-DNA_JJ binding_NN studies_NNS suggested_VBD that_IN Sp1_NN and_CC two_CD other_JJ CCGCCC_NN binding_NN proteins_NNS from_IN erythroid_JJ and_CC non-erythroid_JJ cells_NNS could_MD bind_VB to_TO the_DT Sp1_NN binding_NN motif_NN ._. 
By_IN increasing_VBG GATA-1_NN levels_NNS via_IN co-transfection_NN ,_, we_PRP were_VBD able_JJ to_TO transactivate_VB the_DT hEpoR_NN promoter_NN in_IN K562_NN cells_NNS and_CC non-erythroid_JJ cells_NNS ,_, but_CC not_RB in_IN the_DT highly_RB active_JJ OCIM1_NN cells_NNS ,_, although_IN GATA-1_NN mRNA_NN levels_NNS were_VBD comparable_JJ in_IN OCIM1_NN and_CC K562_NN ._. 
Interestingly_RB ,_, when_WRB we_PRP mutated_VBD the_DT Sp1_NN site_NN ,_, resulting_VBG in_IN a_DT marked_JJ decrease_NN in_IN hEpoR_NN promoter_NN activity_NN ,_, we_PRP could_MD restore_VB transactivation_NN by_IN increasing_VBG GATA-1_NN levels_NNS in_IN OCIM1_NN cells_NNS ._. 
These_DT data_NNS suggest_VBP that_IN while_IN GATA-1_NN can_MD transactivate_VB the_DT EpoR_NN promoter_NN ,_, the_DT level_NN of_IN hEpoR_NN gene_NN expression_NN does_VBZ not_RB depend_VB on_IN GATA-1_NN alone_RB ._. 
Rather_RB ,_, hEpoR_NN transcription_NN activity_NN depends_VBZ on_IN coordination_NN between_IN Sp1_NN and_CC GATA-1_NN with_IN other_JJ cell-specific_JJ factors_NNS ,_, including_VBG possibly_RB other_JJ Sp1_* -like_JJ binding_NN proteins_NNS ,_, to_TO provide_VB high_JJ level_NN ,_, tissue-specific_JJ expression_NN ._. 
Overexpression_NN of_IN DR-nm23_NN ,_, a_DT protein_NN encoded_VBN by_IN a_DT member_NN of_IN the_DT nm23_NN gene_NN family_NN ,_, inhibits_VBZ granulocyte_NN differentiation_NN and_CC induces_VBZ apoptosis_NN in_IN 32Dc13_NN myeloid_JJ cells_NNS ._. 
Chronic_JJ myelogenous_JJ leukemia_NN evolves_VBZ in_IN two_CD clinically_RB distinct_JJ stages_NNS :_: a_DT chronic_JJ and_CC a_DT blast_NN crisis_NN phase_NN ._. 
The_DT molecular_JJ changes_NNS associated_VBN with_IN chronic_JJ phase_NN to_TO blast_NN crisis_NN transition_NN are_VBP largely_RB unknown_JJ ._. 
We_PRP have_VBP identified_VBN a_DT cDNA_NN clone_NN ,_, DR-nm23_NN ,_, differentially_RB expressed_VBN in_IN a_DT blast-crisis_JJ cDNA_NN library_NN ,_, which_WDT has_VBZ approximately_RB 70_CD %_NN sequence_NN similarity_NN to_TO the_DT putative_JJ metastatic_JJ suppressor_NN genes_NNS ,_, nm23-H1_NN and_CC nm23-H2_NN ._. 
The_DT deduced_VBN amino_NN acid_NN sequence_NN similarity_NN to_TO the_DT proteins_NNS encoded_VBN by_IN these_DT two_CD latter_JJ genes_NNS is_VBZ approximately_RB 65_CD %_NN and_CC includes_VBZ domains_NNS and_CC amino_NN acid_NN residues_NNS (_( the_DT leucine_NN zipper-like_JJ and_CC the_DT RGD_NN domain_NN ,_, a_DT serine_NN and_CC a_DT histidine_NN residue_NN in_IN the_DT NH2-_NN and_CC in_IN the_DT COOH-terminal_JJ portion_NN of_IN the_DT protein_NN ,_, respectively_RB )_) postulated_VBN to_TO be_VB important_JJ for_IN nm23_NN function_NN ._. 
DR-nm23_NN mRNA_NN is_VBZ preferentially_RB expressed_VBN at_IN early_JJ stages_NNS of_IN myeloid_JJ differentiation_NN of_IN highly_RB purified_VBN CD34+_JJ cells_NNS ._. 
Its_PRP$ constitutive_JJ expression_NN in_IN the_DT myeloid_JJ precursor_NN 32Dc13_NN cell_NN line_NN ,_, which_WDT is_VBZ growth-factor_NN dependent_JJ for_IN both_CC proliferation_NN and_CC differentiation_NN ,_, results_VBZ in_IN inhibition_NN of_IN granulocytic_JJ differentiation_NN induced_VBN by_IN granulocyte_NN colony-stimulating_JJ factor_NN and_CC causes_VBZ apoptotic_JJ cell_NN death_NN ._. 
These_DT results_NNS are_VBP consistent_JJ with_IN a_DT role_NN for_IN DR-nm23_NN in_IN normal_JJ hematopoiesis_NN and_CC raise_VBP the_DT possibility_NN that_IN its_PRP$ overexpression_NN contributes_VBZ to_TO differentiation_NN arrest_NN ,_, a_DT feature_NN of_IN blastic_JJ transformation_NN in_IN chronic_JJ myelogenous_JJ leukemia_NN ._. 
An_DT interferon-gamma_NN activation_NN sequence_NN mediates_VBZ the_DT transcriptional_JJ regulation_NN of_IN the_DT IgG_NN Fc_NN receptor_NN type_NN IC_NN gene_NN by_IN interferon-gamma_NN ._. 
Expression_NN of_IN the_DT IgG_NN Fc_NN receptor_NN type_NN I_CD (_( Fc_NN gamma_NN RI_NN )_) on_IN myeloid_JJ cells_NNS is_VBZ dramatically_RB increased_VBN by_IN treatment_NN with_IN interferon-gamma_NN (_( IFN-gamma_NN )_) ._. 
We_PRP observed_VBD that_IN Fc_NN gamma_NN RI_NN transcript_NN levels_NNS in_IN monoblast-like_JJ U937_NN cells_NNS were_VBD elevated_VBN within_IN 3_CD hr_NN and_CC peaked_VBD 12_CD hr_NN after_IN exposure_NN to_TO IFN-gamma_NN ._. 
Treatment_NN of_IN U937_NN with_IN IFN-gamma_NN for_IN 9_CD hr_NN in_IN the_DT presence_NN of_IN cycloheximide_NN led_VBD to_TO super-induction_NN of_IN Fc_NN gamma_NN RI_NN expression_NN ._. 
Nuclear_JJ run-on_JJ analysis_NN revealed_VBD that_IN the_DT rate_NN of_IN Fc_NN gamma_NN RI_NN transcription_NN was_VBD increased_VBN by_IN IFN-gamma_NN ._. 
Genomic_JJ sequence_NN upstream_JJ of_IN the_DT Fc_NN gamma_NN RIC_NN gene_NN was_VBD cloned_VBN and_CC subjected_VBN to_TO primer_NN extension_NN analysis_NN ,_, which_WDT demonstrated_VBD a_DT single_JJ transcription_NN initiation_NN site_NN without_IN a_DT TATA_NN box_NN ._. 
Transient_JJ transfections_NNS of_IN CAT_NN reporter_NN gene_NN constructs_NNS containing_VBG various_JJ Fc_NN gamma_NN RIC_NN promoter_NN sequences_NNS into_IN U937_NN cells_NNS revealed_VBD that_IN a_DT 20-bp_JJ region_NN surrounding_VBG the_DT transcription_NN start_NN site_NN (_( -7_CD to_TO +13_CD )_) was_VBD capable_JJ of_IN mediating_VBG transcription_NN initiation_NN and_CC that_IN an_DT IFN-gamma_NN responsive_JJ element_NN (_( GIRE_NN )_) was_VBD present_JJ within_IN 74_CD bp_NN upstream_JJ of_IN the_DT transcription_NN initiation_NN site_NN ._. 
A_DT 17-bp_JJ sequence_NN between_IN positions_NNS -51_CD and_CC -35_CD conferred_VBD IFN-gamma_NN responsiveness_NN on_IN a_DT heterologous_JJ promoter_NN ._. 
Double-stranded_JJ GIRE_NN sequence_NN ,_, but_CC not_RB a_DT scrambled_VBN sequence_NN ,_, was_VBD specifically_RB bound_VBN by_IN nuclear_JJ proteins_NNS from_IN IFN-gamma_NN treated_JJ U937_NN cells_NNS ._. 
Gel_NN shift_NN experiments_NNS further_RB showed_VBD that_IN the_DT STAT1_NN alpha_NN protein_NN bound_VBD to_TO the_DT Fc_NN gamma_NN RIC_NN GIRE_NN in_IN response_NN to_TO IFN-gamma_NN treatment_NN of_IN U937_NN cells_NNS ._. 
The_DT Fc_NN gamma_NN RIC_NN GIRE_NN is_VBZ homologous_JJ to_TO the_DT IFN-gamma_NN activation_NN sequence_NN (_( GAS_NN )_) of_IN the_DT guanylate_NN binding_NN protein_NN and_CC to_TO X_NN box_NN elements_NNS of_IN class_NN II_CD MHC_NN genes_NNS ._. 
Our_PRP$ results_NNS demonstrate_VBP that_IN transcriptional_JJ regulation_NN of_IN the_DT Fc_NN gamma_NN RIC_NN gene_NN by_IN IFN-gamma_NN involves_VBZ the_DT binding_NN of_IN STAT1_NN alpha_NN to_TO a_DT 17-bp_JJ GAS_NN homology_NN in_IN the_DT proximal_JJ promoter_NN ._. 
Constitutively_RB activated_VBN Jak_* -STAT_JJ pathway_NN in_IN T_NN cells_NNS transformed_VBN with_IN HTLV-I_NN ._. 
Human_JJ T_NN cell_NN lymphotropic_JJ virus_NN I_NN (_( HTLV-I_NN )_) is_VBZ the_DT etiological_JJ agent_NN for_IN adult_JJ T_NN cell_NN leukemia_NN and_CC tropical_JJ spastic_JJ paraparesis_NN (_( also_RB termed_VBN HTLV-I_* -associated_JJ myelopathy_NN )_) ._. 
HTLV-I_* -infected_JJ peripheral_JJ blood_NN T_NN cells_NNS exhibit_VBP an_DT initial_JJ phase_NN of_IN interleukin-2_NN (_( IL-2_NN )_) -dependent_JJ growth_NN ;_: over_IN time_NN ,_, by_IN an_DT unknown_JJ mechanism_NN ,_, the_DT cells_NNS become_VBP IL-2_* -independent_JJ ._. 
Whereas_IN the_DT Jak_NN kinases_NNS Jak1_NN and_CC Jak3_NN and_CC the_DT signal_NN transducer_NN and_CC activator_NN of_IN transcription_NN proteins_NNS Stat3_NN and_CC Stat5_NN are_VBP activated_VBN in_IN normal_JJ T_NN cells_NNS in_IN response_NN to_TO IL-2_NN ,_, this_DT signaling_NN pathway_NN was_VBD constitutively_RB activated_VBN in_IN HTLV-I_* -transformed_JJ cells_NNS ._. 
In_IN HTLV-I_* -infected_JJ cord_NN blood_NN lymphocytes_NNS ,_, the_DT transition_NN from_IN IL-2_* -dependent_JJ to_TO IL-2_* -independent_JJ growth_NN correlated_VBD with_IN the_DT acquisition_NN of_IN a_DT constitutively_RB activated_JJ Jak_* -STAT_JJ pathway_NN ,_, which_WDT suggests_VBZ that_IN this_DT pathway_NN participates_VBZ in_IN HTLV-I_* -mediated_JJ T_NN cell_NN transformation_NN ._. 
Nitric_JJ oxide_NN decreases_VBZ cytokine_* -induced_JJ endothelial_JJ activation_NN ._. 
Nitric_JJ oxide_NN selectively_RB reduces_VBZ endothelial_JJ expression_NN of_IN adhesion_NN molecules_NNS and_CC proinflammatory_JJ cytokines_NNS ._. 
To_TO test_VB the_DT hypothesis_NN that_IN nitric_JJ oxide_NN (_( NO_NN )_) limits_VBZ endothelial_JJ activation_NN ,_, we_PRP treated_VBD cytokine-stimulated_JJ human_JJ saphenous_JJ vein_NN endothelial_JJ cells_NNS with_IN several_JJ NO_NN donors_NNS and_CC assessed_VBD their_PRP$ effects_NNS on_IN the_DT inducible_JJ expression_NN of_IN vascular_JJ cell_NN adhesion_NN molecule-1_NN (_( VCAM-1_NN )_) ._. 
In_IN a_DT concentration-dependent_JJ manner_NN ,_, NO_NN inhibited_VBD interleukin_NN (_( IL_NN )_) -1_CD alpha-stimulated_JJ VCAM-1_NN expression_NN by_IN 35-55_CD %_NN as_IN determined_VBN by_IN cell_NN surface_NN enzyme_NN immunoassays_NNS and_CC flow_NN cytometry_NN ._. 
This_DT inhibition_NN was_VBD paralleled_VBN by_IN reduced_VBN monocyte_NN adhesion_NN to_TO endothelial_JJ monolayers_NNS in_IN nonstatic_JJ assays_NNS ,_, was_VBD unaffected_JJ by_IN cGMP_NN analogues_NNS ,_, and_CC was_VBD quantitatively_RB similar_JJ after_IN stimulation_NN by_IN either_CC IL-1_NN alpha_NN ,_, IL-1_NN beta_NN ,_, IL-4_NN ,_, tumor_NN necrosis_NN factor_NN (_( TNF_NN alpha_NN )_) ,_, or_CC bacterial_JJ lipopolysaccharide_NN ._. 
NO_NN also_RB decreased_VBD the_DT endothelial_JJ expression_NN of_IN other_JJ leukocyte_NN adhesion_NN molecules_NNS (_( E-selectin_NN and_CC to_TO a_DT lesser_JJR extent_NN ,_, intercellular_JJ adhesion_NN molecule-1_NN )_) and_CC secretable_JJ cytokines_NNS (_( IL-6_NN and_CC IL-8_NN )_) ._. 
Inhibition_NN of_IN endogenous_JJ NO_NN production_NN by_IN L-N-monomethyl-arginine_NN also_RB induced_VBD the_DT expression_NN of_IN VCAM-1_NN ,_, but_CC did_VBD not_RB augment_VB cytokine-induced_JJ VCAM-1_NN expression_NN ._. 
Nuclear_JJ run-on_JJ assays_NNS ,_, transfection_NN studies_NNS using_VBG various_JJ VCAM-1_NN promoter_NN reporter_NN gene_NN constructs_NNS ,_, and_CC electrophoretic_JJ mobility_NN shift_NN assays_NNS indicated_VBD that_IN NO_NN represses_VBZ VCAM-1_NN gene_NN transcription_NN ,_, in_IN part_NN ,_, by_IN inhibiting_VBG NF-kappa_NN B_NN ._. 
We_PRP propose_VBP that_IN NO_NN 's_POS ability_NN to_TO limit_VB endothelial_JJ activation_NN and_CC inhibit_VB monocyte_NN adhesion_NN may_MD contribute_VB to_TO some_DT of_IN its_PRP$ antiatherogenic_JJ and_CC antiinflammatory_JJ properties_NNS within_IN the_DT vessel_NN wall_NN ._. 
MIP1_NN alpha_NN nuclear_JJ protein_NN (_( MNP_NN )_) ,_, a_DT novel_JJ transcription_NN factor_NN expressed_VBN in_IN hematopoietic_JJ cells_NNS that_WDT is_VBZ crucial_JJ for_IN transcription_NN of_IN the_DT human_JJ MIP-1_NN alpha_NN gene_NN ._. 
Murine_JJ macrophage_NN inflammatory_JJ protein_NN 1_CD alpha_NN (_( MIP-1_NN alpha_NN )_) and_CC its_PRP$ human_JJ equivalent_NN (_( GOS19_NN ,_, LD78_NN ,_, or_CC AT464_NN )_) are_VBP members_NNS of_IN the_DT -C-C_NN family_NN of_IN low-molecular-weight_JJ chemokines_NNS ._. 
Secreted_VBN from_IN activated_VBN T_NN cells_NNS and_CC macrophages_NNS ,_, bone_NN marrow-derived_JJ MIP-1_NN alpha_* /GOS19_NN inhibits_VBZ primitive_JJ hematopoietic_JJ stem_NN cells_NNS and_CC appears_VBZ to_TO be_VB involved_VBN in_IN the_DT homeostatic_JJ control_NN of_IN stem_NN cell_NN proliferation_NN ._. 
It_PRP also_RB induces_VBZ chemotaxis_NN and_CC inflammatory_JJ responses_NNS in_IN mature_JJ cell_NN types_NNS ._. 
Therefore_RB ,_, it_PRP is_VBZ important_JJ to_TO understand_VB the_DT mechanisms_NNS which_WDT control_VBP the_DT expression_NN of_IN MIP-1_NN alpha_* /GOS19_NN ._. 
Previous_JJ work_NN has_VBZ shown_VBN that_IN in_IN Jurkat_NN T_NN cells_NNS ,_, a_DT set_NN of_IN widely_RB expressed_VBN transcription_NN factors_NNS (_( the_DT ICK-1_NN family_NN )_) affect_VBP the_DT GOS19_NN promoter_NN ._. 
One_CD member_NN ,_, ICK-1A_NN ,_, behaves_VBZ as_IN a_DT strong_JJ negative_JJ regulator_NN ._. 
In_IN this_DT communication_NN ,_, we_PRP provide_VBP evidence_NN that_IN the_DT pathway_NN of_IN induction_NN in_IN the_DT macrophage_NN cell_NN line_NN U937_NN is_VBZ different_JJ from_IN that_DT in_IN Jurkat_NN cells_NNS ._. 
Furthermore_RB ,_, we_PRP show_VBP that_IN the_DT ICK-1_NN binding_NN site_NN does_VBZ not_RB confer_VB negative_JJ regulation_NN in_IN U937_NN cells_NNS ._. 
We_PRP provide_VBP evidence_NN for_IN an_DT additional_JJ binding_VBG site_NN ,_, the_DT MIP-1_NN alpha_NN nuclear_JJ protein_NN (_( MNP_NN )_) site_NN ,_, which_WDT overlaps_VBZ the_DT ICK-1_NN site_NN ._. 
Interaction_NN of_IN nuclear_JJ extracts_NNS from_IN various_JJ cell_NN lines_NNS and_CC tissue_NN with_IN the_DT MNP_NN site_NN leads_VBZ to_TO the_DT formation_NN of_IN fast-migrating_JJ protein-DNA_JJ complexes_NNS with_IN similar_JJ but_CC distinct_JJ electrophoretic_JJ mobilities_NNS ._. 
A_DT mutation_NN of_IN the_DT MNP_NN site_NN which_WDT does_VBZ not_RB abrogate_VB ICK-1_NN binding_NN inactivates_VBZ the_DT GOS19.1_NN promoter_NN in_IN U937_NN cells_NNS and_CC reduces_VBZ its_PRP$ activity_NN by_IN fourfold_RB in_IN Jurkat_NN cells_NNS ._. 
We_PRP propose_VBP that_IN the_DT MNP_NN protein_NN (_( s_NNS )_) binding_VBG at_IN the_DT MNP_NN site_NN constitutes_VBZ a_DT novel_JJ transcription_NN factor_NN (_( s_NNS )_) expressed_VBN in_IN hematopoietic_JJ cells_NNS ._. 
The_DT effect_NN of_IN Toremifene_NN on_IN the_DT expression_NN of_IN some_DT genes_NNS in_IN human_JJ mononuclear_JJ cells_NNS ._. 
Toremifene_NN exerts_VBZ multiple_JJ and_CC varied_JJ effects_NNS on_IN the_DT gene_NN expression_NN of_IN human_JJ peripheral_JJ mononuclear_JJ cells_NNS ._. 
After_IN short-term_JJ ,_, in_FW vitro_FW exposure_NN to_TO therapeutical_JJ levels_NNS ,_, distinct_JJ changes_NNS in_IN P-glycoprotein_NN ,_, steroid_NN receptors_NNS ,_, p53_NN and_CC Bcl-2_NN expression_NN take_VBP place_NN ._. 
In_IN view_NN of_IN the_DT increasing_VBG use_NN of_IN antiestrogens_NNS in_IN cancer_NN therapy_NN and_CC prevention_NN ,_, there_EX is_VBZ obvious_JJ merit_NN in_IN long-term_JJ in_FW vivo_FW studies_NNS to_TO be_VB conducted_VBN ._. 
The_DT interleukin-5_* /receptor_NN interaction_NN activates_VBZ Lyn_NN and_CC Jak2_NN tyrosine_NN kinases_NNS and_CC propagates_VBZ signals_NNS via_IN the_DT Ras-Raf-1-MAP_NN kinase_NN and_CC the_DT Jak_* -STAT_JJ pathways_NNS in_IN eosinophils_NNS ._. 
We_PRP have_VBP shown_VBN that_IN the_DT interaction_NN of_IN interleukin_NN (_( IL_NN )_) -5_CD with_IN the_DT receptor_NN activates_VBZ Lyn_NN tyrosine_NN kinase_NN within_IN 1_CD min_NN and_CC Jak2_NN tyrosine_NN kinase_NN within_IN 1-3_CD min_NN ._. 
IL-5_NN also_RB stimulates_VBZ GTP_NN binding_VBG to_TO p21ras_NN ._. 
The_DT signal_NN is_VBZ subsequently_RB propagated_VBN through_IN the_DT activation_NN of_IN Raf-1_NN ,_, MEK_NN ,_, and_CC MAP_NN kinases_NNS as_IN shown_VBN by_IN their_PRP$ increased_VBN autophosphorylation_NN in_FW vitro_FW and_CC phosphorylation_NN in_FW situ_FW ._. 
Jak2_NN kinase_NN has_VBZ been_VBN shown_VBN to_TO phosphorylate_VB STAT_NN nuclear_JJ proteins_NNS ._. 
The_DT activation_NN of_IN STAT_NN nuclear_JJ factors_NNS was_VBD studied_VBN by_IN electrophoretic_JJ mobility_NN shift_NN assay_NN using_VBG a_DT gamma_NN activation_NN site_NN (_( GAS_NN )_) probe_NN ._. 
We_PRP found_VBD that_IN IL-5_NN induces_VBZ two_CD GAS-binding_JJ proteins_NNS in_IN eosinophils_NNS ,_, one_CD of_IN which_WDT is_VBZ STAT1_NN ._. 
We_PRP conclude_VBP that_IN IL-5_NN induced_JJ signals_NNS are_VBP propagated_VBN through_IN two_CD distinct_JJ pathways_NNS :_: (_( 1_LS )_) Lyn_NN -->_TO Ras_NN -->_TO Raf-1_NN -->_TO MEK_NN -->_TO MAP_NN kinase_NN and_CC (_( 2_LS )_) Jak2_NN -->_TO STAT1_NN ._. 
The_DT retinoblastoma_NN gene_NN product_NN negatively_RB regulates_VBZ transcriptional_JJ activation_NN mediated_VBN by_IN the_DT human_JJ cytomegalovirus_NN IE2_NN protein_NN ._. 
The_DT IE2_NN gene_NN product_NN of_IN human_JJ cytomegalovirus_NN (_( HCMV_NN )_) is_VBZ one_CD of_IN a_DT few_JJ viral_JJ regulatory_JJ proteins_NNS expressed_VBN immediately_RB upon_IN infection_NN of_IN the_DT host_NN cell_NN ._. 
It_PRP is_VBZ a_DT potent_JJ transcriptional_JJ activator_NN of_IN many_JJ viral_JJ and_CC cellular_JJ promoters_NNS ._. 
We_PRP found_VBD that_IN the_DT retinoblastoma_NN susceptibility_NN gene_NN product_NN (_( Rb_NN )_) dramatically_RB suppressed_VBD this_DT IE2_NN transactivation_NN of_IN various_JJ promoters_NNS ._. 
However_RB ,_, unlike_IN another_DT tumor_NN suppressor_NN protein_NN ,_, p53_NN ,_, Rb_NN did_VBD not_RB have_VB any_DT significant_JJ effect_NN on_IN basal_JJ levels_NNS of_IN transcription_NN ,_, suggesting_VBG that_IN Rb_NN specifically_RB interacts_VBZ with_IN IE2_NN rather_RB than_IN other_JJ cellular_JJ factors_NNS involved_VBN in_IN the_DT general_JJ transcription_NN machinery_NN ._. 
We_PRP found_VBD by_IN protein-affinity_JJ chromatography_NN that_IN Rb_NN in_IN nuclear_JJ extracts_NNS or_CC produced_VBN by_IN in_FW vitro_FW translation_NN directly_RB bound_VBD to_TO IE2_NN ._. 
Our_PRP$ results_NNS suggest_VBP that_IN Rb_NN may_MD regulate_VB the_DT life_NN cycle_NN of_IN HCMV_NN ,_, which_WDT is_VBZ endemic_JJ in_IN the_DT human_JJ population_NN ._. 
Furthermore_RB ,_, these_DT data_NNS may_MD provide_VB new_JJ insights_NNS into_IN the_DT slow_JJ rate_NN of_IN HCMV_NN DNA_NN replication_NN in_IN cells_NNS and_CC the_DT possible_JJ involvement_NN of_IN HCMV_NN in_IN tumorigenesis_NN ._. 
Expression_NN of_IN the_DT nucleoside_NN diphosphate_NN kinase_NN in_IN human_JJ skin_NN cancers_NNS :_: an_DT immunohistochemical_JJ study_NN ._. 
Expression_NN of_IN nucleoside_NN diphosphate_NN (_( NDP_NN )_) kinase_NN ,_, which_WDT is_VBZ homologous_JJ to_TO the_DT nm23_NN gene_NN product_NN in_IN a_DT variety_NN of_IN species_NNS ,_, has_VBZ been_VBN found_VBN to_TO be_VB inversely_RB associated_VBN with_IN metastatic_JJ potential_NN ._. 
However_RB ,_, the_DT relationship_NN remains_VBZ controversial_JJ according_VBG to_TO the_DT tumor_NN cell_NN types_NNS and_CC experimental_JJ system_NN ,_, with_IN conflicting_VBG results_NNS from_IN different_JJ research_NN groups_NNS ._. 
In_IN order_NN to_TO determine_VB whether_IN NDP_NN kinase_NN expression_NN serves_VBZ as_IN a_DT marker_NN for_IN metastatic_JJ potential_NN in_IN human_JJ skin_NN cancer_NN ,_, we_PRP assessed_VBD the_DT levels_NNS of_IN NDP_NN kinase_NN expression_NN in_IN 9_CD keratoacanthomas_NNS (_( KAs_NNS )_) ,_, 26_CD squamous_JJ cell_NN carcinomas_NNS (_( SCCs_NNS )_) ,_, and_CC 25_CD basal_JJ cell_NN carcinomas_NNS (_( BCCs_NNS )_) using_VBG immunohistochemistry_NN ._. 
The_DT expression_NN of_IN NDP_NN kinase_NN was_VBD intense_JJ in_IN KA_NN and_CC SCC_NN compared_VBN with_IN BCC_NN ._. 
However_RB ,_, the_DT difference_NN of_IN NDP_NN kinase_NN expression_NN between_IN KA_NN and_CC SCC_NN was_VBD not_RB statistically_RB significant_JJ ._. 
And_CC there_EX was_VBD no_DT statistically_RB significant_JJ difference_NN in_IN NDP_NN kinase_NN expression_NN between_IN SCC_NN with_IN metastasis_NN and_CC SCC_NN without_IN metastasis_NN ._. 
Our_PRP$ results_NNS contradict_VBP the_DT hypothesis_NN concerning_VBG the_DT possible_JJ role_NN of_IN nm23_NN gene_NN as_IN a_DT metastatic_JJ suppressor_NN gene_NN in_IN human_JJ skin_NN cancer_NN ._. 
The_DT mechanism_NN of_IN overexpression_NN in_IN various_JJ tumor_NN cell_NN types_NNS and_CC its_PRP$ biological_JJ significance_NN in_IN cutaneous_JJ carcinogenesis_NN remain_VB to_TO be_VB determined_VBN ._. 
RB_NN and_CC a_DT novel_JJ E2F-1_NN binding_NN protein_NN in_IN MHC_NN class_NN II_CD deficient_JJ B-cell_NN lines_NNS and_CC normal_JJ IFN-gamma_NN induction_NN of_IN the_DT class_NN IL_NN transactivator_NN CIITA_NN in_IN class_NN II_CD non-inducible_JJ RB_* -defective_JJ tumor_NN lines_NNS ._. 
The_DT major_JJ histocompatibility_NN (_( MHC_NN )_) class_NN II_CD genes_NNS encode_VBP cell_NN surface_NN proteins_NNS that_WDT bind_VBP antigenic_JJ peptide_NN for_IN presentation_NN to_TO T-cells_NNS ._. 
The_DT class_NN II_CD proteins_NNS are_VBP expressed_VBN constitutively_RB on_IN B-cells_NNS and_CC EBV-_NN transformed_JJ B-cells_NNS ,_, and_CC are_VBP inducible_JJ by_IN IFN-gamma_NN on_IN a_DT wide_JJ variety_NN of_IN cell_NN types_NNS ._. 
Retinoblastoma_NN protein_NN (_( RB_NN )_) is_VBZ a_DT tumor_NN suppressor_NN and_CC functions_VBZ as_IN a_DT transcriptional_JJ repressor_NN by_IN binding_VBG and_CC inactivating_VBG the_DT transactivator_NN E2F-I_NN ._. 
RB_* -defective_JJ tumor_NN lines_NNS are_VBP non-inducible_JJ for_IN MHC_NN class_NN II_CD by_IN IFN-gamma_NN ,_, or_CC very_RB weakly_RB inducible_JJ ,_, but_CC transfection_NN of_IN 2_CD different_JJ lines_NNS with_IN RB_NN expression_NN vectors_NNS re-establishes_VBZ or_CC substantially_RB enhances_VBZ class_NN II_CD inducibility_NN ._. 
Therefore_RB ,_, we_PRP examined_VBD the_DT RB_NN status_NN of_IN a_DT series_NN of_IN B-cell_NN mutants_NNS that_WDT are_VBP defective_JJ in_IN class_NN II_CD expression_NN ,_, generated_VBN either_CC in_FW vitro_FW or_CC derived_VBN from_IN Bare_NN Lymphocyte_NN Syndrome_NN (_( BLS_NN )_) patients_NNS ._. 
Nuclear_JJ matrix-bound_JJ RB_NN was_VBD detectable_JJ in_IN all_DT cases_NNS ,_, indicating_VBG that_IN loss_NN of_IN RB_NN is_VBZ not_RB responsible_JJ for_IN decreased_VBN class_NN II_CD expression_NN in_IN these_DT lines_NNS ._. 
A_DT second_JJ E2F-I_NN binding_NN protein_NN ,_, most_RBS likely_RB DP-I_NN ,_, was_VBD also_RB apparently_RB normal_JJ in_IN both_CC class_NN II-positive_JJ and_CC -negative_JJ B-cell_NN lines_NNS ._. 
We_PRP also_RB examined_VBD the_DT IFN-gamma_NN induction_NN of_IN CIITA_NN in_IN RB_* -defective_JJ lines_NNS ._. 
CIITA_NN is_VBZ a_DT class_NN II_CD gene_NN transactivator_NN known_VBN to_TO be_VB defective_JJ in_IN one_CD form_NN of_IN BLS_NN and_CC to_TO be_VB required_VBN for_IN the_DT induction_NN of_IN MHC_NN class_NN II_CD by_IN IFN-gamma_NN ._. 
CIITA_NN mRNA_NN is_VBZ normally_RB inducible_JJ by_IN IFN-gamma_NN in_IN class_NN II_CD non-inducible_JJ ,_, RB_* -defective_JJ lines_NNS ,_, and_CC in_IN one_CD line_NN ,_, re-expression_NN of_IN RB_NN has_VBZ no_DT effect_NN on_IN CIITA_NN mRNA_NN induction_NN levels_NNS ._. 
Thus_RB ,_, the_DT block_NN in_IN MHC_NN class_NN II_CD inducibility_NN in_IN RB_* -defective_JJ cells_NNS is_VBZ not_RB due_JJ to_TO a_DT block_NN in_IN CIITA_NN inducibility_NN ._. 
Interleukin_NN 12_CD induces_VBZ tyrosine_NN phosphorylation_NN and_CC activation_NN of_IN STAT4_NN in_IN human_JJ lymphocytes_NNS ._. 
Interleukin_NN 12_CD (_( IL-12_NN )_) is_VBZ an_DT important_JJ immunoregulatory_JJ cytokine_NN whose_WP$ receptor_NN is_VBZ a_DT member_NN of_IN the_DT hematopoietin_NN receptor_NN superfamily_NN ._. 
We_PRP have_VBP recently_RB demonstrated_VBN that_IN stimulation_NN of_IN human_JJ T_NN and_CC natural_JJ killer_NN cells_NNS with_IN IL-12_NN induces_VBZ tyrosine_NN phosphorylation_NN of_IN the_DT Janus_NN family_NN tyrosine_NN kinase_NN JAK2_NN and_CC Tyk2_NN ,_, implicating_VBG these_DT kinases_NNS in_IN the_DT immediate_JJ biochemical_JJ response_NN to_TO IL-12_NN ._. 
Recently_RB ,_, transcription_NN factors_NNS known_VBN as_IN STATs_NNS (_( signal_NN transducers_NNS and_CC activators_NNS of_IN transcription_NN )_) have_VBP been_VBN shown_VBN to_TO be_VB tyrosine_NN phosphorylated_JJ and_CC activated_VBN in_IN response_NN to_TO a_DT number_NN of_IN cytokines_NNS that_WDT bind_VBP hematopoietin_NN receptors_NNS and_CC activate_VBP JAK_NN kinases_NNS ._. 
In_IN this_DT report_NN we_PRP demonstrate_VBP that_IN IL-12_NN induces_VBZ tyrosine_NN phosphorylation_NN of_IN a_DT recently_RB identified_VBN STAT_NN family_NN member_NN ,_, STAT4_NN ,_, and_CC show_VBP that_IN STAT4_NN expression_NN is_VBZ regulated_VBN by_IN T-cell_NN activation_NN ._. 
Furthermore_RB ,_, we_PRP show_VBP that_IN IL-12_NN stimulates_VBZ formation_NN of_IN a_DT DNA-binding_JJ complex_NN that_WDT recognizes_VBZ a_DT DNA_NN sequence_NN previously_RB shown_VBN to_TO bind_VB STAT_NN proteins_NNS and_CC that_IN this_DT complex_NN contains_VBZ STAT4_NN ._. 
These_DT data_NNS ,_, and_CC the_DT recent_JJ demonstration_NN of_IN JAK_NN phosphorylation_NN by_IN IL-12_NN ,_, identify_VBP a_DT rapid_JJ signal-transduction_JJ pathway_NN likely_JJ to_TO mediate_VB IL-12_* -induced_JJ gene_NN expression_NN ._. 
Temperature-induced_JJ down-regulation_NN of_IN the_DT glucocorticoid_NN receptor_NN in_IN peripheral_JJ blood_NN mononuclear_JJ leucocyte_NN in_IN patients_NNS with_IN sepsis_NN or_CC septic_JJ shock_NN ._. 
OBJECTIVE_NN :_: Activation_NN of_IN the_DT hypothalamic-pituitary-adrenal_JJ axis_NN is_VBZ of_IN vital_JJ importance_NN during_IN critical_JJ illness_NN ._. 
We_PRP have_VBP studied_VBN the_DT adaptive_JJ mechanisms_NNS which_WDT occur_VBP at_IN the_DT level_NN of_IN the_DT glucocorticoid_NN receptor_NN in_IN glucocorticoid_NN target_NN tissues_NNS in_IN patients_NNS with_IN sepsis_NN or_CC septic_JJ shock_NN ._. 
DESIGN_NN :_: The_DT effects_NNS of_IN hypercortisolaemia_NN ,_, hyperthermia_NN and_CC cellular_JJ composition_NN on_IN number_NN of_IN glucocorticoid_NN receptors_NNS per_IN cell_NN and_CC their_PRP$ affinity_NN were_VBD evaluated_VBN ,_, both_CC in_FW vitro_FW and_CC in_FW vivo_FW ,_, in_IN peripheral_JJ blood_NN mononuclear_JJ leucocytes_NNS of_IN control_JJ subjects_NNS and_CC in_IN patients_NNS with_IN sepsis_NN or_CC septic_JJ shock_NN ._. 
SUBJECTS_NNS :_: Fifteen_CD patients_NNS (_( age_NN 25-79_CD )_) with_IN sepsis_NN or_CC septic_JJ shock_NN who_WP were_VBD admitted_VBN to_TO an_DT intensive_JJ care_NN unit_NN were_VBD studied_VBN ._. 
The_DT control_JJ group_NN consisted_VBD of_IN 24_CD healthy_JJ laboratory_NN employees_NNS ._. 
MEASUREMENTS_NNS :_: The_DT binding_NN capacity_NN and_CC affinity_NN of_IN the_DT glucocorticoid_NN receptors_NNS were_VBD measured_VBN and_CC compared_VBN to_TO clinical_JJ data_NNS and_CC the_DT plasma_NN cortisol_NN concentrations_NNS ._. 
RESULTS_NNS :_: Hypercortisolaemia_NN ,_, in_FW vitro_FW ,_, resulted_VBD in_IN a_DT decreased_VBN affinity_NN and_CC a_DT decreased_VBN binding_NN capacity_NN of_IN the_DT glucocorticoid_NN receptor_NN ._. 
In_FW vitro_FW ,_, hyperthermia_NN as_RB well_RB as_IN variations_NNS in_IN the_DT cellular_JJ composition_NN did_VBD not_RB influence_VB the_DT glucocorticoid_NN receptor_NN ._. 
In_FW vivo_FW ,_, there_EX was_VBD no_DT change_NN in_IN the_DT number_NN of_IN receptors_NNS per_IN cell_NN in_IN patients_NNS with_IN sepsis_NN or_CC septic_JJ shock_NN as_IN compared_VBN to_TO healthy_JJ controls_NNS ._. 
However_RB ,_, a_DT decreased_VBN affinity_NN of_IN the_DT glucocorticoid_NN receptor_NN was_VBD observed_VBN ._. 
There_EX was_VBD a_DT weak_JJ but_CC significant_JJ negative_JJ correlation_NN between_IN body_NN temperature_NN and_CC the_DT number_NN of_IN glucocorticoid_NN receptors_NNS in_IN the_DT patient_NN group_NN ._. 
There_EX was_VBD no_DT relation_NN between_IN circulating_VBG cortisol_NN concentrations_NNS and_CC glucocorticoid_NN receptor_NN affinity_NN and_CC number_NN ._. 
CONCLUSIONS_NNS :_: There_EX is_VBZ no_DT obvious_JJ regulation_NN of_IN the_DT number_NN of_IN glucocorticoid_NN receptors_NNS by_IN plasma_NN cortisol_NN concentrations_NNS in_FW vivo_FW ._. 
The_DT decreased_VBN affinity_NN of_IN the_DT glucocorticoid_NN receptor_NN together_RB with_IN the_DT negative_JJ correlation_NN between_IN hyperthermia_NN and_CC the_DT number_NN of_IN glucocorticoid_NN receptors_NNS in_IN patients_NNS with_IN sepsis_NN or_CC septic_JJ shock_NN suggest_VBP that_IN hypothalamic-pituitary-adrenal_JJ axis_NN activation_NN during_IN critical_JJ illness_NN is_VBZ accompanied_VBN by_IN peripheral_JJ adaptation_NN in_IN glucocorticoid_NN receptor_NN number_NN and_CC affinity_NN ._. 
Integrin-mediated_JJ tyrosine_NN phosphorylation_NN and_CC cytokine_NN message_NN induction_NN in_IN monocytic_JJ cells_NNS ._. 
A_DT possible_JJ signaling_NN role_NN for_IN the_DT Syk_NN tyrosine_NN kinase_NN ._. 
Activation_NN of_IN cytoplasmic_JJ tyrosine_NN kinases_NNS is_VBZ an_DT important_JJ aspect_NN of_IN signal_NN transduction_NN mediated_VBN by_IN integrins_NNS ._. 
In_IN the_DT human_JJ monocytic_JJ cell_NN line_NN THP-1_NN ,_, either_CC integrin_* -dependent_JJ cell_NN adhesion_NN to_TO fibronectin_NN or_CC ligation_NN of_IN beta_NN 1_CD integrins_NNS with_IN antibodies_NNS causes_VBZ a_DT rapid_JJ and_CC intense_JJ tyrosine_NN phosphorylation_NN of_IN two_CD sets_NNS of_IN proteins_NNS of_IN about_RB 65-75_CD and_CC 120-125_CD kDa_NN ._. 
In_IN addition_NN ,_, integrin_NN ligation_NN leads_VBZ to_TO nuclear_JJ translocation_NN of_IN the_DT p50_NN and_CC p65_NN subunits_NNS of_IN the_DT NF-kappa_NN B_NN transcription_NN factor_NN ,_, to_TO activation_NN of_IN a_DT reporter_NN gene_NN driven_VBN by_IN a_DT promoter_NN containing_VBG NF-kappa_NN B_NN sites_NNS ,_, and_CC to_TO increased_VBN levels_NNS of_IN mRNAs_NNS for_IN immediate-early_JJ genes_NNS ,_, including_VBG the_DT cytokine_NN interleukin_NN (_( IL_NN )_) -1_CD beta_NN ._. 
The_DT tyrosine_NN kinase_NN inhibitors_NNS genistein_NN and_CC herbimycin_NN A_NN block_VBP both_CC integrin_* -mediated_JJ tyrosine_NN phosphorylation_NN and_CC increases_NNS in_IN IL-1_NN beta_NN message_NN levels_NNS ,_, indicating_VBG a_DT causal_JJ relationship_NN between_IN the_DT two_CD events_NNS ._. 
The_DT components_NNS tyrosine_NN phosphorylated_VBN subsequent_JJ to_TO cell_NN adhesion_NN include_VBP paxillin_NN ,_, pp125FAK_NN ,_, and_CC the_DT SH2_NN domain_NN containing_VBG tyrosine_NN kinase_NN Syk_NN ._. 
In_IN contrast_NN ,_, integrin_NN ligation_NN with_IN antibodies_NNS induces_VBZ tyrosine_NN phosphorylation_NN of_IN Syk_NN but_CC not_RB of_IN FAK_NN or_CC paxillin_NN ._. 
In_IN adhering_JJ cells_NNS ,_, pre-treatment_NN with_IN cytochalasin_NN D_NN suppresses_VBZ tyrosine_NN phosphorylation_NN of_IN FAK_NN and_CC paxillin_NN but_CC not_RB of_IN Syk_NN ,_, while_IN IL-1_NN beta_NN message_NN induction_NN is_VBZ unaffected_JJ ._. 
These_DT observations_NNS indicate_VBP that_IN the_DT Syk_NN tyrosine_NN kinase_NN may_MD be_VB an_DT important_JJ component_NN of_IN an_DT integrin_NN signaling_NN pathway_NN in_IN monocytic_JJ cells_NNS ,_, leading_VBG to_TO activation_NN of_IN NF-kappa_NN B_NN and_CC to_TO increased_VBN levels_NNS of_IN cytokine_NN messages_NNS ._. 
Inhibition_NN of_IN dexamethasone_NN binding_NN to_TO human_JJ glucocorticoid_NN receptor_NN by_IN New_NN World_NN primate_JJ cell_NN extracts_NNS ._. 
To_TO determine_VB if_IN New_NN World_NN primates_NNS express_VBP an_DT inhibitor_NN that_WDT influences_VBZ glucocorticoid_NN receptor_NN (_( GR_NN )_) binding_NN characteristics_NNS ,_, we_PRP examined_VBD [3H]dexamethasone_NN binding_NN in_IN cytosol_NN prepared_VBN from_IN B95-8_NN lymphoid_JJ cells_NNS ,_, derived_VBN from_IN the_DT cotton_NN top_NN tamarin_NN (_( Saguinus_FW oedipus_FW )_) ,_, in_IN combination_NN with_IN cytosol_NN prepared_VBN from_IN human_JJ or_CC rat_NN tissues_NNS ._. 
B95-8_NN cytosol_NN inhibited_VBD specific_JJ binding_NN of_IN [3H]dexamethasone_NN (_( P_NN <_JJR 0.01_CD )_) when_WRB mixed_VBN with_IN cytosol_NN prepared_VBN from_IN either_CC a_DT human_JJ lymphoid_JJ cell_NN line_NN (_( HL_NN )_) or_CC rat_NN thymus_NN ._. 
The_DT inhibitory_JJ activity_NN was_VBD heat_NN labile_JJ and_CC trypsin_NN sensitive_JJ ._. 
Peak_JJ inhibitory_JJ activity_NN was_VBD found_VBN in_IN the_DT 150-200_CD kd_NN fractions_NNS after_IN Sephacryl_NN G-200_NN ultrafiltration_NN ._. 
Scatchard_NN analysis_NN of_IN [3H]dexamethasone_NN binding_NN using_VBG mixed_JJ cytosol_NN showed_VBD a_DT diminished_VBN GR_NN apparent_JJ binding_NN affinity_NN when_WRB compared_VBN to_TO HL_NN cytosol_NN ._. 
Kinetic_JJ studies_NNS using_VBG mixed_JJ cytosol_NN indicated_VBD that_IN B95-8_NN cytosol_NN did_VBD not_RB affect_VB the_DT apparent_JJ dissociation_NN rate_NN of_IN [3H]dexamethasone_NN ._. 
These_DT data_NNS demonstrate_VBP that_IN B95-8_NN cells_NNS contain_VBP a_DT competitive_JJ inhibitor_NN that_WDT prevents_VBZ binding_NN of_IN dexamethasone_NN to_TO its_PRP$ cognate_JJ receptor_NN ._. 
Disruption_NN of_IN a_DT GATA_NN motif_NN in_IN the_DT Duffy_NN gene_NN promoter_NN abolishes_VBZ erythroid_JJ gene_NN expression_NN in_IN Duffy-negative_JJ individuals_NNS ._. 
The_DT mRNA_NN for_IN the_DT Duffy_NN blood_NN group_NN antigen_NN ,_, the_DT erythrocyte_NN receptor_NN for_IN the_DT Plasmodium_NN vivax_NN malaria_NN parasite_NN ,_, has_VBZ recently_RB been_VBN cloned_VBN and_CC shown_VBN to_TO encode_VB a_DT widely_RB expressed_VBN chemokine_NN receptor_NN ._. 
Here_RB ,_, we_PRP show_VBP that_IN the_DT Duffy_NN antigen/chemokine_NN receptor_NN gene_NN (_( DARC_NN )_) is_VBZ composed_VBN of_IN a_DT single_JJ exon_NN and_CC that_IN most_JJS Duffy-negative_JJ blacks_NNS carry_VBP a_DT silent_JJ FY*B_NN allele_NN with_IN a_DT single_JJ T_NN to_TO C_NN substitution_NN at_IN nucleotide_NN -46_CD ._. 
This_DT mutation_NN impairs_VBZ the_DT promoter_NN activity_NN in_IN erythroid_JJ cells_NNS by_IN disrupting_VBG a_DT binding_VBG site_NN for_IN the_DT GATA1_NN erythroid_JJ transcription_NN factor_NN ._. 
With_IN the_DT recent_JJ characterization_NN of_IN the_DT FY*A_NN and_CC FY*B_NN alleles_NNS ,_, these_DT findings_NNS provide_VBP the_DT molecular_JJ basis_NN of_IN the_DT Duffy_NN blood_NN group_NN system_NN and_CC an_DT explanation_NN for_IN the_DT erythroid-specific_JJ repression_NN of_IN the_DT DARC_NN gene_NN in_IN Duffy-negative_JJ individuals_NNS ._. 
Activation_NN of_IN pp90rsk_NN and_CC early_JJ growth_NN response-1_NN gene_NN expression_NN by_IN pokeweed_NN mitogen_NN in_IN human_JJ B_NN cells_NNS ._. 
The_DT present_JJ studies_NNS have_VBP examined_VBN the_DT effects_NNS of_IN pokeweed_NN mitogen_NN (_( PWM_NN )_) on_IN the_DT induction_NN of_IN early_JJ growth_NN response-1_NN gene_NN (_( EGR-1_NN )_) in_IN normal_JJ human_JJ B_NN cells_NNS ._. 
PWM_NN regulates_VBZ EGR-1_NN gene_NN expression_NN by_IN both_CC transcriptional_JJ and_CC post-transcriptional_JJ mechanisms_NNS ._. 
Transient_JJ transfection_NN assays_NNS with_IN EGR-1_NN promoter_NN fragments_NNS linked_VBN to_TO the_DT chloramphenicol_NN acetyltransferase_NN (_( CAT_NN )_) gene_NN demonstrated_VBD that_IN PWM_NN induced_JJ EGR-1_NN transcription_NN is_VBZ conferred_VBN by_IN the_DT CArG_NN motif_NN (_( C_NN C[AT]6GG_NN )_) in_IN the_DT EGR-1_NN promoter_NN ._. 
The_DT results_NNS further_RB demonstrated_VBD the_DT activation_NN of_IN S6_NN kinase_NN (_( pp90rsk_NN )_) ,_, evidenced_VBN by_IN phosphorylation_NN of_IN S6_NN and_CC serum_NN response_NN factor_NN (_( SRF_NN )_) peptides_NNS ,_, in_IN PWM_NN treated_JJ B_NN cells_NNS ._. 
Taken_VBN together_RB ,_, these_DT findings_NNS suggest_VBP that_IN PWM_NN is_VBZ able_JJ to_TO initiate_VB an_DT intracytoplasmic_JJ signalling_NN cascade_NN and_CC EGR-1_NN induction_NN in_IN normal_JJ human_JJ B_NN cells_NNS ._. 
A_DT conserved_VBN motif_NN in_IN the_DT promoters_NNS of_IN several_JJ cytokines_NNS expressed_VBN by_IN human_JJ Th2-type_JJ lymphocytes_NNS ._. 
We_PRP have_VBP recently_RB found_VBN a_DT novel_JJ conserved_VBN motif_NN in_IN the_DT promoters_NNS of_IN several_JJ T-cell-expressed_JJ cytokines_NNS [_( human_JJ interleukin_* -2_NN ,_, -4_CD ,_, -5_CD and_CC -13_CD and_CC human_JJ and_CC mouse_NN granulocyte/macrophage-colony_NN stimulating_JJ factor_NN (_( GM-CSF_NN )_) ]_) ._. 
It_PRP contains_VBZ a_DT core_NN sequence_NN CTTGG_NN ..._: CCAAG_NN which_WDT is_VBZ present_JJ as_IN part_NN of_IN larger_JJR palindromic_JJ sequences_NNS in_IN each_DT gene_NN ._. 
This_DT suggest_VBP that_IN they_PRP may_MD interact_VB with_IN a_DT new_JJ family_NN of_IN trans-acting_JJ factors_NNS ._. 
In_IN transfection_NN assays_NNS ,_, the_DT human_JJ GM-CSF_NN element_NN has_VBZ a_DT strong_JJ positive_JJ effect_NN on_IN the_DT expression_NN of_IN a_DT reporter_NN gene_NN by_IN the_DT human_JJ T_NN cell_NN line_NN Jurkat_NN J6_NN upon_IN stimulation_NN ._. 
In_IN DNA_NN mobility_NN shift_NN assays_NNS ,_, this_DT sequence_NN can_MD give_VB either_CC six_CD different_JJ specific_JJ bands_NNS which_WDT are_VBP competed_VBN out_RP by_IN different_JJ parts_NNS of_IN the_DT sequence_NN or_CC one_CD specific_JJ band_NN which_WDT is_VBZ competed_VBN out_RP by_IN each_DT of_IN the_DT inverted_VBN repeats_NNS ,_, depending_VBG on_IN the_DT reconstitution_NN conditions_NNS ._. 
In_IN different_JJ genes_NNS ,_, the_DT core_NN sequences_NNS are_VBP separated_VBN by_IN integer_NN numbers_NNS of_IN helical_JJ turns_NNS ._. 
Considering_VBG the_DT strong_JJ positive_JJ regulatory_JJ effect_NN of_IN this_DT element_NN and_CC its_PRP$ presence_NN in_IN several_JJ T-cell-expressed_JJ cytokine_NN genes_NNS ,_, it_PRP may_MD be_VB crucial_JJ to_TO the_DT coordinated_VBN expression_NN of_IN these_DT cytokines_NNS in_IN T_NN helper_NN cells_NNS ._. 
cDNA_NN cloning_NN of_IN a_DT NGFI-B/nur77-related_JJ transcription_NN factor_NN from_IN an_DT apoptotic_JJ human_JJ T_NN cell_NN line_NN ._. 
A_DT human_JJ T_NN lymphoid_JJ cell_NN line_NN ,_, PEER_NN ,_, dies_VBZ by_IN apoptosis_NN in_IN the_DT presence_NN of_IN PMA_NN and_CC calcium_NN ionophore_NN ._. 
A_DT new_JJ gene_NN ,_, TINUR_NN ,_, was_VBD cloned_VBN from_IN apoptotic_JJ PEER_NN cells_NNS ._. 
The_DT expression_NN of_IN the_DT TINUR_NN gene_NN is_VBZ induced_VBN within_IN 1_CD h_NN after_IN the_DT cross-linking_NN of_IN the_DT T_NN cell_NN Ag_NN receptor_NN complex_NN ._. 
TINUR_NN belongs_VBZ to_TO the_DT NGFI-B/nur77_NN family_NN of_IN the_DT steroid_NN receptor_NN superfamily_NN and_CC is_VBZ an_DT orphan_NN receptor_NN ._. 
TINUR_NN binds_VBZ to_TO the_DT same_JJ DNA_NN sequence_NN as_IN NGFI-B/nur77_NN ._. 
We_PRP also_RB propose_VBP that_IN the_DT NGFI-B/nur77_NN family_NN can_MD be_VB classified_VBN into_IN two_CD subtypes_NNS ._. 
Aspirin_NN inhibits_VBZ nuclear_JJ factor-kappa_NN B_NN mobilization_NN and_CC monocyte_NN adhesion_NN in_IN stimulated_VBN human_JJ endothelial_JJ cells_NNS ._. 
BACKGROUND_NN :_: The_DT induction_NN of_IN vascular_JJ cell_NN adhesion_NN molecule-1_NN (_( VCAM-1_NN )_) and_CC E-selectin_NN by_IN tumor_NN necrosis_NN factor-alpha_NN (_( TNF_NN )_) is_VBZ mediated_VBN by_IN mobilization_NN of_IN the_DT transcription_NN factor_NN nuclear_JJ factor-kappa_NN B_NN (_( NF-kappa_NN B_NN )_) ._. 
Since_IN salicylates_NNS have_VBP been_VBN reported_VBN to_TO inhibit_VB NF-kappa_NN B_NN activation_NN by_IN preventing_VBG the_DT degradation_NN of_IN its_PRP$ inhibitor_NN I_NN kappa_NN B_NN ,_, we_PRP studied_VBD a_DT potential_JJ inhibition_NN of_IN this_DT pathway_NN by_IN acetylsalicylate_NN (_( aspirin_NN )_) in_IN human_JJ umbilical_JJ vein_NN endothelial_JJ cells_NNS (_( HUVECs_NNS )_) ._. 
METHODS_NNS AND_CC RESULTS_NNS :_: Gel-shift_JJ analyses_NNS demonstrated_VBD dose-dependent_JJ inhibition_NN of_IN TNF_* -induced_JJ NF-kappa_NN B_NN mobilization_NN by_IN aspirin_NN at_IN concentrations_NNS ranging_VBG from_IN 1_CD to_TO 10_CD mmol/L_NN ._. 
Induction_NN of_IN VCAM-1_NN and_CC E-selectin_NN surface_NN expression_NN by_IN TNF_NN was_VBD dose-dependently_RB reduced_VBN by_IN aspirin_NN over_IN the_DT same_JJ range_NN ,_, while_IN induction_NN of_IN intercellular_JJ adhesion_NN molecule-1_NN (_( ICAM-1_NN )_) was_VBD hardly_RB affected_VBN ._. 
Aspirin_NN appeared_VBD to_TO prevent_VB VCAM-1_NN transcription_NN ,_, since_IN it_PRP dose-dependently_RB inhibited_VBD induction_NN of_IN VCAM-1_NN mRNA_NN by_IN TNF_NN ._. 
As_IN a_DT functional_JJ consequence_NN ,_, adhesion_NN of_IN U937_NN monocytes_NNS to_TO TNF_* -stimulated_JJ HUVECs_NNS was_VBD markedly_RB reduced_VBN by_IN aspirin_NN due_JJ to_TO suppression_NN of_IN VCAM-1_NN and_CC E-selectin_NN upregulation_NN ._. 
These_DT effects_NNS of_IN aspirin_NN were_VBD not_RB related_JJ to_TO the_DT inhibition_NN of_IN cyclooxygenase_NN activity_NN ,_, since_IN indomethacin_NN was_VBD ineffective_JJ ._. 
CONCLUSIONS_NNS :_: Our_PRP$ data_NNS suggest_VBP that_IN aspirin_NN inhibits_VBZ NF-kappa_NN B_NN mobilization_NN ,_, induction_NN of_IN VCAM-1_NN and_CC E-selectin_NN ,_, and_CC subsequent_JJ monocyte_NN adhesion_NN in_IN endothelial_JJ cells_NNS stimulated_VBN by_IN TNF_NN ,_, thereby_RB providing_VBG an_DT additional_JJ mechanism_NN for_IN therapeutic_JJ effects_NNS of_IN aspirin_NN ._. 
Activation_NN of_IN NF-kappa_NN B_NN by_IN phosphatase_NN inhibitors_NNS involves_VBZ the_DT phosphorylation_NN of_IN I_NN kappa_NN B_NN alpha_NN at_IN phosphatase_NN 2A-sensitive_JJ sites_NNS ._. 
Activation_NN of_IN NF-kappa_NN B_NN by_IN various_JJ cellular_JJ stimuli_NNS involves_VBZ the_DT phosphorylation_NN and_CC subsequent_JJ degradation_NN of_IN its_PRP$ inhibitor_NN ,_, I_NN kappa_NN B_NN alpha_NN ,_, although_IN the_DT underlying_VBG mechanism_NN remains_VBZ unclear_JJ ._. 
In_IN the_DT present_JJ study_NN ,_, the_DT role_NN of_IN serine/threonine_NN phosphatases_NNS in_IN the_DT regulation_NN of_IN I_NN kappa_NN B_NN alpha_NN phosphorylation_NN was_VBD investigated_VBN ._. 
Our_PRP$ studies_NNS demonstrate_VBP that_IN incubation_NN of_IN human_JJ T_NN cells_NNS with_IN low_JJ concentrations_NNS (_( approximately_RB 1-5_CD nM_NN )_) of_IN calyculin_NN A_NN or_CC okadaic_JJ acid_NN ,_, potent_JJ inhibitors_NNS of_IN protein_NN phosphatase_NN type_NN 1_CD (_( PP-1_NN )_) and_CC type_NN 2A_NN (_( PP-2A_NN )_) ,_, induces_VBZ the_DT phosphorylation_NN of_IN I_NN kappa_NN B_NN alpha_NN even_RB in_IN the_DT absence_NN of_IN any_DT cellular_JJ stimulus_NN ._. 
This_DT action_NN of_IN the_DT phosphatase_NN inhibitors_NNS ,_, which_WDT is_VBZ associated_VBN with_IN the_DT activation_NN of_IN the_DT RelA.p50_NN NF-kappa_NN B_NN heterodimer_NN ,_, is_VBZ not_RB affected_VBN by_IN agents_NNS that_WDT block_VBP the_DT induction_NN of_IN I_NN kappa_NN B_NN alpha_NN phosphorylation_NN by_IN tumor_NN necrosis_NN factor_NN alpha_NN (_( TNF-alpha_NN )_) ._. 
Furthermore_RB ,_, the_DT phosphorylated_VBN I_NN kappa_NN B_NN alpha_NN from_IN calyculin_NN A_* -treated_JJ cells_NNS ,_, but_CC not_RB that_DT from_IN TNF-alpha_* -stimulated_JJ cells_NNS ,_, is_VBZ sensitive_JJ to_TO PP-2A_NN in_FW vitro_FW ,_, suggesting_VBG the_DT existence_NN of_IN fundamental_JJ differences_NNS in_IN the_DT phosphorylation_NN of_IN I_NN kappa_NN B_NN alpha_NN induced_VBN by_IN the_DT two_CD different_JJ NF-kappa_NN B_NN inducers_NNS ._. 
However_RB ,_, induction_NN of_IN I_NN kappa_NN B_NN alpha_NN phosphorylation_NN by_IN both_CC TNF-alpha_NN and_CC the_DT phosphatase_NN inhibitors_NNS is_VBZ associated_VBN with_IN the_DT subsequent_JJ degradation_NN of_IN I_NN kappa_NN B_NN alpha_NN ._. 
We_PRP further_RB demonstrate_VBP that_IN TNF-alpha-_NN and_CC calyculin_NN A-induced_JJ I_NN kappa_NN B_NN alpha_NN degradation_NN exhibits_VBZ similar_JJ but_CC not_RB identical_JJ sensitivities_NNS to_TO a_DT proteasome_JJ inhibitor_NN ._. 
Together_RB ,_, these_DT results_NNS suggest_VBP that_IN phosphorylation_NN of_IN I_NN kappa_NN B_NN alpha_NN ,_, mediated_VBN through_IN both_CC the_DT TNF-alpha_* -inducible_JJ and_CC the_DT PP-2A_* -opposing_JJ kinases_NNS ,_, may_MD serve_VB to_TO target_NN I_NN kappa_NN B_NN alpha_NN for_IN proteasome_* -mediated_JJ degradation_NN ._. 
Sp1_NN functions_VBZ in_IN a_DT chromatin_* -dependent_JJ manner_NN to_TO augment_VB human_JJ alpha-globin_NN promoter_NN activity_NN ._. 
The_DT 5'_JJ flanking_JJ region_NN of_IN the_DT human_JJ alpha-globin_NN gene_NN is_VBZ highly_RB rich_JJ and_CC contains_VBZ multiple_JJ copies_NNS of_IN the_DT consensus_NN sequence_NN for_IN the_DT Sp1_NN binding_NN site_NN ._. 
We_PRP investigated_VBD the_DT role_NN of_IN this_DT region_NN in_IN augmenting_VBG alpha-globin_NN promoter_NN activity_NN in_IN the_DT presence_NN of_IN the_DT far-upstream_JJ alpha-globin_NN enhancer_NN ,_, HS-40_NN ._. 
We_PRP show_VBP that_IN in_IN transiently_RB transfected_VBN erythroid_JJ cells_NNS ,_, deletion_NN of_IN the_DT alpha-globin_NN 5'_JJ flanking_JJ region_NN has_VBZ no_DT effect_NN on_IN alpha-globin_NN promoter_NN activity_NN ._. 
However_RB ,_, upon_IN stable_JJ integration_NN into_IN chromatin_NN ,_, deletion_NN of_IN this_DT region_NN causes_VBZ a_DT nearly_RB 90_CD %_NN decrease_NN in_IN promoter_NN activity_NN compared_VBN with_IN expression_NN from_IN an_DT alpha-globin_NN promoter_NN retaining_VBG this_DT region_NN ._. 
These_DT results_NNS suggest_VBP that_IN the_DT alpha-globin_NN 5'_JJ flanking_JJ region_NN augments_VBZ alpha-globin_NN promoter_NN activity_NN in_IN a_DT chromatin_* -dependent_JJ manner_NN ._. 
We_PRP further_RB show_VBP that_IN this_DT region_NN is_VBZ required_VBN for_IN the_DT activation_NN of_IN alpha-globin_NN gene_NN expression_NN during_IN erythroid_JJ differentiation_NN ._. 
Finally_RB ,_, we_PRP show_VBP by_IN both_CC footprint_NN analysis_NN and_CC functional_JJ assays_NNS that_IN the_DT ability_NN of_IN the_DT region_NN to_TO increase_VB alpha-globin_NN promoter_NN activity_NN from_IN a_DT stably_RB integrated_VBN alpha-globin_NN gene_NN is_VBZ mediated_VBN by_IN its_PRP$ multiple_JJ binding_VBG sites_NNS for_IN the_DT transcription_NN factor_NN Sp1_NN ._. 
Costimulation_NN of_IN human_JJ CD4+_JJ T_NN cells_NNS with_IN LFA-3_NN and_CC B7_NN induce_VBP distinct_JJ effects_NNS on_IN AP-1_NN and_CC NF-kappa_NN B_NN transcription_NN factors_NNS ._. 
We_PRP have_VBP earlier_RBR shown_VBN that_IN stimulation_NN of_IN human_JJ CD4+_JJ T_NN cells_NNS with_IN SEA_NN presented_VBN on_IN Chinese_JJ hamster_NN ovary_NN (_( CHO_NN )_) -DR_NN transfectants_NNS coexpressing_VBG either_CC B7_NN or_CC LFA-3_NN resulted_VBD in_IN distinct_JJ cytokine_NN profiles_NNS ._. 
We_PRP now_RB demonstrate_VBP that_IN B7_NN ,_, but_CC not_RB LFA-3_NN ,_, strongly_RB costimulated_VBD IL-2_NN transcription_NN and_CC mRNA_NN expression_NN in_IN CD4+_JJ T_NN cells_NNS ._. 
Maximal_JJ increase_NN in_IN IL-2_NN transcription_NN was_VBD recorded_VBN with_IN CHO-DR/_* B7_* /_* LFA-3_NN ,_, suggesting_VBG a_DT cooperative_JJ effect_NN of_IN B7_NN and_CC LFA-3_NN at_IN the_DT transcriptional_JJ level_NN ._. 
Gel-shift_JJ analysis_NN demonstrated_VBD that_IN stimulation_NN of_IN CD4+_JJ T_NN cells_NNS with_IN CHO-DR_NN and_CC staphylococcal_JJ enterotoxin_NN A_NN was_VBD sufficient_JJ to_TO induce_VB significant_JJ amounts_NNS of_IN NF-kappa_NN B_NN binding_NN proteins_NNS ,_, whereas_IN induction_NN of_IN AP-1_NN binding_NN proteins_NNS required_VBD costimulation_NN ._. 
LFA-3_NN induced_VBD moderate_JJ levels_NNS of_IN AP-1_NN ,_, but_CC did_VBD not_RB influence_VB the_DT levels_NNS of_IN NF-kappa_NN B_NN ,_, while_IN B7_NN costimulation_NN strongly_RB induced_VBD both_CC AP-1_NN and_CC substantially_RB enhanced_VBN NF-kappa_NN B_NN binding_NN proteins_NNS ._. 
The_DT CHO-DR/_* B7_* /_* LFA-3_* triple_JJ transfectant_NN induced_VBD a_DT further_JJ increase_NN in_IN AP-1_NN and_CC NF-kappa_NN B_NN binding_NN proteins_NNS compared_VBN with_IN the_DT double_JJ transfectants_NNS ._. 
The_DT level_NN of_IN Oct-1_NN binding_NN proteins_NNS remained_VBD similar_JJ in_IN all_DT samples_NNS ._. 
Super-shift_JJ analysis_NN revealed_VBD that_IN the_DT NF-kappa_NN B_NN complex_NN of_IN costimulated_VBN CD4+_JJ T_NN cells_NNS contained_VBD large_JJ amounts_NNS of_IN p50_NN ,_, substantial_JJ amounts_NNS of_IN p65_NN ,_, and_CC marginal_JJ levels_NNS of_IN c-Rel_NN proteins_NNS ._. 
The_DT AP-1_NN binding_NN proteins_NNS contained_VBD c-Jun_NN ,_, Jun-D_NN ,_, and_CC Fra-1_NN ,_, but_CC marginal_JJ amounts_NNS of_IN Jun-B_NN and_CC c-Fos_NN ._. 
Our_PRP$ results_NNS indicate_VBP distinct_JJ effects_NNS of_IN B7_NN and_CC LFA-3_NN costimulation_NN on_IN the_DT activity_NN of_IN AP-1_NN and_CC NF-kappa_NN B_NN ._. 
These_DT may_MD partly_RB account_VB for_IN the_DT differential_JJ effects_NNS of_IN B7_NN and_CC LFA-3_NN costimulation_NN on_IN IL-2_NN expression_NN ._. 
The_DT Ah_NN receptor_NN recognizes_VBZ DNA_NN binding_NN sites_NNS for_IN the_DT B_NN cell_NN transcription_NN factor_NN ,_, BSAP_NN :_: a_DT possible_JJ mechanism_NN for_IN dioxin-mediated_JJ alteration_NN of_IN CD19_NN gene_NN expression_NN in_IN human_JJ B_NN lymphocytes_NNS ._. 
2,3,7,8-tetrachlorodibenzo-p-dioxin_NN (_( TCDD_NN )_) inhibits_VBZ murine_JJ and_CC human_JJ B_NN lymphocyte_NN immunoglobulin_NN production_NN through_IN an_DT unknown_JJ mechanism_NN ._. 
This_DT study_NN investigated_VBD the_DT effect_NN of_IN TCDD_NN on_IN expression_NN of_IN the_DT CD19_NN gene_NN in_IN a_DT human_JJ B_NN lymphocyte_NN cell_NN line_NN ._. 
Northern_NN blot_NN analysis_NN showed_VBD that_IN TCDD_NN treatment_NN decreased_VBD steady_JJ state_NN levels_NNS of_IN CD19_NN mRNA_NN by_IN 67_CD %_NN in_IN the_DT IM-9_NN cell_NN line_NN ._. 
Using_VBG a_DT gel_NN mobility_NN shift_NN assay_NN ,_, we_PRP identified_VBD a_DT DNA-binding_NN complex_NN in_IN IM-9_NN nuclear_JJ extracts_NNS that_WDT by_IN several_JJ criteria_NNS appears_VBZ to_TO be_VB the_DT Ah_NN receptor_NN ._. 
In_IN addition_NN ,_, the_DT Ah_NN receptor_NN complex_NN recognized_VBD a_DT DNA_NN binding_NN site_NN for_IN B_NN cell_NN lineage-specific_JJ activator_NN protein_NN (_( BSAP_NN )_) in_IN the_DT promoter_NN region_NN of_IN the_DT human_JJ CD19_NN gene_NN which_WDT is_VBZ similar_JJ to_TO the_DT consensus_NN Ah_NN receptor_NN DNA_NN binding_NN site_NN ._. 
These_DT results_NNS suggest_VBP that_IN the_DT AhR_NN could_MD interfere_VB with_IN BSAP_* -stimulated_JJ CD19_NN gene_NN transcription_NN by_IN competition_NN for_IN a_DT common_JJ DNA_NN binding_NN site_NN ._. 
Inhibitory_JJ action_NN of_IN nm23_NN proteins_NNS on_IN induction_NN of_IN erythroid_JJ differentiation_NN of_IN human_JJ leukemia_NN cells_NNS ._. 
We_PRP recently_RB identified_VBD a_DT differentiation_NN inhibitory_JJ factor_NN (_( I-factor_NN )_) in_IN mouse_NN myeloid_JJ leukemia_NN M1_NN cells_NNS as_IN a_DT murine_JJ homolog_NN of_IN the_DT human_JJ nm23-H2_NN gene_NN product_NN ._. 
nm23_NN genes_NNS encode_VBP proteins_NNS that_WDT participate_VBP in_IN tumor_NN metastasis_NN regulation_NN and_CC in_IN various_JJ fundamental_JJ cellular_JJ processes_NNS ,_, although_IN their_PRP$ mechanisms_NNS of_IN action_NN are_VBP still_RB unknown_JJ ._. 
Although_IN all_DT nm23_NN proteins_NNS contain_VBP nucleoside_NN diphosphate_NN (_( NDP_NN )_) kinase_NN activity_NN ,_, it_PRP has_VBZ not_RB been_VBN established_VBN that_IN the_DT enzyme_NN activity_NN mediated_VBD the_DT various_JJ functions_NNS of_IN nm23_NN proteins_NNS ._. 
In_IN the_DT present_JJ experiment_NN ,_, we_PRP examined_VBD the_DT effect_NN of_IN nm23_NN proteins_NNS on_IN various_JJ differentiation_NN induction_NN systems_NNS of_IN human_JJ leukemic_JJ cells_NNS including_VBG HL-60_NN ,_, U937_NN ,_, HEL/S_NN ,_, KU812F_NN ,_, K562_NN ,_, and_CC HEL_NN cells_NNS ._. 
Native_JJ human_JJ erythrocyte_NN NDP_NN kinase_NN protein_NN inhibited_VBD the_DT induction_NN of_IN erythroid_JJ differentiation_NN of_IN HEL_NN ,_, KU812_NN and_CC K562_NN cells_NNS ,_, but_CC not_RB the_DT induction_NN of_IN monocytic_JJ or_CC granulocytic_JJ differentiation_NN of_IN HL-60_NN ,_, U937_NN and_CC HEL/S_NN cells_NNS ._. 
The_DT erythroid_JJ differentiation_NN of_IN HEL_NN cells_NNS was_VBD inhibited_VBN by_IN recombinant_JJ human_JJ nm23_* -H1_NN ,_, -H2_NN ,_, mouse_NN nm23_* -M1_NN ,_, and_CC -M2_NN proteins_NNS ._. 
Moreover_RB ,_, both_CC the_DT mutant_JJ nm23-H2His_NN protein_NN and_CC truncated_VBN nm23-H2_NN protein_NN containing_VBG N-terminal_JJ (_( 1-60_CD )_) peptide_NN ,_, which_WDT do_VBP not_RB have_VB NDP_NN kinase_NN activity_NN ,_, also_RB inhibited_VBD erythroid_JJ differentiation_NN of_IN HEL_NN cells_NNS ._. 
These_DT results_NNS suggest_VBP that_IN (_( 1_LS )_) the_DT differentiation_NN inhibitory_JJ activity_NN of_IN I-factor_* /nm23_NN protein_NN is_VBZ not_RB restricted_JJ to_TO monocytic_JJ differentiation_NN of_IN M1_NN cells_NNS ,_, (_( 2_LS )_) the_DT inhibitory_JJ activity_NN is_VBZ exhibited_VBN without_IN species_NNS specificity_NN ,_, and_CC (_( 3_LS )_) the_DT differentiation_NN inhibitory_JJ activity_NN of_IN the_DT nm23/NDP_NN kinase_NN protein_NN is_VBZ independent_JJ of_IN its_PRP$ enzyme_NN activity_NN and_CC requires_VBZ the_DT presence_NN of_IN N-terminal_JJ peptides_NNS ._. 
Lipopolysaccharide_* -induced_JJ E-selectin_NN expression_NN requires_VBZ continuous_JJ presence_NN of_IN LPS_NN and_CC is_VBZ inhibited_VBN by_IN bactericidal/permeability-increasing_JJ protein_NN ._. 
Endothelial_JJ cells_NNS stimulated_VBN by_IN LPS_NN express_VBP E-selectin_NN ,_, which_WDT plays_VBZ an_DT important_JJ role_NN in_IN mediating_VBG neutrophil_NN adhesion_NN during_IN inflammation_NN ._. 
E-selectin_NN is_VBZ induced_VBN within_IN 1-2_CD h_NN ,_, peaks_VBZ at_IN 4-6_CD h_NN ,_, and_CC gradually_RB returns_VBZ to_TO basal_JJ level_NN by_IN 24_CD h_NN ._. 
rBPI21_NN ,_, a_DT recombinant_JJ N-terminal_JJ fragment_NN of_IN human_JJ bactericidal/permeability-increasing_JJ protein_NN (_( BPI_NN )_) ,_, inhibited_VBD LPS_* -induced_JJ E-selectin_NN expression_NN when_WRB added_VBN at_IN the_DT same_JJ time_NN as_IN ,_, and_CC up_IN to_TO 6_CD h_NN after_IN ,_, LPS_NN ._. 
Delayed_VBN administration_NN of_IN rBPI21_NN also_RB affected_VBD LPS_* -mediated_JJ activation_NN of_IN the_DT nuclear_JJ factor_NN ,_, NF-kappa_NN B_NN ._. 
Two_CD to_TO 4_CD h_NN following_VBG LPS_NN addition_NN to_TO endothelial_JJ cells_NNS ,_, when_WRB NF-kappa_NN B_NN was_VBD already_RB activated_VBN ,_, addition_NN of_IN rBPI21_NN resulted_VBD in_IN marked_JJ reduction_NN of_IN NF-kappa_NN B_NN detectable_JJ at_IN 4_CD or_CC 6_CD h_NN ._. 
These_DT results_NNS indicate_VBP that_IN endothelial_JJ activation_NN requires_VBZ continuous_JJ presence_NN of_IN LPS_NN ,_, and_CC rBPI21_NN acts_VBZ to_TO reverse_VB LPS_* -mediated_JJ endothelial_JJ activation_NN by_IN interrupting_VBG the_DT on-going_JJ LPS_NN signal_NN ._. 
GM-CSF_NN and_CC IL-2_NN share_VBP common_JJ control_NN mechanisms_NNS in_IN response_NN to_TO costimulatory_JJ signals_NNS in_IN T_NN cells_NNS ._. 
Antigen_NN complexed_VBN with_IN major_JJ histocompatibility_NN complex_NN class_NN I_CD or_CC II_CD molecules_NNS on_IN the_DT surface_NN of_IN antigen_NN presenting_NN cells_NNS interacts_VBZ with_IN the_DT T_NN cell_NN receptor_NN (_( TCR_NN )_) on_IN the_DT surface_NN of_IN T_NN cells_NNS and_CC initiates_VBZ an_DT activation_NN cascade_NN ._. 
So_RB called_VBN costimulatory_JJ signals_NNS ,_, mediated_VBN by_IN other_JJ cell_NN surface_NN interactions_NNS or_CC soluble_JJ cytokines_NNS produced_VBN by_IN antigen_NN presenting_NN cells_NNS ,_, are_VBP also_RB required_VBN for_IN complete_JJ T_NN cell_NN activation_NN ._. 
High_JJ levels_NNS of_IN cytokine_NN gene_NN expression_NN in_IN T_NN cells_NNS also_RB required_VBD both_CC TCR_NN and_CC costimulatory_JJ signals_NNS ._. 
The_DT granulocyte-macrophage_JJ colony-stimulating_JJ factor_NN requires_VBZ sequences_NNS in_IN the_DT promoter_NN as_RB well_RB as_IN a_DT powerful_JJ enhancer_NN located_JJ 3kb_RB upstream_RB to_TO respond_VB to_TO TCR_* -like_JJ signals_NNS ._. 
These_DT promoter_NN and_CC enhancer_NN regions_NNS are_VBP mainly_RB activated_VBN by_IN the_DT transcription_NN factor_NN nuclear_JJ factor_NN of_IN activated_VBN T_NN cells_NNS (_( NFAT_NN )_) ._. 
The_DT activation_NN of_IN NFAT_NN by_IN TCR_NN signals_NNS has_VBZ been_VBN well_RB described_VBN for_IN interleukin-2_NN (_( IL-2_NN )_) and_CC IL-4_NN gene_NN transcription_NN in_IN T_NN cells_NNS ._. 
Costimulatory_JJ signals_NNS ,_, such_JJ as_IN activation_NN of_IN the_DT CD28_NN cell_NN surface_NN molecule_NN on_IN T_NN cells_NNS ,_, lead_VBP to_TO activation_NN through_IN a_DT distinct_JJ region_NN of_IN the_DT granulocyte-macrophage_JJ colony-stimulating_JJ factor_NN (_( GM-CSF_NN )_) promoter_NN ._. 
This_DT region_NN is_VBZ termed_VBN the_DT CK-1_NN or_CC CD28RE_NN and_CC appears_VBZ to_TO bind_VB specific_JJ members_NNS of_IN the_DT NF-kappa_NN B_NN family_NN of_IN transcription_NN factors_NNS ._. 
Human_JJ T_NN leukemia_NN virus_NN type_NN 1_CD (_( HTLV-1_NN )_) infects_VBZ T_NN cells_NNS and_CC can_MD lead_VB to_TO increase_VB GM-CSF_NN expression_NN ._. 
We_PRP have_VBP found_VBN that_IN the_DT HTLV-1_NN transactivator_NN protein_NN ,_, tax_NN ,_, acts_VBZ as_IN a_DT costimulatory_JJ signal_NN for_IN GM-CSF_NN and_CC IL-2_NN gene_NN transcription_NN ,_, in_IN that_IN it_PRP can_MD cooperate_VB with_IN TCR_NN signals_NNS to_TO mediate_VB high_JJ level_NN gene_NN expression_NN ._. 
Tax_NN activates_VBZ the_DT GM-CSF_NN promoter_NN through_IN the_DT CK-1_* /CD28RE_NN region_NN and_CC also_RB activates_VBZ nuclear_JJ factor-kappa_NN B_NN binding_VBG to_TO this_DT region_NN ._. 
However_RB ,_, other_JJ transcription_NN factors_NNS or_CC coactivators_NNS of_IN NF-kappa_NN B_NN are_VBP required_VBN for_IN tax_NN activation_NN but_CC these_DT remain_VBP to_TO be_VB identified_VBN ._. 
The_DT CK-1_* /CD28RE_NN of_IN GM-CSF_NN shows_VBZ a_DT high_JJ degree_NN of_IN similarity_NN to_TO the_DT IL-2_NN CD28RE_NN and_CC the_DT IL-3_NN gene_NN also_RB contains_VBZ a_DT related_JJ region_NN ._. 
This_DT observation_NN ,_, together_RB with_IN the_DT fact_NN that_IN both_CC GM-CSF_NN and_CC IL-2_NN respond_VBP to_TO TCR_NN signals_NNS via_IN NFAT_NN ,_, implies_VBZ a_DT high_JJ degree_NN of_IN conservation_NN in_IN the_DT regulation_NN of_IN cytokine_NN gene_NN expression_NN in_IN T_NN cells_NNS ._. 
Danazol_NN decreases_VBZ transcription_NN of_IN estrogen_NN receptor_NN gene_NN in_IN human_JJ monocytes_NNS ._. 
1._LS Administration_NN of_IN danazol_NN for_IN over_IN one_CD month_NN reduced_VBD the_DT levels_NNS of_IN estrogen_NN receptor_NN (_( ER_NN )_) and_CC its_PRP$ mRNA_NN to_TO approximately_RB 50_CD and_CC 20_CD %_NN ,_, respectively_RB in_IN monocytes_NNS ._. 
2._LS Danazol_NN did_VBD not_RB alter_VB the_DT degradation_NN rate_NN of_IN ER_NN mRNA_NN in_IN monocytes_NNS ._. 
3._LS Danazol_NN decreased_VBD the_DT transcription_NN rate_NN of_IN ER_NN gene_NN to_TO approximately_RB 50_CD %_NN in_IN monocytes_NNS in_IN a_DT run-on_JJ assay_NN ._. 
4._LS Danazol_NN may_MD release_VB estrogen_NN predominance_NN via_IN the_DT reduction_NN of_IN transcription_NN for_IN ER_NN gene_NN ,_, which_WDT leads_VBZ to_TO the_DT reduction_NN of_IN ER_NN mRNA_NN and_CC ER_NN expressions_NNS in_IN monocytes_NNS ._. 
Functional_JJ characterization_NN of_IN novel_JJ IL-2_NN transcriptional_JJ inhibitors_NNS ._. 
IL-2_* -mediated_JJ T_NN cell_NN proliferation_NN is_VBZ a_DT critical_JJ early_JJ event_NN in_IN the_DT inflammatory_JJ process_NN ._. 
Formation_NN of_IN the_DT NFAT_* -1_NN transcriptional_JJ complex_NN on_IN the_DT IL-2_NN promoter_NN is_VBZ essential_JJ for_IN IL-2_NN transcription_NN ._. 
Using_VBG a_DT cell_NN line_NN that_WDT is_VBZ stably_RB transfected_VBN with_IN a_DT trimer_NN of_IN the_DT NFAT_* -1_NN regulatory_JJ element_NN linked_VBN to_TO a_DT lac-Z_NN reporter_NN gene_NN ,_, we_PRP screened_VBD for_IN inhibitors_NNS of_IN NFAT_* -1-mediated_JJ beta-galactosidase_NN activity_NN ._. 
WIN_NN 61058_CD and_CC WIN_NN 53071_CD were_VBD identified_VBN as_IN microM_NN inhibitors_NNS ._. 
These_DT compounds_NNS also_RB inhibited_VBD beta-galactosidase_NN mRNA_NN levels_NNS ._. 
Similar_JJ inhibition_NN of_IN NFAT_* -1-mediated_JJ gene_NN expression_NN was_VBD observed_VBN in_IN a_DT second_JJ cell_NN line_NN ,_, which_WDT is_VBZ stably_RB transfected_VBN with_IN NFAT_* -1_NN regulatory_JJ elements_NNS linked_VBN to_TO the_DT reporter_NN gene_NN for_IN sCD8_NN ._. 
At_IN 10_CD microM_NN ,_, both_DT compounds_NNS inhibited_VBD IL-2_NN mRNA_NN and_CC protein_NN levels_NNS in_IN the_DT NFAT_* -1-linked_JJ lac-Z_NN transfectants_NNS ,_, and_CC in_IN human_JJ lymphocytes_NNS ._. 
Both_DT compounds_NNS inhibited_VBD the_DT mixed_JJ lymphocyte_NN reaction_NN ,_, and_CC this_DT inhibition_NN was_VBD reversed_VBN by_IN exogenous_JJ IL-2_NN ._. 
WIN_NN 53071_CD inhibited_VBD IL-2_NN production_NN induced_VBN in_IN the_DT calcium-dependent_JJ PMA_NN and_CC ionomycin_NN pathway_NN ._. 
Conversely_RB ,_, calcium-independent_JJ anti-CD28_JJ Ab_NN and_CC PMA-induced_JJ IL-2_NN production_NN was_VBD resistant_JJ ._. 
Both_DT compounds_NNS altered_VBD the_DT NFAT_* -1_NN transcriptional_JJ complex_NN ,_, causing_VBG its_PRP$ retarded_VBN mobility_NN on_IN gels_NNS ._. 
By_IN these_DT functional_JJ criteria_NNS ,_, we_PRP believe_VBP we_PRP have_VBP identified_VBN two_CD structurally_RB distinct_JJ ,_, novel_JJ inhibitors_NNS of_IN NFAT_* -1-mediated_JJ transcription_NN ._. 
Transcriptional_JJ regulation_NN of_IN the_DT vacuolar_JJ H(+)-ATPase_NN B2_NN subunit_NN gene_NN in_IN differentiating_VBG THP-1_NN cells_NNS ._. 
Monocyte-macrophage_JJ differentiation_NN was_VBD used_VBN as_IN a_DT model_NN system_NN for_IN studying_VBG gene_NN regulation_NN of_IN the_DT human_JJ vacuolar_NN H(+)-ATPase_NN (_( V-ATPase_NN )_) ._. 
We_PRP examined_VBD mRNA_NN levels_NNS of_IN various_JJ V-ATPase_NN subunits_NNS during_IN differentiation_NN of_IN both_CC native_JJ monocytes_NNS and_CC the_DT cell_NN line_NN THP-1_NN ,_, and_CC found_VBD that_IN transcriptional_JJ and_CC post-transcriptional_JJ mechanisms_NNS could_MD account_VB for_IN increases_NNS in_IN cell_NN V-ATPase_NN content_NN ._. 
From_IN nuclear_JJ runoff_JJ experiments_NNS ,_, we_PRP found_VBD that_IN one_CD subunit_NN in_IN particular_JJ ,_, the_DT B2_NN isoform_NN (_( Mr_NN =_JJ 56,000_CD )_) ,_, was_VBD amplified_VBN primarily_RB by_IN transcriptional_JJ means_NNS ._. 
We_PRP have_VBP begun_VBN to_TO examine_VB the_DT structure_NN of_IN the_DT B2_NN subunit_NN promoter_NN region_NN ._. 
Isolation_NN and_CC sequencing_NN of_IN the_DT first_JJ exon_NN and_CC 5'-flanking_JJ region_NN of_IN this_DT gene_NN reveal_VBP a_DT TATA-less_JJ promoter_NN with_IN a_DT high_JJ G_NN +_CC C_NN content_NN ._. 
Primer_NN extension_NN and_CC ribonuclease_NN protection_NN analyses_NNS indicate_VBP a_DT single_JJ major_JJ transcriptional_JJ start_NN site_NN ._. 
We_PRP transfected_VBD promoter-luciferase_NN reporter_NN plasmids_NNS into_IN THP-1_NN cells_NNS to_TO define_VB sequences_NNS that_WDT mediate_VBP transcriptional_JJ control_NN during_IN monocyte_NN differentiation_NN ._. 
We_PRP found_VBD that_IN sequences_NNS downstream_RB from_IN the_DT transcriptional_JJ start_NN site_NN were_VBD sufficient_JJ to_TO confer_VB increased_VBN expression_NN during_IN THP-1_NN differentiation_NN ._. 
DNase_NN I_CD footprinting_NN and_CC sequence_NN analysis_NN revealed_VBD the_DT existence_NN of_IN multiple_JJ AP2_NN and_CC Sp1_NN binding_NN sites_NNS in_IN the_DT 5'-untranslated_JJ and_CC proximal_JJ coding_VBG regions_NNS ._. 
Synergy_NN between_IN signal_NN transduction_NN pathways_NNS is_VBZ obligatory_JJ for_IN expression_NN of_IN c-fos_NN in_IN B_NN and_CC T_NN cell_NN lines_NNS :_: implication_NN for_IN c-fos_NN control_NN via_IN surface_NN immunoglobulin_NN and_CC T_NN cell_NN antigen_NN receptors_NNS ._. 
Expression_NN of_IN the_DT protooncogene_NN c-fos_NN is_VBZ controlled_VBN by_IN three_CD main_JJ regulatory_JJ pathways_NNS involving_VBG kinase_NN C_NN ,_, cAMP_NN ,_, and_CC calcium_NN ._. 
Kinase_NN C_NN mediates_VBZ its_PRP$ effects_NNS via_IN phosphorylation_NN of_IN serum_NN response_NN factor_NN (_( SRF_NN )_) which_WDT interacts_VBZ with_IN the_DT serum_NN response_NN element_NN (_( SRE_NN )_) ;_: cAMP_NN and_CC calcium_NN mediate_VBP their_PRP$ effects_NNS via_IN phosphorylation_NN of_IN CREB_NN (_( cAMP_NN regulatory_JJ element_NN binding_NN protein_NN )_) presumably_RB by_IN activation_NN of_IN a_DT protein_NN kinase_NN A_NN or_CC calmodulin-regulated_JJ kinase_NN ._. 
We_PRP have_VBP examined_VBN the_DT function_NN of_IN these_DT elements_NNS in_IN Burkitt_NN 's_POS lymphoma_NN cells_NNS (_( Ramos_NN and_CC Daudi_NN )_) as_RB well_RB as_IN a_DT T_NN lymphocytic_JJ cell_NN line_NN (_( Jurkat_NN )_) ._. 
We_PRP have_VBP found_VBN that_IN stimulation_NN of_IN any_DT one_CD of_IN these_DT pathways_NNS alone_RB has_VBZ little_JJ or_CC no_DT effect_NN on_IN c-fos_NN induction_NN ._. 
However_RB ,_, kinase_NN C_NN activation_NN (_( PMA_NN stimulation_NN )_) combined_VBN with_IN either_CC cAMP_NN (_( forskolin_NN plus_CC MIX_NN )_) or_CC calcium_NN stimulation_NN (_( ionophore_NN )_) leads_VBZ to_TO greatly_RB enhanced_VBN c-fos_NN induction_NN ._. 
By_IN contrast_NN ,_, cAMP_NN in_IN the_DT presence_NN of_IN calcium_NN shows_VBZ no_DT synergy_NN in_IN c-fos_NN induction_NN ._. 
Okadaic_JJ acid_NN augments_VBZ PMA-_NN as_RB well_RB as_IN calcium-_* mediated_JJ activation_NN of_IN c-fos_NN ,_, and_CC has_VBZ little_JJ or_CC no_DT effect_NN when_WRB combined_VBN with_IN cAMP_NN ._. 
The_DT main_JJ difference_NN between_IN Ramos_NN (_( B_NN cells_NNS )_) and_CC Jurkat_NN (_( T_NN cells_NNS )_) in_IN the_DT regulation_NN of_IN c-fos_NN is_VBZ that_IN cAMP_NN plus_CC calcium_NN is_VBZ strongly_RB synergistic_JJ in_IN Jurkat_NN and_CC is_VBZ without_IN effect_NN in_IN Ramos_NN ._. 
Analysis_NN of_IN AP-1_NN activity_NN using_VBG gel_NN mobility_NN shift_NN assays_NNS confirms_VBZ that_IN the_DT requirements_NNS for_IN synergy_NN in_IN c-fos_NN mRNA_NN induction_NN are_VBP paralleled_VBN by_IN requirements_NNS for_IN synergy_NN in_IN induction_NN of_IN AP-1_NN activity_NN ._. 
Signaling_NN in_IN B_NN cells_NNS due_JJ to_TO anti-Ig_JJ stimulation_NN involves_VBZ both_CC kinase_NN C_NN activation_NN and_CC release_NN of_IN intracellular_JJ calcium_NN ,_, and_CC results_VBZ in_IN c-fos_NN mRNA_NN induction_NN ._. 
Our_PRP$ results_NNS indicate_VBP that_IN synergy_NN between_IN the_DT kinase_NN C_NN activation_NN and_CC calcium_NN is_VBZ needed_VBN for_IN efficient_JJ c-fos_NN induction_NN since_IN neither_DT of_IN these_DT two_CD alone_RB induces_VBZ c-fos_NN well_RB ._. 
That_IN synergy_NN of_IN signaling_NN pathways_NNS is_VBZ relevant_JJ for_IN the_DT anti-Ig_JJ induction_NN of_IN c-fos_NN is_VBZ supported_VBN by_IN the_DT fact_NN that_IN cAMP_* -inducing_JJ agents_NNS and_CC okadaic_JJ acid_NN further_RB enhance_VBP anti-Ig_JJ induction_NN of_IN c-fos_NN ._. 
These_DT results_NNS suggest_VBP that_IN cell-specific_JJ patterns_NNS of_IN synergy_NN are_VBP an_DT essential_JJ feature_NN for_IN c-fos_NN induction_NN and_CC may_MD be_VB relevant_JJ for_IN c-fos_NN control_NN through_IN B_NN and_CC T_NN cell_NN antigen_NN receptors_NNS ._. 
Multiple_JJ signals_NNS are_VBP required_VBN for_IN function_NN of_IN the_DT human_JJ granulocyte-macrophage_JJ colony-stimulating_JJ factor_NN gene_NN promoter_NN in_IN T_NN cells_NNS ._. 
The_DT human_JJ granulocyte-macrophage_JJ CSF_NN (_( GM-CSF_NN )_) gene_NN is_VBZ expressed_VBN in_IN T_NN cells_NNS in_IN response_NN to_TO TCR_NN activation_NN that_WDT can_MD be_VB mimicked_VBN by_IN treatment_NN of_IN the_DT cells_NNS with_IN PMA_NN and_CC Ca2+_NN ionophore_NN ._. 
The_DT gene_NN contains_VBZ a_DT proximal_JJ functional_JJ promoter_NN region_NN (_( -620_CD to_TO +34_CD )_) ,_, as_RB well_RB as_IN a_DT powerful_JJ enhancer_NN located_JJ 3_CD kb_NN upstream_RB ,_, both_DT of_IN which_WDT are_VBP involved_VBN in_IN the_DT response_NN of_IN the_DT gene_NN to_TO TCR_NN activation_NN ._. 
The_DT proximal_JJ promoter_NN contains_VBZ a_DT region_NN termed_VBN CLEO_NN (_( -54_CD to_TO -31_CD )_) that_WDT consists_VBZ of_IN a_DT purine_* -rich_JJ element_NN abutting_VBG an_DT activator_NN protein-1_NN (_( AP-1_NN )_) -like_JJ site_NN ,_, as_RB well_RB as_IN an_DT upstream_JJ nuclear_JJ factor-kappa_NN B_NN (_( NF-kappa_NN B_NN )_) site_NN (_( -85_CD to_TO -76_CD )_) and_CC a_DT CK-1_NN element_NN (_( -101_CD to_TO -92_CD )_) ._. 
We_PRP show_VBP in_IN this_DT work_NN that_IN mutations_NNS in_IN either_CC the_DT purine_* -rich_JJ region_NN of_IN the_DT CLEO_NN element_NN or_CC the_DT NF-kappa_NN B_NN site_NN result_VBP in_IN reduced_VBN PMA_* /_* Ca2+_NN activation_NN of_IN a_DT 620-bp_JJ human_JJ GM-CSF_NN promoter-_NN luciferase_NN reporter_NN construct_NN in_IN Jurkat_NN T_NN cells_NNS by_IN 65_CD %_NN and_CC 50_CD %_NN ,_, respectively_RB ._. 
The_DT major_JJ inducible_JJ protein_NN complex_NN that_WDT binds_VBZ to_TO the_DT human_JJ CLEO_NN (_( hCLEO_NN )_) element_NN is_VBZ an_DT AP-1_* -like_JJ complex_NN that_WDT is_VBZ inducible_JJ by_IN PMA_NN alone_RB ,_, but_CC shows_VBZ increased_VBN binding_NN in_IN response_NN to_TO PMA_NN together_RB with_IN Ca2+_NN ionophore_NN ._. 
Although_IN the_DT binding_NN of_IN this_DT complex_NN is_VBZ not_RB cyclosporin_* -sensitive_JJ ,_, promoter_NN induction_NN is_VBZ inhibited_VBN by_IN cyclosporin_NN treatment_NN ._. 
A_DT second_JJ weak_JJ inducible_JJ complex_NN resembling_VBG nuclear_JJ factor_NN of_IN activated_VBN T_NN cells_NNS (_( NF-AT_NN )_) was_VBD also_RB observed_VBN binding_VBG to_TO the_DT hCLEO_NN region_NN ._. 
By_IN using_VBG recombinant_JJ proteins_NNS ,_, we_PRP confirmed_VBD that_IN AP-1_NN ,_, NF-ATp_NN ,_, and_CC a_DT higher_JJR order_NN NF-ATp_NN /_* AP-1_NN complex_NN could_MD all_DT form_VB with_IN the_DT hCLEO_NN element_NN ,_, and_CC we_PRP have_VBP also_RB defined_VBN the_DT sequence_NN requirements_NNS for_IN binding_NN of_IN each_DT of_IN these_DT complexes_NNS ._. 
We_PRP found_VBD that_IN expression_NN of_IN a_DT constitutively_RB active_JJ form_NN of_IN calcineurin_NN could_MD substitute_VB for_IN Ca2+_NN ionophore_NN and_CC synergize_VB with_IN PMA_NN to_TO activate_VB the_DT GM-CSF_NN promoter_NN ,_, and_CC conversely_RB that_IN mutant-activated_JJ Ras_NN could_MD substitute_VB for_IN PMA_NN and_CC cooperate_VB with_IN Ca2+_NN ionophore_NN ._. 
Co-expression_NN of_IN Ras_NN and_CC calcineurin_NN ,_, however_RB ,_, did_VBD not_RB activate_VB the_DT GM-CSF_NN promoter_NN ,_, but_CC required_VBD the_DT additional_JJ expression_NN of_IN NF-kappa_NN B_NN p65_NN ._. 
These_DT results_NNS imply_VBP that_IN at_IN least_JJS three_CD signals_NNS are_VBP required_VBN to_TO activate_VB the_DT GM-CSF_NN proximal_JJ promoter_NN ,_, and_CC that_IN the_DT signals_NNS impinge_VBP on_IN distinct_JJ transcription_NN factors_NNS that_WDT bind_VBP to_TO the_DT hCLEO_NN and_CC NF-kappa_NN B_NN regions_NNS of_IN the_DT promoter_NN ._. 
IL-10_NN induces_VBZ the_DT tyrosine_NN phosphorylation_NN of_IN tyk2_NN and_CC Jak1_NN and_CC the_DT differential_JJ assembly_NN of_IN STAT1_NN alpha_NN and_CC STAT3_NN complexes_NNS in_IN human_JJ T_NN cells_NNS and_CC monocytes_NNS ._. 
IL-10_NN affects_VBZ monocytes_NNS and_CC T_NN cells_NNS by_IN driving_VBG the_DT progression_NN of_IN immune_JJ responsiveness_NN such_IN that_IN Th2_NN lymphocyte-mediated_JJ effects_NNS predominate_VBP ._. 
In_IN this_DT report_NN ,_, we_PRP show_VBP that_IN in_IN monocytes_NNS and_CC T_NN cells_NNS IL-10_NN stimulates_VBZ tyrosine_NN phosphorylation_NN of_IN the_DT signal_NN transducers_NNS and_CC activators_NNS of_IN transcription_NN ,_, STAT1_NN alpha_NN and_CC STAT3_NN ,_, in_IN a_DT differential_JJ manner_NN such_IN that_IN the_DT relative_JJ formation_NN of_IN homo-_JJ and_CC hetero_* dimers_NNS varies_VBZ between_IN the_DT two_CD cell_NN types_NNS ._. 
Moreover_RB ,_, monocytes_NNS express_VBP a_DT novel_JJ IL-10_* -stimulated_JJ STAT_NN protein_NN with_IN an_DT M(r)_NN of_IN 70_CD kDa_NN that_WDT is_VBZ recognized_VBN by_IN the_DT anti-_* STAT3_JJ Ab_NN but_CC is_VBZ not_RB observed_VBN in_IN T_NN cells_NNS ._. 
IL-10_NN treatment_NN of_IN both_CC T_NN cells_NNS and_CC monocytes_NNS results_VBZ in_IN the_DT ligand-induced_JJ tyrosine_NN phosphorylation_NN of_IN tyk2_NN and_CC Jak1_NN ,_, but_CC not_RB Jak2_NN or_CC Jak3_NN ._. 
Selective_JJ modulation_NN of_IN immune_JJ responsiveness_NN by_IN IL-10_NN in_IN cells_NNS such_JJ as_IN monocytes_NNS and_CC T_NN cells_NNS may_MD result_VB in_IN part_NN from_IN the_DT differential_JJ activation_NN of_IN STAT_NN protein_NN pairs_NNS ._. 
Use_NN of_IN new_JJ biologic_JJ markers_NNS in_IN the_DT ovulation_NN induction_NN ._. 
Biological_JJ markers_NNS of_IN ovulation_NN ,_, after_IN a_DT great_JJ in_IN the_DT past_NN ,_, have_VBP been_VBN fallen_VBN into_IN disuse_NN for_IN the_DT large_JJ diffusion_NN of_IN biochemical_JJ and_CC biophysical_JJ ones_NNS ._. 
However_RB ,_, the_DT real_JJ effect_NN of_IN hormones_NNS involved_VBN in_IN ovulation_NN is_VBZ expressed_VBN by_IN biological_JJ modifications_NNS on_IN target_NN tissues_NNS ._. 
To_TO explore_VB the_DT modifications_NNS of_IN not_RB reproductive_JJ target_NN tissues_NNS as_IN ovulation_NN markers_NNS we_PRP studied_VBD the_DT behaviour_NN of_IN Albuminemia_NN ,_, Platelet_NN Factor_NN IV_CD (_( as_IN indicator_NN of_IN Platelet_NN Aggregation_NN )_) ,_, Type_NN II_CD estrogenic_JJ receptors_NNS in_IN 42_CD ovulation_NN induced_JJ women_NNS ,_, undergoing_VBG our_PRP$ observation_NN ._. 
33_CD of_IN them_PRP had_VBD ovulation_NN and_CC 9_CD developed_VBD a_DT LUF_NN syndrome_NN ,_, constituting_VBG two_CD biological_JJ models_NNS of_IN an_DT opposite_JJ situation_NN for_IN the_DT three_CD markers_NNS observed_VBN ._. 
All_PDT the_DT markers_NNS considered_VBN were_VBD sufficiently_RB sensitive_JJ ,_, but_CC among_IN them_PRP ,_, Platelet_NN Factor_NN IV_CD was_VBD the_DT most_RBS reliable_JJ to_TO the_DT hormonal_JJ ovulatory_JJ situation_NN ._. 
Abnormal_JJ regulation_NN of_IN the_DT IL-2_NN promoter_NN in_IN lpr_NN lymphocytes_NNS results_VBZ in_IN constitutive_JJ expression_NN of_IN a_DT novel_JJ nuclear_JJ factor_NN of_IN activated_VBN T_NN cells_* -binding_JJ factor_NN ._. 
The_DT inert_JJ quality_NN of_IN MRL-Ipr/Ipr_NN (_( Ipr_NN )_) peripheral_JJ CD4-CD8-_JJ (_( CD4-8-_NN )_) T_NN cells_NNS manifests_VBZ primarily_RB as_IN an_DT inability_NN to_TO proliferate_VB or_CC produce_VB IL-2_NN in_IN response_NN to_TO TCR_NN or_CC mitogenic_JJ stimulation_NN ._. 
Yet_RB these_DT same_JJ cells_NNS do_VBP initiate_VB early_JJ TCR_* -mediated_JJ signaling_NN events_NNS ,_, such_JJ as_IN generation_NN of_IN inositol_JJ phosphates_NNS and_CC increased_VBN intracellular_JJ calcium_NN ._. 
They_PRP also_RB display_VBP constitutively_RB high_JJ levels_NNS of_IN p59fyn_NN and_CC CD3_NN zeta_NN tyrosine_NN phosphorylation_NN ._. 
The_DT generation_NN of_IN second_JJ messengers_NNS in_IN T_NN cells_NNS normally_RB leads_VBZ to_TO downstream_JJ signaling_NN that_WDT results_VBZ in_IN transcriptional_JJ activation_NN of_IN the_DT IL-2_NN gene_NN ._.