Response to intranasal fluticasone propionate in perennial allergic rhinitis not associated with glucocorticoid receptor characteristics .

Response to intranasal fluticasone propionate in perennial allergic rhinitis not associated with <protein>glucocorticoid receptor</protein> characteristics .


BACKGROUND : The reduction of symptoms due to treatment with corticosteroids varies among patients with perennial rhinitis .


Most patients will respond but a few patients respond less to these drugs .


OBJECTIVE : To investigate the association in reduction of symptoms due to glucocorticoids and glucocorticoid receptor characteristics in patients with perennial allergic rhinitis , in vitro glucocorticoid receptor binding studies were performed with peripheral blood mononuclear cells using dexamethasone and in vitro production of mediators were measured .

OBJECTIVE : To investigate the association in reduction of symptoms due to glucocorticoids and <protein>glucocorticoid receptor</protein> characteristics in patients with perennial allergic rhinitis , in vitro <protein>glucocorticoid receptor</protein> binding studies were performed with <cell_type>peripheral blood mononuclear cells</cell_type> using dexamethasone and in vitro production of <protein>mediators</protein> were measured .


METHODS : During a double-blind placebo-controlled crossover study , 200 micrograms fluticasone propionate aqueous nasal spray ( in the active treatment period ) and placebo ( in the placebo treatment period ) were administered twice daily for 2 weeks to 22 patients allergic to house dust mite .


At the end of both treatment periods symptoms were scored after allergen provocation ( 100 , 1000 , 10000 BU/mL ) and during the 9.5 hours after this challenge .


Receptor binding studies with dexamethasone were performed with peripheral blood mononuclear cells .

Receptor binding studies with dexamethasone were performed with <cell_type>peripheral blood mononuclear cells</cell_type> .


Leukotriene B4 produced by monocytes in vitro and soluble interleukin-2 receptor released by lymphocytes in vitro and cortisol levels in plasma were determined .

Leukotriene B4 produced by <cell_type>monocytes</cell_type> in vitro and soluble <protein>interleukin-2 receptor</protein> released by <cell_type>lymphocytes</cell_type> in vitro and cortisol levels in plasma were determined .


RESULTS : No significant partial correlations of the number of the peripheral blood mononuclear cell glucocorticoid receptors ( 6821 +/- 5669 binding sites per cell ) and the affinity ( Kd : 16.5 +/- 13.51 nmol/L ) for the glucocorticoid receptors with the symptom score ( placebo : 4.3 +/- 2.45 pts ; fluticasone : 2.4 +/- 1.55 pts ) after active treatment were found .

RESULTS : No significant partial correlations of the number of the <protein>peripheral blood mononuclear cell glucocorticoid receptors</protein> ( 6821 +/- 5669 binding sites per cell ) and the affinity ( Kd : 16.5 +/- 13.51 nmol/L ) for the <protein>glucocorticoid receptors</protein> with the symptom score ( placebo : 4.3 +/- 2.45 pts ; fluticasone : 2.4 +/- 1.55 pts ) after active treatment were found .


Also no significant partial correlations of the levels of leukotriene B4 ( 45.6 +/- 105.3 ng/10 ( 6 ) cells ) produced by monocytes in vitro , soluble interleukin-2 receptor ( 734 +/- 237 ng/10 ( 6 ) cells ) released by lymphocytes in vitro and cortisol levels ( 571 +/- 236 ng/mL ) in plasma with the symptom score after active treatment were found .

Also no significant partial correlations of the levels of leukotriene B4 ( 45.6 +/- 105.3 ng/10 ( 6 ) cells ) produced by <cell_type>monocytes</cell_type> in vitro , soluble <protein>interleukin-2 receptor</protein> ( 734 +/- 237 ng/10 ( 6 ) cells ) released by <cell_type>lymphocytes</cell_type> in vitro and cortisol levels ( 571 +/- 236 ng/mL ) in plasma with the symptom score after active treatment were found .


CONCLUSIONS : The reduction of symptoms due to topical fluticasone propionate in patients with rhinitis and allergy to house dust mite is not correlated with the characteristics of the glucocorticoid receptor .

CONCLUSIONS : The reduction of symptoms due to topical fluticasone propionate in patients with rhinitis and allergy to house dust mite is not correlated with the characteristics of the <protein>glucocorticoid receptor</protein> .


Constitutive expression of p50 homodimer in freshly isolated human monocytes decreases with in vitro and in vivo differentiation : a possible mechanism influencing human immunodeficiency virus replication in monocytes and mature macrophages .

Constitutive expression of <protein>p50 homodimer</protein> in <cell_type>freshly isolated human monocytes</cell_type> decreases with in vitro and in vivo differentiation : a possible mechanism influencing human immunodeficiency virus replication in <cell_type>monocytes</cell_type> and mature macrophages .


Human immunodeficiency virus type 1 ( HIV-1 ) replicates more efficiently in vitro in differentiated macrophages than in freshly isolated monocytes .

Human immunodeficiency virus type 1 ( HIV-1 ) replicates more efficiently in vitro in <cell_type>differentiated macrophages</cell_type> than in <cell_type>freshly isolated monocytes</cell_type> .


We investigated whether this may be partly explained by changes in expression of NF-kappaB with monocyte differentiation .

We investigated whether this may be partly explained by changes in expression of <protein>NF-kappaB</protein> with monocyte differentiation .


We demonstrated that constitutive expression of NF-kappaB in primary human monocytes changed significantly with differentiation in vitro to monocyte-derived macrophages ( MDMs ) and differentiation in vivo to alveolar macrophages ( AMs ) .

We demonstrated that constitutive expression of <protein>NF-kappaB</protein> in <cell_type>primary human monocytes</cell_type> changed significantly with differentiation in vitro to <cell_type>monocyte-derived macrophages</cell_type> ( <cell_type>MDMs</cell_type> ) and differentiation in vivo to <cell_type>alveolar macrophages</cell_type> ( <cell_type>AMs</cell_type> ) .


Freshly isolated monocytes constitutively expressed high levels of transcriptionally inactive p50 homodimer which decreased with time in culture in favor of the transcriptionally active p50/p65 and p50/RelB heterodimers .

Freshly isolated monocytes constitutively expressed high levels of <protein>transcriptionally inactive p50 homodimer</protein> which decreased with time in culture in favor of the transcriptionally active <protein>p50/p65 and p50/RelB heterodimers</protein> .


As in MDMs , AMs constitutively expressed p50/p65 and p50/RelB although at lower levels .

As in <cell_type>MDMs</cell_type> , <cell_type>AMs</cell_type> constitutively expressed <protein>p50/p65</protein> and <protein>p50/RelB</protein> although at lower levels .


HIV infection of fresh monocytes failed to induce p50/p65 as seen in MDMs .

HIV infection of fresh <cell_type>monocytes</cell_type> failed to induce <protein>p50/p65</protein> as seen in <cell_type>MDMs</cell_type> .


The replacement of p50 homodimers with transcriptionally active heterodimers following time in culture may partially explain the progressive increase in susceptibility of monocytes to HIV infection during in vitro culture .

The replacement of <protein>p50 homodimers</protein> with transcriptionally active heterodimers following time in culture may partially explain the progressive increase in susceptibility of <cell_type>monocytes</cell_type> to HIV infection during in vitro culture .


The change in NF-kappaB components with monocyte differentiation in vivo may also explain the different transcriptional activities of these cell populations in HIV-infected individuals .

The change in <protein>NF-kappaB components</protein> with monocyte differentiation in vivo may also explain the different transcriptional activities of these cell populations in HIV-infected individuals .


NF-kappa B -independent suppression of HIV expression by ascorbic acid .

<protein>NF-kappa B</protein> -independent suppression of HIV expression by ascorbic acid .


Ascorbic acid ( ascorbate or vitamin C ) has been shown to suppress the induction of HIV in latently infected T lymphocytic cells following stimulation with a tumor promoter ( PMA ) and inflammatory cytokine ( TNF-alpha ) .

Ascorbic acid ( ascorbate or vitamin C ) has been shown to suppress the induction of HIV in <cell_type>latently infected T lymphocytic cells</cell_type> following stimulation with a tumor promoter ( PMA ) and <protein>inflammatory cytokine</protein> ( <protein>TNF-alpha</protein> ) .


To assess whether this inhibition was mediated via modulation of the cellular transcription factor , NF-kappa B , we carried out gel shift analysis on nuclear extracts prepared under different conditions of cell stimulation in the presence or absence of ascorbate , N-acetylcysteine ( NAC ) , or zidovudine ( AZT ) .

To assess whether this inhibition was mediated via modulation of the <protein>cellular transcription factor</protein> , <protein>NF-kappa B</protein> , we carried out gel shift analysis on nuclear extracts prepared under different conditions of cell stimulation in the presence or absence of ascorbate , N-acetylcysteine ( NAC ) , or zidovudine ( AZT ) .


Pretreatment of ACH-2 T cells by NAC followed by stimulation with PMA , TNF-alpha , or hydrogen peroxide ( H2O2 ) resulted in strong suppression of NF-kappa B activation .

Pretreatment of <cell_line>ACH-2 T cells</cell_line> by NAC followed by stimulation with PMA , <protein>TNF-alpha</protein> , or hydrogen peroxide ( H2O2 ) resulted in strong suppression of <protein>NF-kappa B</protein> activation .


In contrast , neither ascorbate nor AZT affected NF-kappa B activity under all three induction conditions in the ACH-2 cell line .

In contrast , neither ascorbate nor AZT affected <protein>NF-kappa B</protein> activity under all three induction conditions in the <cell_line>ACH-2 cell line</cell_line> .


Ascorbate and AZT also had no effect on NF-kappa B activation following TNF-alpha- or PMA-induced stimulation of U1 promonocytic cells .

Ascorbate and AZT also had no effect on <protein>NF-kappa B</protein> activation following TNF-alpha- or PMA-induced stimulation of U1 promonocytic cells .


These results suggest that the molecular mechanism of HIV inhibition by ascorbate is not mediated via NF-kappa B inhibition , unlike that seen with other antioxidants .

These results suggest that the molecular mechanism of HIV inhibition by ascorbate is not mediated via <protein>NF-kappa B</protein> inhibition , unlike that seen with other antioxidants .


A T cell-specific enhancer in the interleukin-3 locus is activated cooperatively by Oct and NFAT elements within a DNase I-hypersensitive site .

A <dna>T cell-specific enhancer</dna> in the <dna>interleukin-3 locus</dna> is activated cooperatively by <dna>Oct and NFAT elements</dna> within a <dna>DNase I-hypersensitive site</dna> .


Interleukin-3 ( IL-3 ) is a cytokine that is expressed primarily in activated T cells .

<protein>Interleukin-3</protein> ( <protein>IL-3</protein> ) is a <protein>cytokine</protein> that is expressed primarily in <cell_type>activated T cells</cell_type> .


Here we identified an inducible T cell-specific enhancer 14 kb upstream of the IL-3 gene that responded to activation of T cell receptor signaling pathways .

Here we identified an inducible <dna>T cell-specific enhancer</dna> 14 kb upstream of the <dna>IL-3 gene</dna> that responded to activation of <protein>T cell receptor</protein> signaling pathways .


The IL-3 enhancer spanned an inducible cyclosporin A-sensitive DNase I-hypersensitive site found only in T cells .

The <protein>IL-3</protein> enhancer spanned an inducible <dna>cyclosporin A-sensitive DNase I-hypersensitive site</dna> found only in <cell_type>T cells</cell_type> .


Four NFAT-like elements exist within the enhancer .

Four <dna>NFAT-like elements</dna> exist within the enhancer .


The two most active NFAT-like elements were located at the center of the DNase I-hypersensitive site .

The two most active <dna>NFAT-like elements</dna> were located at the center of the <dna>DNase I-hypersensitive site</dna> .


One of these NFAT-like elements encompassed overlapping Oct- and NFATp/c-binding sites , which functioned in a highly synergistic manner .

One of these <dna>NFAT-like elements</dna> encompassed overlapping <dna>Oct- and NFATp/c-binding sites</dna> , which functioned in a highly synergistic manner .


We suggest that the T cell-specific expression of the IL-3 gene is partly controlled through the enhancer by cooperation between Oct and NFAT family proteins .

We suggest that the T cell-specific expression of the <dna>IL-3 gene</dna> is partly controlled through the enhancer by cooperation between <protein>Oct and NFAT family proteins</protein> .


Adenovirus E1B 19K protein is required for efficient DNA replication in U937 cells .

<protein>Adenovirus E1B 19K protein</protein> is required for efficient DNA replication in <cell_line>U937 cells</cell_line> .


The adenovirus E1B 19K gene plays an essential role in transformation of primary rodent cells in cooperation with E1A and in the inhibition of apoptosis during lytic infection .

The <dna>adenovirus E1B 19K gene</dna> plays an essential role in transformation of <cell_type>primary rodent cells</cell_type> in cooperation with <protein>E1A</protein> and in the inhibition of apoptosis during lytic infection .


It has been shown that this E1B 19K protein is not necessary for viral DNA replication in human cell lines , such as HeLa and KB .

It has been shown that this <protein>E1B 19K protein</protein> is not necessary for viral DNA replication in <cell_line>human cell lines</cell_line> , such as <cell_line>HeLa</cell_line> and <cell_line>KB</cell_line> .


We reported here that the E1B 19K mutant viruses were unable to replicate efficiently in a monocyte cell line , U937 .

We reported here that the <protein>E1B 19K</protein> mutant viruses were unable to replicate efficiently in a <cell_line>monocyte cell line</cell_line> , <cell_line>U937</cell_line> .


Viral DNA synthesis and late gene expression were found to be defective in U937 cells infected with E1B 19K mutants compared with wild-type virus .

Viral DNA synthesis and late gene expression were found to be defective in <cell_line>U937 cells</cell_line> infected with <protein>E1B 19K</protein> mutants compared with wild-type virus .


Early viral RNA splicing patterns also differ between wild-type and dl337-infected cells .

Early viral RNA splicing patterns also differ between <cell_line>wild-type and dl337-infected cells</cell_line> .


Furthermore , the defect in viral replication could be complemented by dl312 virus defective in E1A expression 4 days after infection with E1B mutants , suggesting persistence of the E1B mutant genome in the infected cells despite defective onset of the late phase of replication .

Furthermore , the defect in viral replication could be complemented by dl312 virus defective in <protein>E1A</protein> expression 4 days after infection with E1B mutants , suggesting persistence of the <dna>E1B mutant genome</dna> in the infected cells despite defective onset of the late phase of replication .


These results imply that E1B 19K is required for efficient viral DNA replication in U937 cells .

These results imply that <protein>E1B 19K</protein> is required for efficient viral DNA replication in <cell_line>U937 cells</cell_line> .


Inefficient DNA replication is also found in another monocyte cell line , THP-1

Inefficient DNA replication is also found in another <cell_line>monocyte cell line</cell_line> , <cell_line>THP-1</cell_line>


Activation of Ras and mitogen-activated protein kinase pathway by terminal complement complexes is G protein dependent .

Activation of Ras and <protein>mitogen-activated protein kinase</protein> pathway by <protein>terminal complement complexes</protein> is <protein>G protein</protein> dependent .


Assembly of terminal complement complexes ( TCC ) C5b-7 , C5b-8 , and C5b-9 on target cells during acute and chronic inflammation induces hydrolysis of plasma membrane phospholipids and heterotrimeric G protein activation .

Assembly of <protein>terminal complement complexes</protein> ( <protein>TCC</protein> ) <protein>C5b-7</protein> , <protein>C5b-8</protein> , and <protein>C5b-9</protein> on target cells during acute and chronic inflammation induces hydrolysis of plasma membrane phospholipids and heterotrimeric <protein>G protein</protein> activation .


TCC also stimulate a variety of cellular activities , which include cytokine synthesis , proto-oncogene activation , and mitotic signaling .

<protein>TCC</protein> also stimulate a variety of cellular activities , which include <protein>cytokine</protein> synthesis , <dna>proto-oncogene</dna> activation , and mitotic signaling .


Now we report that sublytic TCC induced Ras , Raf-1 , and extracellular signal-regulated kinase ( ERK ) 1 activation in JY25 B cell line .

Now we report that sublytic <protein>TCC</protein> induced <protein>Ras</protein> , <protein>Raf-1</protein> , and <protein>extracellular signal-regulated kinase ( ERK ) 1</protein> activation in JY25 B cell line .


When cells were exposed to C5b-9 , GTP-bound Ras in anti-C5b-9 immunoprecipitates was increased 3.2-fold at 2 min , while GTP-bound Ras in anti-Ras immunoprecipitates was increased 2-fold at 10 min .

When cells were exposed to <protein>C5b-9</protein> , <protein>GTP-bound Ras</protein> in <protein>anti-C5b-9 immunoprecipitates</protein> was increased 3.2-fold at 2 min , while <protein>GTP-bound Ras</protein> in <protein>anti-Ras immunoprecipitates</protein> was increased 2-fold at 10 min .


Both C5b-9 and C5b-7 , but not C5b6 , increased Raf-1 kinase activity maximum 3.3-fold at 2 min and 2.8-fold at 5 min , respectively .

Both <protein>C5b-9</protein> and <protein>C5b-7</protein> , but not <protein>C5b6</protein> , increased <protein>Raf-1</protein> kinase activity maximum 3.3-fold at 2 min and 2.8-fold at 5 min , respectively .


ERK1 activity was 2-fold increased by C5b-9 at 2 min and by C5b-7 at 10 min , over the C5b6 level .

<protein>ERK1</protein> activity was 2-fold increased by <protein>C5b-9</protein> at 2 min and by <protein>C5b-7</protein> at 10 min , over the <protein>C5b6</protein> level .


The role of mitogen-activated protein kinase ( MAPK ) pathway on TCC -inducible mitotic signaling was evaluated by assessing DNA synthesis and activator protein 1 ( AP-1 ) DNA-binding activity .

The role of <protein>mitogen-activated protein kinase</protein> ( <protein>MAPK</protein> ) pathway on <protein>TCC</protein> -inducible mitotic signaling was evaluated by assessing DNA synthesis and <protein>activator protein 1</protein> ( <protein>AP-1</protein> ) DNA-binding activity .


The MAPK/ERK-specific inhibitor PD 098 , 059 abolished the C5b-9 -induced DNA synthesis .

The MAPK/ERK-specific inhibitor PD 098 , 059 abolished the <protein>C5b-9</protein> -induced DNA synthesis .


Involvement of G protein in the activation of MAPK pathway by TCC was indicated by inhibition of Raf-1 and ERK1 kinase activity , as well as the DNA synthesis by pretreatment of cells with pertussis toxin .

Involvement of <protein>G protein</protein> in the activation of <protein>MAPK</protein> pathway by <protein>TCC</protein> was indicated by inhibition of <protein>Raf-1</protein> and <protein>ERK1</protein> kinase activity , as well as the DNA synthesis by pretreatment of cells with <protein>pertussis toxin</protein> .


Overexpression of beta-adrenergic receptor kinase 1 carboxyl-terminal peptide in JY25 cells also inhibited Raf-1 and ERK1 activity , indicating a direct involvement of G betagamma subunits in the signal transduction generated through activation of MAPK pathway by TCC assembly in the plasma membrane .

Overexpression of beta-adrenergic receptor kinase 1 carboxyl-terminal peptide in <cell_line>JY25 cells</cell_line> also inhibited <protein>Raf-1</protein> and <protein>ERK1</protein> activity , indicating a direct involvement of G betagamma subunits in the signal transduction generated through activation of <protein>MAPK</protein> pathway by <protein>TCC</protein> assembly in the plasma membrane .


Spontaneous occurrence of early region 1A reiteration mutants of type 5 adenovirus in persistently infected human T-lymphocytes .

Spontaneous occurrence of early region 1A reiteration mutants of type 5 adenovirus in <cell_type>persistently infected human T-lymphocytes</cell_type> .


Mutants of type 5 adenovirus ( Ad5 ) with reiterated DNA sequences in the E1a region appeared in a human T-lymphocyte cell line , Molt-4 , persistently infected with H5sub304 , a deletion/substitution mutant that has a wild-type phenotype in viral replication .

Mutants of type 5 adenovirus ( Ad5 ) with <dna>reiterated DNA sequences</dna> in the <dna>E1a region</dna> appeared in a <cell_line>human T-lymphocyte cell line</cell_line> , <cell_line>Molt-4</cell_line> , persistently infected with H5sub304 , a deletion/substitution mutant that has a wild-type phenotype in viral replication .


Endonuclease analyses and DNA sequencing revealed DNA reiteration in each mutant .

<protein>Endonuclease</protein> analyses and DNA sequencing revealed DNA reiteration in each mutant .


In the four representative mutants investigated , the DNA reiterations all started within a six-base-pair consensus sequence , G ( or C ) CTGTG , located in the second exon of the E1a region ( at nt 1333 , 1367 , or 1419 ) .

In the four representative mutants investigated , the <dna>DNA reiterations</dna> all started within a <dna>six-base-pair consensus sequence</dna> , G ( or C ) CTGTG , located in the second <dna>exon</dna> of the <dna>E1a region</dna> ( at nt 1333 , 1367 , or 1419 ) .


There was not any DNA homology between the breakpoints in the second exon and the inserting sequences ( starting at nt 532 , 710 , or 792 ) .

There was not any DNA homology between the breakpoints in the second <dna>exon</dna> and the <dna>inserting sequences</dna> ( starting at nt 532 , 710 , or 792 ) .


Northern analyses suggested that the reiterated splicing sites of the representative mutants were all used in RNA splicing , and the closest donor and recipient joints were used most frequently .

Northern analyses suggested that the <dna>reiterated splicing sites</dna> of the representative mutants were all used in RNA splicing , and the closest donor and recipient joints were used most frequently .


These observations imply that during persistent infection Ad5 underwent spontaneous mutations by sequence-specific breakage and nonhomologous end-end joining recombination events .


These E1a reiteration mutants could be propagated in HeLa , A549 , and KB cells ; they were genetically stable ; and they killed CREF cells at a strikingly high frequency .

These E1a reiteration mutants could be propagated in <cell_line>HeLa</cell_line> , <cell_line>A549</cell_line> , and <cell_line>KB cells</cell_line> ; they were genetically stable ; and they killed <cell_line>CREF cells</cell_line> at a strikingly high frequency .


Preliminary observations tend to correlate this CREF cell killing with the accumulation of the early viral proteins and/or viral DNA in the infected cells .

Preliminary observations tend to correlate this CREF cell killing with the accumulation of the <protein>early viral proteins</protein> and/or <dna>viral DNA</dna> in the <cell_type>infected cells</cell_type> .


This degree of cell damage was not observed in Ad5wt or H5sub304 infection of CREF cells .

This degree of cell damage was not observed in Ad5wt or H5sub304 infection of <cell_line>CREF cells</cell_line> .


The observed E1a reiterations provide a model to gain insight into understanding the evolutionary events of some , if not all , adenovirus types during many years of symbiotic , persistent relationship in human tonsils and adenoids and possibly other lymphoid organs .

The observed <dna>E1a reiterations</dna> provide a model to gain insight into understanding the evolutionary events of some , if not all , adenovirus types during many years of symbiotic , persistent relationship in human tonsils and adenoids and possibly other lymphoid organs .


Preferential presentation of herpes simplex virus T-cell antigen by HLA DQA1*0501/DQB1*0201 in comparison to HLA DQA1*0201/DQB1*0201 .

Preferential presentation of <protein>herpes simplex virus T-cell antigen</protein> by <protein>HLA DQA1*0501/DQB1*0201</protein> in comparison to <protein>HLA DQA1*0201/DQB1*0201</protein> .


The HLA DQA1 locus is polymorphic .

The <dna>HLA DQA1 locus</dna> is polymorphic .


Haplotypes containing HLA DQA1*0501 , but not HLA DQA1*0201 , together with HLA DQB1*0201 are associated with Grave 's disease and celiac sprue .

Haplotypes containing <dna>HLA DQA1*0501</dna> , but not <dna>HLA DQA1*0201</dna> , together with <dna>HLA DQB1*0201</dna> are associated with Grave 's disease and celiac sprue .


In this report , we demonstrate a functional correlate of DQA1 polymorphism .


T cells infiltrating a herpes simplex virus ( HSV ) lesion from a HLA DQ 2 , 7 individual yielded a virus-specific CD4+ clone restricted by DQ2 .

<cell_type>T cells</cell_type> infiltrating a herpes simplex virus ( HSV ) lesion from a HLA DQ 2 , 7 individual yielded a <cell_line>virus-specific CD4+ clone</cell_line> restricted by <protein>DQ2</protein> .


Presentation of viral peptide and protein segregated with DQA1 allele , because cell lines bearing DQA1*0501/DQB1*0201 heterodimers presented antigen in proliferation and cytotoxicity assays much more efficiently than cell lines bearing DQA1*0201/DQB1*0201 .

Presentation of viral peptide and protein segregated with <dna>DQA1 allele</dna> , because cell lines bearing <protein>DQA1*0501/DQB1*0201 heterodimers</protein> presented antigen in proliferation and cytotoxicity assays much more efficiently than <cell_line>cell lines</cell_line> bearing <protein>DQA1*0201/DQB1*0201</protein> .


Binding of viral peptide to cell lines bearing DQA1*0201 , in comparison to DQA1*0501 , was only moderately reduced and may not explain this effect .

Binding of viral peptide to cell lines bearing <protein>DQA1*0201</protein> , in comparison to <protein>DQA1*0501</protein> , was only moderately reduced and may not explain this effect .


Truncation and substitution analyses of peptide binding and T-cell activation were performed to determine which viral peptide residues contacting TCR might therefore be presented in an altered conformation by DQA1*0201/DQB1*0201 .

Truncation and substitution analyses of peptide binding and T-cell activation were performed to determine which viral peptide residues contacting <protein>TCR</protein> might therefore be presented in an altered conformation by <protein>DQA1*0201/DQB1*0201</protein> .


Residues 432 , 435 , 437 , 438 , and 440 ( position P1 , P4 , P6 , P7 , and P9 ) contributed to DQ2 binding , whereas residues 431 , 433 , 434 , and 436 ( positions P 1 , P2 , P3 , and P5 ) contributed to TCR contact .

Residues 432 , 435 , 437 , 438 , and 440 ( <dna>position P1 , P4 , P6 , P7 , and P9</dna> ) contributed to <protein>DQ2</protein> binding , whereas residues 431 , 433 , 434 , and 436 ( <dna>positions P 1 , P2 , P3 , and P5</dna> ) contributed to <protein>TCR</protein> contact .


Differential presentation of peptide by HLA DQ2 heterodimers varying at the DQA1 locus may have relevance to host defense and the pathogenesis of HLA DQ2 -associated autoimmune diseases .

Differential presentation of peptide by <protein>HLA DQ2 heterodimers</protein> varying at the <dna>DQA1 locus</dna> may have relevance to host defense and the pathogenesis of HLA <protein>DQ2</protein> -associated autoimmune diseases .


Inhibitor ( IK ) of IFN-gamma induced HLA class II antigens expression also inhibits HLA class II constitutive expression in the human Raji B cell line .

Inhibitor ( IK ) of <protein>IFN-gamma</protein> induced <protein>HLA class II antigens</protein> expression also inhibits <protein>HLA class II</protein> constitutive expression in the <cell_line>human Raji B cell line</cell_line> .


The expression of major histocompatibility complex ( MHC ) class II antigens is constitutive in professional antigen presenting cells ( APCs ) but can also be induced by interferon-gamma ( IFN-gamma ) on the majority of the non professional APCs ( e.g . fibroblasts ) .

The expression of <protein>major histocompatibility complex ( MHC ) class II antigens</protein> is constitutive in <cell_type>professional antigen presenting cells</cell_type> ( <cell_type>APCs</cell_type> ) but can also be induced by <protein>interferon-gamma</protein> ( <protein>IFN-gamma</protein> ) on the majority of the non <cell_type>professional APCs</cell_type> ( e.g . <cell_type>fibroblasts</cell_type> ) .


We have recently characterised a new factor called IK which is an efficient inhibitor of IFN-gamma induction of MHC class II antigens expression .

We have recently characterised a new factor called <protein>IK</protein> which is an efficient inhibitor of <protein>IFN-gamma</protein> induction of <protein>MHC class II antigens</protein> expression .


Here , we demonstrate a novel role for IK in MHC class II expression since over-expression of this protein by stable transfection into human B cells led to a total disappearance of constitutive MHC class II mRNA expression .

Here , we demonstrate a novel role for <protein>IK</protein> in <protein>MHC class II</protein> expression since over-expression of this protein by stable transfection into human B cells led to a total disappearance of constitutive <rna>MHC class II mRNA</rna> expression .


The class II transactivator ( CIITA ) is necessary for both constitutive and IFN-gamma induced MHC class II expressions .

The <dna>class II transactivator</dna> ( <dna>CIITA</dna> ) is necessary for both constitutive and <protein>IFN-gamma</protein> induced <protein>MHC class II</protein> expressions .


Examination of CIITA mRNA in IK stably transfected clones revealed a marked reduction of CIITA mRNA transcription .

Examination of <rna>CIITA mRNA</rna> in <protein>IK</protein> stably transfected clones revealed a marked reduction of <rna>CIITA mRNA</rna> transcription .


Taken together these results demonstrate that the IK protein plays a key role in the constitutive expression of MHC class II antigens and that inhibition induced by IK is upstream of CIITA in this regulatory pathway .

Taken together these results demonstrate that the <protein>IK protein</protein> plays a key role in the constitutive expression of <protein>MHC class II antigens</protein> and that inhibition induced by <protein>IK</protein> is upstream of <dna>CIITA</dna> in this regulatory pathway .


Selective expression of an interleukin-12 receptor component by human T helper 1 cells .

Selective expression of an <protein>interleukin-12 receptor component</protein> by <cell_type>human T helper 1 cells</cell_type> .


Interleukin-12 ( IL-12 ) , a heterodimeric cytokine produced by activated monocytes and dendritic cells , plays a crucial role in regulating interferon ( IFN ) -gamma production and in the generation of IFN-gamma-producing T helper 1 ( Th1 ) cells .

<protein>Interleukin-12</protein> ( <protein>IL-12</protein> ) , a <protein>heterodimeric cytokine</protein> produced by activated monocytes and dendritic cells , plays a crucial role in regulating <protein>interferon ( IFN ) -gamma</protein> production and in the generation of <cell_type>IFN-gamma-producing T helper 1 ( Th1 ) cells</cell_type> .


Here we show that the IL-12 receptor ( IL-12R ) beta 2 subunit , a recently cloned binding and signal transducing component of the IL-12R , is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway .

Here we show that the <protein>IL-12 receptor ( IL-12R ) beta 2 subunit</protein> , a recently cloned <protein>binding and signal transducing component</protein> of the <protein>IL-12R</protein> , is expressed on <cell_line>human Th1 but not Th2 clones</cell_line> and is induced during differentiation of <cell_type>human naive cells</cell_type> along the Th1 but not the Th2 pathway .


IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering .

<protein>IL-12</protein> and <protein>type I but not type II interferons</protein> induce expression of the <protein>IL-12R beta 2 chain</protein> during in vitro T cell differentiation after antigen receptor triggering .


The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies .

The selective expression and regulation of the <protein>IL-12R beta 2 subunit</protein> may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies .


The T cell activation factor NF-ATc positively regulates HIV-1 replication and gene expression in T cells .

The <protein>T cell activation factor</protein> <protein>NF-ATc</protein> positively regulates HIV-1 replication and gene expression in <cell_type>T cells</cell_type> .


Clinical deterioration in human immunodeficiency virus type 1 ( HIV-1 ) infection is associated with increased levels of viral replication and burden in the peripheral blood and lymphoid organs .


T cell activation and ensuing cellular gene activation can be critical for HIV-1 replication .


The hypothesis that the nuclear factor of activated T cells ( NF-AT ) may influence HIV-1 replication is therefore compelling given the tight correlation of HIV-1 transcriptional induction to T cell activation .

The hypothesis that the <protein>nuclear factor of activated T cells</protein> ( <protein>NF-AT</protein> ) may influence HIV-1 replication is therefore compelling given the tight correlation of HIV-1 transcriptional induction to T cell activation .


We report that certain NF-AT ( Rel ) family members productively bind the kappaB regulatory elements , synergize with NF-kappaB and Tat in transcriptional activation of HIV-1 , and enhance HIV-1 replication in T cells .

We report that certain <protein>NF-AT ( Rel ) family members</protein> productively bind the <dna>kappaB regulatory elements</dna> , synergize with <protein>NF-kappaB</protein> and <protein>Tat</protein> in transcriptional activation of HIV-1 , and enhance HIV-1 replication in <cell_type>T cells</cell_type> .


These results link regulatory factors critical to T cell commitment directly to HIV-1 replication .

These results link <protein>regulatory factors</protein> critical to T cell commitment directly to HIV-1 replication .


Transcription mediated by NFAT is highly inducible in effector CD4+ T helper 2 ( Th2 ) cells but not in Th1 cells .

Transcription mediated by <protein>NFAT</protein> is highly inducible in effector <cell_type>CD4+ T helper 2 ( Th2 ) cells</cell_type> but not in <cell_type>Th1 cells</cell_type> .


Transcriptional factors of the NFAT family play an important role in regulating the expression of several cytokine genes during the immune response , such as the genes for interleukin 2 ( IL-2 ) and IL-4 , among others .

<protein>Transcriptional factors</protein> of the <protein>NFAT family</protein> play an important role in regulating the expression of several <dna>cytokine genes</dna> during the immune response , such as the genes for <protein>interleukin 2</protein> ( <protein>IL-2</protein> ) and <protein>IL-4</protein> , among others .


Upon antigen stimulation , precursor CD4+ T helper ( pTh ) cells proliferate and differentiate into two populations of effector cells ( eTh1 and eTh2 ) , each one expressing a specific pattern of cytokines that distinguishes them from their precursors .

Upon antigen stimulation , precursor <cell_type>CD4+ T helper ( pTh ) cells</cell_type> proliferate and differentiate into two populations of <cell_type>effector cells</cell_type> ( <cell_type>eTh1</cell_type> and <cell_type>eTh2</cell_type> ) , each one expressing a specific pattern of <protein>cytokines</protein> that distinguishes them from their precursors .


eTh2 cells are the major source of IL-4 , while gamma interferon is produced by eTh1 cells .

<cell_type>eTh2 cells</cell_type> are the major source of <protein>IL-4</protein> , while <protein>gamma interferon</protein> is produced by <cell_type>eTh1 cells</cell_type> .


Here we have used reporter transgenic mice to show that DNA binding and transcriptional activities of NFAT are transiently induced during the differentiation of pTh cells into either eTh1 or eTh2 cells to mediate the expression of IL-2 as a common growth factor in both pathways .

Here we have used reporter transgenic mice to show that DNA binding and transcriptional activities of <protein>NFAT</protein> are transiently induced during the differentiation of <cell_type>pTh cells</cell_type> into either <cell_type>eTh1</cell_type> or <cell_type>eTh2 cells</cell_type> to mediate the expression of <protein>IL-2</protein> as a common growth factor in both pathways .


However , although NFAT DNA binding is similarly induced in both eTh1 and eTh2 cells upon antigen stimulation , only the NFAT complexes present in eTh2 cells are able to mediate high-level transcription , and relatively little NFAT transcriptional activity was induced in eTh1 cells .

However , although <protein>NFAT</protein> DNA binding is similarly induced in both <cell_type>eTh1</cell_type> and <cell_type>eTh2 cells</cell_type> upon antigen stimulation , only the <protein>NFAT complexes</protein> present in <cell_type>eTh2 cells</cell_type> are able to mediate high-level transcription , and relatively little <protein>NFAT</protein> transcriptional activity was induced in <cell_type>eTh1 cells</cell_type> .


In contrast to activated pTh cells , neither eTh1 nor eTh2 cells produced significant IL-2 upon stimulation , but the high levels of NFAT transcriptional activities directly correlate with the IL-4 production induced in response to antigen stimulation in eTh2 cells .

In contrast to activated <cell_type>pTh cells</cell_type> , neither <cell_type>eTh1</cell_type> nor <cell_type>eTh2 cells</cell_type> produced significant <protein>IL-2</protein> upon stimulation , but the high levels of <protein>NFAT</protein> transcriptional activities directly correlate with the <protein>IL-4</protein> production induced in response to antigen stimulation in <cell_type>eTh2 cells</cell_type> .


These data suggest that activated NFAT is involved in the effector function of eTh2 cells and that the failure of eTh1 cells to produce IL-4 in response to an antigen is due , at least partially , to a failure to induce high-level transcription of the IL-4 gene by NFAT .

These data suggest that activated <protein>NFAT</protein> is involved in the effector function of <cell_type>eTh2 cells</cell_type> and that the failure of <cell_type>eTh1 cells</cell_type> to produce <protein>IL-4</protein> in response to an antigen is due , at least partially , to a failure to induce high-level transcription of the <dna>IL-4 gene</dna> by <protein>NFAT</protein> .


Regulation of NFAT could be therefore a critical element in the polarization to eTh1 or eTh2 .

Regulation of <protein>NFAT</protein> could be therefore a critical element in the polarization to <cell_type>eTh1</cell_type> or <cell_type>eTh2</cell_type> .


Activation of the transcription factor NF-kappaB in lipopolysaccharide-stimulated U937 cells .

Activation of the <protein>transcription factor</protein> <protein>NF-kappaB</protein> in <cell_line>lipopolysaccharide-stimulated U937 cells</cell_line> .


During the course of serious bacterial infections , lipopolysaccharide ( LPS ) interacts with monocyte/macrophage receptors , resulting in the generation of inflammatory cytokines .

During the course of serious bacterial infections , lipopolysaccharide ( LPS ) interacts with <protein>monocyte/macrophage receptors</protein> , resulting in the generation of <protein>inflammatory cytokines</protein> .


Transcription factor NF-kappaB is crucial in activating the transcription of genes encoding proinflammatory cytokines .

<protein>Transcription factor</protein> <protein>NF-kappaB</protein> is crucial in activating the transcription of genes encoding <protein>proinflammatory cytokines</protein> .


In this paper , we demonstrate that the activation of NF-kappaB by LPS in a promonocytic cell line ( U937 ) followed a rather slow kinetics , depending on the rate of IkappaB-alpha inhibitor hydrolysis .

In this paper , we demonstrate that the activation of <protein>NF-kappaB</protein> by LPS in a <cell_line>promonocytic cell line</cell_line> ( <cell_line>U937</cell_line> ) followed a rather slow kinetics , depending on the rate of <protein>IkappaB-alpha</protein> inhibitor hydrolysis .


No degradation of p105 and p100 inhibitors was observed under these conditions .

No degradation of <protein>p105 and p100 inhibitors</protein> was observed under these conditions .


The transduction pathway leading to NF-kappaB activation in U937 cells involved the intracellular generation of reactive oxygen species ( ROS ) , as demonstrated by the concomitant inhibitory effects of antioxidants on NF-kappaB activation and the emission of a fluorescent probe reacting intracellularly with hydrogen peroxide .

The transduction pathway leading to <protein>NF-kappaB</protein> activation in <cell_line>U937 cells</cell_line> involved the intracellular generation of reactive oxygen species ( ROS ) , as demonstrated by the concomitant inhibitory effects of antioxidants on <protein>NF-kappaB</protein> activation and the emission of a fluorescent probe reacting intracellularly with hydrogen peroxide .


This ROS pathway was also characterized by the use of other inhibitors .


This finding indicates that phospholipase A2 and 5-lipoxygenase are also involved .

This finding indicates that <protein>phospholipase A2</protein> and <protein>5-lipoxygenase</protein> are also involved .


However , the NF-kappaB activation pathway involving the acidic sphingomyelinase of the endolysosomial membrane did not seem to participate in the LPS-induced NF-kappaB activation in U937 cells .

However , the <protein>NF-kappaB</protein> activation pathway involving the <protein>acidic sphingomyelinase</protein> of the endolysosomial membrane did not seem to participate in the LPS-induced <protein>NF-kappaB</protein> activation in <cell_line>U937 cells</cell_line> .


c-Rel is a target of pentoxifylline-mediated inhibition of T lymphocyte activation .

<protein>c-Rel</protein> is a target of pentoxifylline-mediated inhibition of T lymphocyte activation .


The possible clinical use of the methyl xanthine derivative , pentoxifylline ( PF ) , for the treatment of T cell-dependent diseases is being noted with increasing interest .


In this paper , we studied the molecular consequences of PF treatment during lymphocyte activation .


We found that in T cells , anti-CD3-induced c-Rel expression was blocked by PF , whereas the induction of other NF-kappaB family members was not significantly affected .

We found that in <cell_type>T cells</cell_type> , anti-CD3-induced <protein>c-Rel</protein> expression was blocked by PF , whereas the induction of other <protein>NF-kappaB family members</protein> was not significantly affected .


However , induction of NF-AT , which has the same signaling requirements as c-Rel induction , was not inhibited by PF .

However , induction of <protein>NF-AT</protein> , which has the same signaling requirements as <protein>c-Rel</protein> induction , was not inhibited by PF .


Among genes that respond to these transcription factors , IL-2 mRNA induction was suppressed by PF , whereas IL-2R ( alpha ) chain mRNA induction was not affected .

Among genes that respond to these <protein>transcription factors</protein> , <rna>IL-2 mRNA</rna> induction was suppressed by PF , whereas <rna>IL-2R ( alpha ) chain mRNA</rna> induction was not affected .


These observations implicated c-Rel as an IL-2 promoter factor , for which experimental support was obtained from transient transfection experiments .

These observations implicated <protein>c-Rel</protein> as an <protein>IL-2 promoter factor</protein> , for which experimental support was obtained from transient transfection experiments .


In contrast with the observation in T cells , c-Rel induction was not blocked by PF in B cells .

In contrast with the observation in <cell_type>T cells</cell_type> , <protein>c-Rel</protein> induction was not blocked by PF in <cell_type>B cells</cell_type> .


The greater selectivity of PF , compared with FK506 , at both the molecular and cellular levels may prove advantageous in manipulating T cell responses in vivo .


V3 loop of human immunodeficiency virus type 1 suppresses interleukin 2 -induced T cell growth [ published erratum appears in AIDS Res Hum Retroviruses 1997 May 1 ; 13 ( 7 ) : 633 ]

<protein>V3 loop</protein> of human immunodeficiency virus type 1 suppresses <protein>interleukin 2</protein> -induced T cell growth [ published erratum appears in AIDS Res Hum Retroviruses 1997 May 1 ; 13 ( 7 ) : 633 ]


We tested the effect of three linear or two loop peptides derived from the V3 region of the HTLV-III BH10 clone or the SF2 strain of human immunodeficiency virus type 1 on IL-2 -driven T cell proliferation .

We tested the effect of three linear or two loop peptides derived from the <protein>V3 region</protein> of the HTLV-III BH10 clone or the SF2 strain of human immunodeficiency virus type 1 on <protein>IL-2</protein> -driven T cell proliferation .


V3-BH10 , which consists of 42 amino acids and has a loop structure , suppressed IL-2 -driven proliferation of all IL-2-dependent cells [ Kit225 , ED-40515 ( + ) , KT-3 , 7-day PHA-blasts , and fresh peripheral blood mononuclear cells ] tested , whereas it did not suppress the cell growth of IL-2-independent cell lines ( Hut102 , Molt-4 , and Jurkat ) .

<protein>V3-BH10</protein> , which consists of <protein>42 amino acids</protein> and has a <protein>loop structure</protein> , suppressed <protein>IL-2</protein> -driven proliferation of all <cell_line>IL-2-dependent cells</cell_line> [ <cell_line>Kit225</cell_line> , <cell_line>ED-40515 ( + )</cell_line> , <cell_line>KT-3</cell_line> , <cell_line>7-day PHA-blasts</cell_line> , and <cell_type>fresh peripheral blood mononuclear cells</cell_type> ] tested , whereas it did not suppress the cell growth of <cell_line>IL-2-independent cell lines</cell_line> ( <cell_line>Hut102</cell_line> , <cell_line>Molt-4</cell_line> , and <cell_line>Jurkat</cell_line> ) .


This suppressive effect was also seen in IL-2 -driven cell growth of CD8-positive lymphocytes purified from 7-day PHA-blasts , indicating that CD4 molecules were not required for the suppression .

This suppressive effect was also seen in <protein>IL-2</protein> -driven cell growth of <cell_type>CD8-positive lymphocytes</cell_type> purified from <cell_type>7-day PHA-blasts</cell_type> , indicating that <protein>CD4 molecules</protein> were not required for the suppression .


The treatment with anti-V3 loop monoclonal antibody ( 902 antibody ) completely abolished the suppressive effect of V3-BH10 .

The treatment with <protein>anti-V3 loop monoclonal antibody</protein> ( <protein>902 antibody</protein> ) completely abolished the suppressive effect of <protein>V3-BH10</protein> .


In addition , V3-BH10 generated the arrest of Kit225 cells and also purified CD8-positive lymphocytes in G1 phase in the presence of IL-2 .

In addition , <protein>V3-BH10</protein> generated the arrest of <cell_line>Kit225 cells</cell_line> and also <cell_line>purified CD8-positive lymphocytes</cell_line> in G1 phase in the presence of <protein>IL-2</protein> .


Neither chromatin condensation nor DNA fragmentation was detected in Kit225 cells cultured with V3-BH10 and IL-2 .

Neither <dna>chromatin</dna> condensation nor DNA fragmentation was detected in <cell_line>Kit225 cells</cell_line> cultured with <protein>V3-BH10</protein> and <protein>IL-2</protein> .


V3-BH10 neither blocked radiolabeled IL-2 binding to IL-2 receptors nor affected tyrosyl phosphorylation of several cellular proteins ( p120 , p98 , p96 , p54 , and p38 ) , which is immediately induced by IL-2 stimulation .

<protein>V3-BH10</protein> neither blocked radiolabeled <protein>IL-2</protein> binding to <protein>IL-2 receptors</protein> nor affected tyrosyl phosphorylation of several <protein>cellular proteins</protein> ( <protein>p120</protein> , <protein>p98</protein> , <protein>p96</protein> , <protein>p54</protein> , and <protein>p38</protein> ) , which is immediately induced by <protein>IL-2</protein> stimulation .


However , V3-BH10 enhanced IL-2 -induced mRNA expression of c-fos but not c-myc or junB .

However , <protein>V3-BH10</protein> enhanced <protein>IL-2</protein> <rna>-induced mRNA</rna> expression of <dna>c-fos</dna> but not <dna>c-myc</dna> or <dna>junB</dna> .


Thus , the binding of V3 loop of gp120 to the cell surface molecule ( s ) appears to affect intracellular IL-2 signaling , which leads to the suppression of IL-2 -induced T cell growth

Thus , the binding of <protein>V3 loop</protein> of <protein>gp120</protein> to the cell surface molecule ( s ) appears to affect intracellular <protein>IL-2</protein> signaling , which leads to the suppression of <protein>IL-2</protein> -induced T cell growth


A negative role for phosphoinositide 3-kinase in T-cell antigen receptor function .

A negative role for <protein>phosphoinositide 3-kinase</protein> in <protein>T-cell antigen receptor</protein> function .


BACKGROUND : A delicate balance between positive and negative regulatory mechanisms during T-cell activation determines the specificity and magnitude of an immune response .


Phosphoinositide 3-kinase ( PI 3-kinase ) is activated by a diverse set of receptors that determine T-cell function , including the T-cell antigen receptor ( TCR ) , the costimulatory receptor CD28 , and negative regulators of T-cell activation such as CTLA-4 .

<protein>Phosphoinositide 3-kinase</protein> ( <protein>PI 3-kinase</protein> ) is activated by a diverse set of receptors that determine T-cell function , including the <protein>T-cell antigen receptor</protein> ( <protein>TCR</protein> ) , the <protein>costimulatory receptor CD28</protein> , and <protein>negative regulators</protein> of T-cell activation such as <protein>CTLA-4</protein> .


PI 3-kinase is also regulated by the haematopoietic cytokines that determine T-cell differentiation and lymphocyte proliferation .

<protein>PI 3-kinase</protein> is also regulated by the haematopoietic cytokines that determine T-cell differentiation and <cell_type>lymphocyte</cell_type> proliferation .


PI 3-kinase can thus dynamically influence the outcome of the immune reactions at various stages .

<protein>PI 3-kinase</protein> can thus dynamically influence the outcome of the immune reactions at various stages .


In this study , we investigated the importance of PI 3-kinase in TCR -directed T-cell activation using activated or inhibitory versions of PI 3-kinase .

In this study , we investigated the importance of <protein>PI 3-kinase</protein> in <protein>TCR</protein> -directed T-cell activation using activated or inhibitory versions of <protein>PI 3-kinase</protein> .


RESULTS : Certain aspects of TCR responses such as the induction of transcriptional activity of AP1 and serum response factor were not affected by expression of the mutant forms of PI 3-kinase .

RESULTS : Certain aspects of <protein>TCR</protein> responses such as the induction of transcriptional activity of <protein>AP1</protein> and <protein>serum response factor</protein> were not affected by expression of the mutant forms of <protein>PI 3-kinase</protein> .


We found , however , that PI 3-kinase profoundly influenced the transactivation capacity of ' nuclear factor of activated T cells ' ( NF-AT ) elicited by the TCR : expression of an activated form of PI 3-kinase inhibited TCR -mediated NF-AT responses , whereas expression of a dominant negative mutant of PI 3-kinase potently enhanced TCR -controlled NF-AT induction .

We found , however , that <protein>PI 3-kinase</protein> profoundly influenced the transactivation capacity of ' <protein>nuclear factor of activated T cells</protein> ' ( <protein>NF-AT</protein> ) elicited by the <protein>TCR</protein> : expression of an activated form of <protein>PI 3-kinase</protein> inhibited <protein>TCR</protein> -mediated <protein>NF-AT</protein> responses , whereas expression of a dominant negative mutant of <protein>PI 3-kinase</protein> potently enhanced <protein>TCR</protein> -controlled <protein>NF-AT</protein> induction .


These effects of PI 3-kinase were not mediated by previously identified PI 3-kinase effectors , such as protein kinase B , a positive regulator of PI 3-kinase , or the GTPase Rac , and are therefore likely to involve a novel , as yet unknown , effector molecule .

These effects of <protein>PI 3-kinase</protein> were not mediated by previously identified <protein>PI 3-kinase effectors</protein> , such as <protein>protein kinase B</protein> , a <protein>positive regulator</protein> of <protein>PI 3-kinase</protein> , or the <protein>GTPase Rac</protein> , and are therefore likely to involve a novel , as yet unknown , <protein>effector molecule</protein> .


CONCLUSIONS : Our results establish that PI 3-kinase can both positively and negatively regulate T-cell function , and uncover a previously unrecognized function for PI 3-kinase in T cells as a selective negative regulator of TCR -signalling events and therefore as a determinant of T-cell homeostasis .

CONCLUSIONS : Our results establish that <protein>PI 3-kinase</protein> can both positively and negatively regulate T-cell function , and uncover a previously unrecognized function for <protein>PI 3-kinase</protein> in <cell_type>T cells</cell_type> as a selective negative regulator of <protein>TCR</protein> -signalling events and therefore as a determinant of T-cell homeostasis .


The Pax-5 gene is alternatively spliced during B-cell development .

The <dna>Pax-5 gene</dna> is alternatively spliced during B-cell development .


The transcription factor Pax-5 is expressed during the early stages of B-cell differentiation and influences the expression of several B-cell-specific genes .

The <protein>transcription factor Pax-5</protein> is expressed during the early stages of B-cell differentiation and influences the expression of several <dna>B-cell-specific genes</dna> .


In addition to the existing isoform ( Pax-5 , which we have named Pax-5a ) , we have isolated three new isoforms , Pax-5b , Pax-5d , and Pax-5e , from murine spleen and B-lymphoid cell lines using library screenings and polymerase chain reaction amplification .

In addition to the existing isoform ( <protein>Pax-5</protein> , which we have named <protein>Pax-5a</protein> ) , we have isolated three new isoforms , <protein>Pax-5b</protein> , <protein>Pax-5d</protein> , and <protein>Pax-5e</protein> , from murine spleen and <cell_line>B-lymphoid cell lines</cell_line> using library screenings and polymerase chain reaction amplification .


Isoforms Pax-5b and Pax-5e have spliced out their second exon , resulting in proteins with only a partial DNA-binding domain .

Isoforms <protein>Pax-5b</protein> and <protein>Pax-5e</protein> have spliced out their second exon , resulting in proteins with only a partial <protein>DNA-binding domain</protein> .


Isoforms Pax-5d and Pax-5e have deleted the 3'-region , which encodes the transactivating domain , and replaced it with a novel sequence .

Isoforms Pax-5d and <protein>Pax-5e</protein> have deleted the <protein>3'-region</protein> , which encodes the transactivating domain , and replaced it with a novel sequence .


The existence of alternative Pax-5 transcripts was confirmed using RNase protection assays .

The existence of alternative <rna>Pax-5 transcripts</rna> was confirmed using <protein>RNase</protein> protection assays .


Furthermore , Pax-5a and Pax-5b proteins were detected using Western blot analysis .

Furthermore , <protein>Pax-5a</protein> and <protein>Pax-5b proteins</protein> were detected using Western blot analysis .


Pax-5a was detectable in pro- , pre- , and mature B-cell lines , but not in two plasmacytomas ; Pax-5b was shown to be present at low levels in mature B-cell lines and , unexpectedly , in one plasma cell line , but not in pro-B-cell or T-cell lines .

<protein>Pax-5a</protein> was detectable in <cell_line>pro- , pre- , and mature B-cell lines</cell_line> , but not in two plasmacytomas ; <protein>Pax-5b</protein> was shown to be present at low levels in <cell_line>mature B-cell lines</cell_line> and , unexpectedly , in one plasma cell line , but not in <cell_line>pro-B-cell or T-cell lines</cell_line> .


Mobility shift assays showed that in vitro translated Pax-5a and Pax-5d , but not Pax-5b or Pax-5e , could interact with a B-cell-specific activator protein-binding site on the blk promoter .

Mobility shift assays showed that in vitro translated <protein>Pax-5a</protein> and <protein>Pax-5d</protein> , but not <protein>Pax-5b</protein> or <protein>Pax-5e</protein> , could interact with a <dna>B-cell-specific activator protein-binding site</dna> on the <dna>blk promoter</dna> .


Using this assay , we also showed that Pax-5d was present in nuclear extracts of some ( but not all ) B-lymphoid lines and interacts with the B-cell-specific activator protein-binding site .

Using this assay , we also showed that <protein>Pax-5d</protein> was present in nuclear extracts of some ( but not all ) <cell_line>B-lymphoid lines</cell_line> and interacts with the <dna>B-cell-specific activator protein-binding site</dna> .


The pattern of differential expression of alternatively spliced Pax-5 isoforms suggests that they may be important regulators of transcription during B-cell maturation .

The pattern of differential expression of <protein>alternatively spliced Pax-5 isoforms</protein> suggests that they may be important <protein>regulators of transcription</protein> during B-cell maturation .


Reconstitution of T cell antigen receptor-induced Erk2 kinase activation in Lck-negative JCaM1 cells by Syk .

Reconstitution of T cell antigen receptor-induced <protein>Erk2 kinase</protein> activation in <cell_line>Lck-negative JCaM1 cells</cell_line> by <protein>Syk</protein> .


The two related protein-tyrosine kinases Syk and Zap are rapidly phosphorylated on tyrosine residues and enzymatically activated upon crosslinking of the T cell antigen receptor .

The two related <protein>protein-tyrosine kinases</protein> <protein>Syk</protein> and <protein>Zap</protein> are rapidly phosphorylated on tyrosine residues and enzymatically activated upon crosslinking of the <protein>T cell antigen receptor</protein> .


We have previously reported that the activation of Syk is less dependent on the Src family kinase Lck than the activation of Zap .

We have previously reported that the activation of <protein>Syk</protein> is less dependent on the <protein>Src family kinase</protein> <protein>Lck</protein> than the activation of <protein>Zap</protein> .


Here we report that overexpression of Syk in the Lck-negative JCaM1 cells enabled the T cell antigen receptor/CD3 complex to induce a normal activation of the mitogen-activated protein kinase ( MAPK ) pathway and expression of a nuclear factor of activated T cells reporter construct .

Here we report that overexpression of <protein>Syk</protein> in the <cell_line>Lck-negative JCaM1 cells</cell_line> enabled the <protein>T cell antigen receptor/CD3 complex</protein> to induce a normal activation of the <protein>mitogen-activated protein kinase</protein> ( <protein>MAPK</protein> ) pathway and expression of a <protein>nuclear factor of activated T cells</protein> reporter construct .


In contrast , Zap and other protein-tyrosine kinases were unable to reconstitute these signaling pathways when expressed at the same levels .

In contrast , <protein>Zap</protein> and other <protein>protein-tyrosine kinases</protein> were unable to reconstitute these signaling pathways when expressed at the same levels .


In parallel , Syk was phosphorylated on tyrosine , while Zap was not .

In parallel , <protein>Syk</protein> was phosphorylated on tyrosine , while <protein>Zap</protein> was not .


The Syk -mediated T cell antigen receptor -induced MAPK activation was detectable within 1 min of receptor stimulation and peaked at 3-5 min .

The <protein>Syk</protein> -mediated <protein>T cell antigen receptor</protein> -induced <protein>MAPK</protein> activation was detectable within 1 min of receptor stimulation and peaked at 3-5 min .


The capacity of Syk to reconstitute the MAPK response required the catalytic activity of Syk , an intact autophosphorylation site ( Y518 and Y519 ) , both Src homology 2 domains and it was blocked by the inhibitory N17-mutated dominant-negative Ras construct .

The capacity of <protein>Syk</protein> to reconstitute the <protein>MAPK</protein> response required the catalytic activity of <protein>Syk</protein> , an intact <protein>autophosphorylation site</protein> ( Y518 and Y519 ) , both <protein>Src homology 2 domains</protein> and it was blocked by the <protein>inhibitory N17-mutated dominant-negative Ras construct</protein> .


A Y341 -- > F mutant of Syk , which is deficient in its interaction with phospholipase Cy1 and Vav , was less efficient than wild-type Syk .

A Y341 -- > <protein>F mutant</protein> of <protein>Syk</protein> , which is deficient in its interaction with <protein>phospholipase Cy1</protein> and <protein>Vav</protein> , was less efficient than <protein>wild-type Syk</protein> .


Our results suggest that Syk , in contrast to Zap , can transduce signals from the T cell antigen receptor independently of Lck .

Our results suggest that <protein>Syk</protein> , in contrast to <protein>Zap</protein> , can transduce signals from the <protein>T cell antigen receptor</protein> independently of <protein>Lck</protein> .


Activation of the transcription factor MEF2C by the MAP kinase p38 in inflammation .

Activation of the <protein>transcription factor</protein> <protein>MEF2C</protein> by the <protein>MAP kinase p38</protein> in inflammation .


For cells of the innate immune system to mount a host defence response to infection , they must recognize products of microbial pathogens such as lipopolysaccharide ( LPS ) , the endotoxin secreted by Gram-negative bacteria .


These cellular responses require intracellular signalling pathways , such as the four MAP kinase ( MAPK ) pathways .

These cellular responses require intracellular signalling pathways , such as the four <protein>MAP kinase</protein> ( <protein>MAPK</protein> ) pathways .


In mammalian cells the MAPK p38 is thought to play an important role in the regulation of cellular responses during infection through its effects on the expression of proinflammatory molecules .

In mammalian cells the <protein>MAPK p38</protein> is thought to play an important role in the regulation of cellular responses during infection through its effects on the expression of <protein>proinflammatory molecules</protein> .


One means of understanding the role of p38 in these responses is to identify proteins with functions regulated by p38 -catalysed phosphorylation .

One means of understanding the role of <protein>p38</protein> in these responses is to identify proteins with functions regulated by <protein>p38</protein> -catalysed phosphorylation .


Here we demonstrate a link between the p38 pathway and a member of the myocyte-enhancer factor 2 ( MEF2 ) group of transcription factors .

Here we demonstrate a link between the <protein>p38</protein> pathway and a member of the <protein>myocyte-enhancer factor 2 ( MEF2 ) group</protein> of <protein>transcription factors</protein> .


We found that in monocytic cells , LPS increases the transactivation activity of MEF2C through p38 -catalysed phosphorylation .

We found that in <cell_type>monocytic cells</cell_type> , LPS increases the transactivation activity of <protein>MEF2C</protein> through <protein>p38</protein> -catalysed phosphorylation .


One consequence of MEF2C activation is increased c-jun gene transcription .

One consequence of <protein>MEF2C</protein> activation is increased <dna>c-jun gene</dna> transcription .


Our results show that p38 may influence host defence and inflammation by maintaining the balance of c-Jun protein consumed during infection .

Our results show that <protein>p38</protein> may influence host defence and inflammation by maintaining the balance of <protein>c-Jun protein</protein> consumed during infection .


Cellular redox status influences both cytotoxic and NF-kappa B activation in natural killer cells .

Cellular redox status influences both cytotoxic and NF-kappa B activation in <cell_type>natural killer cells</cell_type> .


The role of cellular redox status in both cytotoxic activity and NF-kappa B activation in natural killer ( NK ) cells was investigated .

The role of cellular redox status in both cytotoxic activity and <protein>NF-kappa B</protein> activation in <cell_type>natural killer ( NK ) cells</cell_type> was investigated .


The results indicate that stimulation of NK cells , either freshly isolated from peripheral blood lymphocytes ( PBL ) or long-term cultured NK clones , with specific cell targets results in an increased binding activity of NF-kappa B and AP-1 transcription factors measured by gel retardation .

The results indicate that stimulation of <cell_type>NK cells</cell_type> , either freshly isolated from <cell_type>peripheral blood lymphocytes</cell_type> ( <cell_type>PBL</cell_type> ) or <cell_line>long-term cultured NK clones</cell_line> , with specific cell targets results in an increased binding activity of <protein>NF-kappa B</protein> and <protein>AP-1</protein> <protein>transcription factors</protein> measured by gel retardation .


Pretreatment of NK cells with the antioxidant pyrrolidine dithiocarbarmate ( PDTC ) leads to the inhibition of NF-kappa B activation but the AP-1 binding to DNA was superinduced .

Pretreatment of <cell_type>NK cells</cell_type> with the antioxidant pyrrolidine dithiocarbarmate ( PDTC ) leads to the inhibition of <protein>NF-kappa B</protein> activation but the <protein>AP-1</protein> binding to DNA was superinduced .


The inhibition of NF-kappa B by PDTC paralleled with an inhibition of spontaneous cytotoxicity mediated by NK cells .

The inhibition of <protein>NF-kappa B</protein> by PDTC paralleled with an inhibition of spontaneous cytotoxicity mediated by <cell_type>NK cells</cell_type> .


Moreover , the inhibitors of serine proteases , N-alpha-tosyl-L-lysine chloromethyl ketone and N-alpha-tosyl-L-phenylalanine chloromethyl ketone , also blocked the cytolytic activity of NK cells against the sensitive target K562 .

Moreover , the inhibitors of <protein>serine proteases</protein> , N-alpha-tosyl-L-lysine chloromethyl ketone and N-alpha-tosyl-L-phenylalanine chloromethyl ketone , also blocked the cytolytic activity of <cell_type>NK cells</cell_type> against the sensitive target <cell_line>K562</cell_line> .


In contrast , NK activity was not affected by pretreatment of the effector cells with the proteasome inhibitor N-acetyl-leu-leu-norleucinal which selectively inhibits NF-kappa B activation .

In contrast , NK activity was not affected by pretreatment of the effector cells with the <protein>proteasome</protein> inhibitor N-acetyl-leu-leu-norleucinal which selectively inhibits <protein>NF-kappa B</protein> activation .


Altogether , these results support the hypothesis that the activation of NK cells involved transcriptional and post-transcriptional events , and that reactive intermediates may play an important role in the molecular processes related with the generation of a cytotoxic response by NK cells .

Altogether , these results support the hypothesis that the activation of <cell_type>NK cells</cell_type> involved transcriptional and post-transcriptional events , and that reactive intermediates may play an important role in the molecular processes related with the generation of a cytotoxic response by <cell_type>NK cells</cell_type> .


The predominant E2F complex in human primary haemopoietic cells and in AML blasts contains E2F-4 , DP-1 and p130 .

The predominant <protein>E2F complex</protein> in <cell_type>human primary haemopoietic cells</cell_type> and in <cell_type>AML blasts</cell_type> contains <protein>E2F-4</protein> , <protein>DP-1</protein> and <protein>p130</protein> .


The E2F family of transcription factors are thought to play an important role in the control of cell cycle progression .

The <protein>E2F family</protein> of <protein>transcription factors</protein> are thought to play an important role in the control of cell cycle progression .


There is now also increasing evidence that some family members may act as oncogenes or tumour suppressor genes .

There is now also increasing evidence that some family members may act as <dna>oncogenes</dna> or <dna>tumour suppressor genes</dna> .


The characterization of these proteins in human primary haemopoietic cells and acute myeloid leukaemia ( AML ) blasts may thus give an insight to the molecular mechanisms governing proliferation and leukaemogenesis in these cells .

The characterization of these proteins in <cell_type>human primary haemopoietic cells</cell_type> and <cell_type>acute myeloid leukaemia ( AML ) blasts</cell_type> may thus give an insight to the molecular mechanisms governing proliferation and leukaemogenesis in these cells .


Therefore we analysed the expression of E2F -DNA binding activity and the constituent proteins found in the complexes in human primary haemopoietic cells of various lineages .

Therefore we analysed the expression of <protein>E2F</protein> -DNA binding activity and the constituent proteins found in the complexes in <cell_type>human primary haemopoietic cells</cell_type> of various lineages .


We also studied blasts from 18 patients with acute myeloid leukaemia ( AML ) .


On electromobility shift assays ( EMSA ) a single E2F-DNA binding complex was detected in T cells , B cells and monocytes which was shown to contain E2F-4 , DP-1 and p130 , indicating that all quiescent haemopoietic cells have the same complex .

On electromobility shift assays ( EMSA ) a single <protein>E2F-DNA binding complex</protein> was detected in <cell_type>T cells</cell_type> , <cell_type>B cells</cell_type> and <cell_type>monocytes</cell_type> which was shown to contain <protein>E2F-4</protein> , <protein>DP-1</protein> and <protein>p130</protein> , indicating that all <cell_type>quiescent haemopoietic cells</cell_type> have the same complex .


Examination of 18 AML samples by EMSA revealed the presence of E2F binding and no gross abnormalities were detected .

Examination of 18 AML samples by EMSA revealed the presence of <protein>E2F</protein> binding and no gross abnormalities were detected .


An E2F-4/p130 complex was detected in representative samples of all FAB types analysed .

An <protein>E2F-4/p130 complex</protein> was detected in representative samples of all <protein>FAB</protein> types analysed .


Thus abnormalities of E2F function are unlikely to play a primary pathogenic role in AML .

Thus abnormalities of <protein>E2F</protein> function are unlikely to play a primary pathogenic role in AML .


Characterization of a mutant cell line that does not activate NF-kappaB in response to multiple stimuli .

Characterization of a <cell_line>mutant cell line</cell_line> that does not activate <protein>NF-kappaB</protein> in response to multiple stimuli .


Numerous genes required during the immune or inflammation response as well as the adhesion process are regulated by nuclear factor kappaB ( NF-kappaB ) .

Numerous genes required during the immune or inflammation response as well as the adhesion process are regulated by <protein>nuclear factor kappaB</protein> ( <protein>NF-kappaB</protein> ) .


Associated with its inhibitor , I kappaB , NF-kappaB resides as an inactive form in the cytoplasm .

Associated with its inhibitor , <protein>I kappaB</protein> , <protein>NF-kappaB</protein> resides as an inactive form in the cytoplasm .


Upon stimulation by various agents , I kappaB is proteolyzed and NF-kappaB translocates to the nucleus , where it activates its target genes .

Upon stimulation by various agents , <protein>I kappaB</protein> is proteolyzed and <protein>NF-kappaB</protein> translocates to the nucleus , where it activates its target genes .


The transduction pathways that lead to I kappaB inactivation remain poorly understood .

The transduction pathways that lead to <protein>I kappaB</protein> inactivation remain poorly understood .


In this study , we have characterized a cellular mutant , the 70/Z3-derived 1.3E2 murine pre-B cell line , that does not activate NF-kappaB in response to several stimuli .

In this study , we have characterized a cellular mutant , the <cell_line>70/Z3-derived 1.3E2 murine pre-B cell line</cell_line> , that does not activate <protein>NF-kappaB</protein> in response to several stimuli .


We demonstrate that upon stimulation by lipopolysaccharide , Taxol , phorbol myristate acetate , interleukin-1 , or double-stranded RNA , I kappaB alpha is not degraded , as a result of an absence of induced phosphorylation on serines 32 and 36 .

We demonstrate that upon stimulation by lipopolysaccharide , Taxol , phorbol myristate acetate , <protein>interleukin-1</protein> , or <rna>double-stranded RNA</rna> , <protein>I kappaB alpha</protein> is not degraded , as a result of an absence of induced phosphorylation on serines 32 and 36 .


Neither a mutation in I kappaB alpha nor a mutation in p50 or relA , the two major subunits of NF-kappaB in this cell line , accounts for this phosphorylation defect .

Neither a mutation in <protein>I kappaB alpha</protein> nor a mutation in <protein>p50</protein> or <protein>relA</protein> , the two major subunits of <protein>NF-kappaB</protein> in this cell line , accounts for this phosphorylation defect .


As well as culminating in the inducible phosphorylation of I kappaB alpha on serines 32 and 36 , all the stimuli that are inactive on 1.3E2 cells exhibit a sensitivity to the antioxidant pyrrolidine dithiocarbamate ( PDTC ) .

As well as culminating in the inducible phosphorylation of <protein>I kappaB</protein> alpha on serines 32 and 36 , all the stimuli that are inactive on <cell_line>1.3E2 cells</cell_line> exhibit a sensitivity to the antioxidant pyrrolidine dithiocarbamate ( PDTC ) .


In contrast , stimuli such as hyperosmotic shock or phosphatase inhibitors , which use PDTC-insensitive pathways , induce I kappaB alpha degradation in 1.3E2 .

In contrast , stimuli such as hyperosmotic shock or phosphatase inhibitors , which use PDTC-insensitive pathways , induce <protein>I kappaB</protein> alpha degradation in <cell_line>1.3E2</cell_line> .


Analysis of the redox status of 1.3E2 does not reveal any difference from wild-type 70Z/3 .

Analysis of the redox status of <cell_line>1.3E2</cell_line> does not reveal any difference from <cell_line>wild-type 70Z/3</cell_line> .


We also report that the human T-cell leukemia virus type 1 ( HTLV-1 ) -derived Tax trans-activator induces NF-kappaB activity in 1.3E2 , suggesting that this viral protein does not operate via the defective pathway .

We also report that the <protein>human T-cell leukemia virus type 1 ( HTLV-1 ) -derived Tax trans-activator</protein> induces <protein>NF-kappaB</protein> activity in <cell_line>1.3E2</cell_line> , suggesting that this viral protein does not operate via the defective pathway .


Finally , we show that two other I kappaB molecules , I kappaB beta and the recently identified I kappaB epsilon , are not degraded in the 1.3E2 cell line following stimulation .

Finally , we show that two other <protein>I kappaB molecules</protein> , <protein>I kappaB beta</protein> and the recently identified <protein>I kappaB epsilon</protein> , are not degraded in the <cell_line>1.3E2</cell_line> cell line following stimulation .


Our results demonstrate that 1.3E2 is a cellular transduction mutant exhibiting a defect in a step that is required by several different stimuli to activate NF-kappaB .

Our results demonstrate that <cell_line>1.3E2</cell_line> is a <protein>cellular transduction mutant</protein> exhibiting a defect in a step that is required by several different stimuli to activate <protein>NF-kappaB</protein> .


In addition , this analysis suggests a common step in the signaling pathways that trigger I kappaB alpha , I kappaB beta , and I kappaB epsilon degradation .

In addition , this analysis suggests a common step in the signaling pathways that trigger <protein>I kappaB alpha</protein> , <protein>I kappaB beta</protein> , and <protein>I kappaB epsilon</protein> degradation .


Involvement of Egr-1 /RelA synergy in distinguishing T cell activation from tumor necrosis factor-alpha -induced NF-kappa B1 transcription .

Involvement of <protein>Egr-1</protein> <protein>/RelA</protein> synergy in distinguishing T cell activation from <protein>tumor necrosis factor-alpha</protein> -induced <protein>NF-kappa B1</protein> transcription .


NF-kappa B is an important transcription factor required for T cell proliferation and other immunological functions .

<protein>NF-kappa B</protein> is an important <protein>transcription factor</protein> required for T cell proliferation and other immunological functions .


The NF-kappa B1 gene encodes a 105-kD protein that is the precursor of the p50 component of NF-kappa B .

The <dna>NF-kappa B1 gene</dna> encodes a <protein>105-kD protein</protein> that is the precursor of the <protein>p50 component</protein> of <protein>NF-kappa B</protein> .


Previously , we and others have demonstrated that NF-kappa B regulates the NF-kappa B1 gene .

Previously , we and others have demonstrated that <protein>NF-kappa B</protein> regulates the <dna>NF-kappa B1 gene</dna> .


In this manuscript we have investigated the molecular mechanisms by which T cell lines stimulated with phorbol 12-myristate 13-acetate ( PMA ) and phytohemagglutin ( PHA ) display significantly higher levels of NF-kappa B1 encoding transcripts than cells stimulated with tumor necrosis factor-alpha , despite the fact that both stimuli activate NF-kappa B .

In this manuscript we have investigated the molecular mechanisms by which <cell_line>T cell lines</cell_line> stimulated with phorbol 12-myristate 13-acetate ( PMA ) and <protein>phytohemagglutin</protein> ( <protein>PHA</protein> ) display significantly higher levels of <protein>NF-kappa B1</protein> encoding <rna>transcripts</rna> than cells stimulated with <protein>tumor necrosis factor-alpha</protein> , despite the fact that both stimuli activate <protein>NF-kappa B</protein> .


Characterization of the NF-kappa B1 promoter identified an Egr-1 site which was found to be essential for both the PMA/ PHA -mediated induction as well as the synergistic activation observed after the expression of the RelA subunit of NF-kappa B and Egr-1 .

Characterization of the <dna>NF-kappa B1 promoter</dna> identified an <dna>Egr-1 site</dna> which was found to be essential for both the PMA/ <protein>PHA</protein> -mediated induction as well as the synergistic activation observed after the expression of the RelA subunit of <protein>NF-kappa B</protein> and <protein>Egr-1</protein> .


Furthermore , Egr-1 induction was required for endogenous NF-kappa B1 gene expression , since PMA/ PHA-stimulated T cell lines expressing antisense Egr-1 RNA were inhibited in their ability to upregulate NF-kappa B1 transcription .

Furthermore , <protein>Egr-1</protein> induction was required for endogenous <dna>NF-kappa B1</dna> gene expression , since PMA/ <cell_line>PHA-stimulated T cell lines</cell_line> expressing antisense <rna>Egr-1 RNA</rna> were inhibited in their ability to upregulate NF-kappa B1 transcription .


Our studies indicate that transcriptional synergy mediated by activation of both Egr-1 and NF-kappa B may have important ramifications in T cell development by upregulating NF-kappa B1 gene expression .

Our studies indicate that transcriptional synergy mediated by activation of both <protein>Egr-1</protein> and <protein>NF-kappa B</protein> may have important ramifications in T cell development by upregulating <dna>NF-kappa B1</dna> gene expression .


The synthesis and immunogenicity of varicella-zoster virus glycoprotein E and immediate-early protein ( IE62 ) expressed in recombinant herpes simplex virus-1 .

The synthesis and immunogenicity of <protein>varicella-zoster virus glycoprotein E</protein> and <protein>immediate-early protein</protein> ( <protein>IE62</protein> ) expressed in recombinant herpes simplex virus-1 .


In order to evaluate the conditions for optimal expression and immunogenicity of varicella-zoster virus ( VZV ) proteins in a herpes simplex virus-1 ( HSV-1 ) vector , we selected the VZV glycoprotein E ( gE ) , encoded by ORF 68 and the VZV product of ORF 62 , an immediate-early major tegument protein ( IE62 ) .

In order to evaluate the conditions for optimal expression and immunogenicity of <protein>varicella-zoster virus ( VZV ) proteins</protein> in a herpes simplex virus-1 ( HSV-1 ) vector , we selected the <protein>VZV glycoprotein E</protein> ( <protein>gE</protein> ) , encoded by <dna>ORF 68</dna> and the <protein>VZV product</protein> of <dna>ORF 62</dna> , an <protein>immediate-early major tegument protein</protein> ( <protein>IE62</protein> ) .


Three HSV/VZV recombinants were generated : ( 1 ) VZV gE protein coding sequences along with the promoter region were inserted into the thymidine kinase ( TK ) gene of HSV-1 strain KOS ; ( 2 ) VZV gE expressed from the HSV-1 ICP4 promoter was inserted into the glycoprotein C ( gC ) gene of HSV-1 strain F ; and ( 3 ) VZV IE62 protein coding sequences under the control of the HSV-1 ICP4 promoter were inserted into the gC gene of HSV-1 strain F .

Three <dna>HSV/VZV recombinants</dna> were generated : ( 1 ) <dna>VZV gE protein coding sequences</dna> along with the <protein>promoter region</protein> were inserted into the <dna>thymidine kinase ( TK ) gene</dna> of HSV-1 strain KOS ; ( 2 ) <protein>VZV gE</protein> expressed from the <dna>HSV-1 ICP4 promoter</dna> was inserted into the <dna>glycoprotein C ( gC ) gene</dna> of HSV-1 strain F ; and ( 3 ) <dna>VZV IE62 protein coding sequences</dna> under the control of the <dna>HSV-1 ICP4 promoter</dna> were inserted into the <dna>gC gene</dna> of HSV-1 strain F .


Immunoblot analysis and immunoperoxidase staining of infected cell monolayers demonstrated vector expression of VZV proteins .

Immunoblot analysis and immunoperoxidase staining of infected cell monolayers demonstrated vector expression of <protein>VZV proteins</protein> .


Following intracranial inoculation in mice , both VZV gE-HSV ( TK ) and VZV IE62-HSV ( gC ) induced an IgG response against VZV gE or VZV IE62 .

Following intracranial inoculation in mice , both <protein>VZV gE-HSV</protein> ( <protein>TK</protein> ) and <protein>VZV IE62-HSV</protein> ( <protein>gC</protein> ) induced an <protein>IgG</protein> response against <protein>VZV gE</protein> or <protein>VZV IE62</protein> .


When tested in cytotoxicity assays using T-lymphocytes from VZV immune human donors , the range of precursor frequencies for T-lymphocytes that recognized VZV gE or VZV IE62 was similar whether these proteins were expressed by HSV-1 or a vaccinia vector .

When tested in cytotoxicity assays using <cell_type>T-lymphocytes</cell_type> from VZV immune human donors , the range of precursor frequencies for <cell_type>T-lymphocytes</cell_type> that recognized <protein>VZV gE</protein> or <protein>VZV IE62</protein> was similar whether these proteins were expressed by HSV-1 or a vaccinia vector .


These experiments demonstrate that HSV-1 is a competent vector for expression of these VZV proteins and support the feasibility of engineering a combined vaccine for these closely related alpha-herpesviruses .

These experiments demonstrate that HSV-1 is a competent vector for expression of these <protein>VZV proteins</protein> and support the feasibility of engineering a combined vaccine for these closely related alpha-herpesviruses .


Constitutive dephosphorylation and activation of a member of the nuclear factor of activated T cells , NF-AT1 , in Tax-expressing and type I human T-cell leukemia virus-infected human T cells .

Constitutive dephosphorylation and activation of a member of the <protein>nuclear factor of activated T cells</protein> , <protein>NF-AT1</protein> , in <cell_type>Tax-expressing and type I human T-cell leukemia virus-infected human T cells</cell_type> .


The tax gene product of the type I human T-cell leukemia virus ( HTLV-I ) transactivates interleukin-2 ( IL-2 ) gene through activation of an enhancer termed CD28 responsive element ( CD28RE ) .

The tax gene product of the type I human T-cell leukemia virus ( HTLV-I ) transactivates <dna>interleukin-2 ( IL-2 ) gene</dna> through activation of an enhancer termed <dna>CD28 responsive element</dna> ( <dna>CD28RE</dna> ) .


Tax activation of the CD28RE is partially mediated by a member of the nuclear factor of activated T cells , NF-AT1 .

<protein>Tax</protein> activation of the <dna>CD28RE</dna> is partially mediated by a member of the <protein>nuclear factor of activated T cells</protein> , <protein>NF-AT1</protein> .


We have previously shown that NF-AT1 is constitutively active in Jurkat T cells stably transfected with the Tax cDNA , although the underlying molecular mechanism and physiological relevance of this finding remain unclear .

We have previously shown that <protein>NF-AT1</protein> is constitutively active in <cell_line>Jurkat T cells</cell_line> stably transfected with the <dna>Tax cDNA</dna> , although the underlying molecular mechanism and physiological relevance of this finding remain unclear .


In this report , we demonstrate that the active form of NF-AT1 is also present in the nuclei of HTLV-I-transformed T cells that express the Tax protein .

In this report , we demonstrate that the active form of <protein>NF-AT1</protein> is also present in the nuclei of <cell_type>HTLV-I-transformed T cells</cell_type> that express the <protein>Tax protein</protein> .


Interestingly , the constitutive activation of NF-AT1 in these T cells is associated with its dephosphorylation .

Interestingly , the constitutive activation of <protein>NF-AT1</protein> in these <cell_type>T cells</cell_type> is associated with its dephosphorylation .


Furthermore , the dephosphorylated NF-AT1 can be rapidly rephosphorylated when the cells are incubated with cyclosporin A , an immunosuppressant inhibiting the serine/threonine phosphatase calcineurin .

Furthermore , the dephosphorylated <protein>NF-AT1</protein> can be rapidly rephosphorylated when the cells are incubated with cyclosporin A , an immunosuppressant inhibiting the <protein>serine/threonine phosphatase</protein> <protein>calcineurin</protein> .


These results suggest that activation of NF-AT1 in Tax-expressing and HTLV-I-transformed T cells results from its dephosphorylation , which in turn may be due to deregulation of calcineurin

These results suggest that activation of <protein>NF-AT1</protein> in <cell_line>Tax-expressing and HTLV-I-transformed T cells</cell_line> results from its dephosphorylation , which in turn may be due to deregulation of <protein>calcineurin</protein>


Expression of NFAT-family proteins in normal human T cells .

Expression of <protein>NFAT-family proteins</protein> in <cell_type>normal human T cells</cell_type> .


NFAT proteins constitute a family of transcription factors involved in mediating signal transduction .

<protein>NFAT proteins</protein> constitute a family of <protein>transcription factors</protein> involved in mediating signal transduction .


Using a panel of specific antisera in immunoprecipitation assays , we found that NFATp ( 135 kDa ) is constitutively expressed in normal human T cells , while synthesis of NFATc ( predominant form of 86 kDa ) is induced by ionomycin treatment .

Using a panel of specific antisera in immunoprecipitation assays , we found that <protein>NFATp</protein> ( <protein>135 kDa</protein> ) is constitutively expressed in <cell_type>normal human T cells</cell_type> , while synthesis of <protein>NFATc</protein> ( predominant form of <protein>86 kDa</protein> ) is induced by ionomycin treatment .


NFAT4/x was very weakly expressed in unstimulated cells , and its level did not increase upon treatment with activating agents .

<protein>NFAT4/x</protein> was very weakly expressed in unstimulated cells , and its level did not increase upon treatment with activating agents .


NFAT3 protein was not observed under any conditions .

<protein>NFAT3 protein</protein> was not observed under any conditions .


Higher-molecular-weight species of NFATc ( of 110 and 140 kDa ) were also detected .

<protein>Higher-molecular-weight species</protein> of <protein>NFATc</protein> ( of 110 and 140 kDa ) were also detected .


In addition , translation of NFATc mRNA apparently initiates at two different AUG codons , giving rise to proteins that differ in size by 36 amino acids .

In addition , translation of <rna>NFATc mRNA</rna> apparently initiates at two different <dna>AUG codons</dna> , giving rise to proteins that differ in size by 36 amino acids .


Additional size heterogeneity of both NFATc and NFATp results from phosphorylation .

Additional size heterogeneity of both <protein>NFATc</protein> and <protein>NFATp</protein> results from phosphorylation .


In contrast to ionomycin treatment , exposure of cells to phorbol myristate acetate ( PMA ) plus anti-CD28 did not induce NFATc , indicating that under these conditions , interleukin-2 synthesis by these cells is apparently independent of NFATc .

In contrast to ionomycin treatment , exposure of cells to phorbol myristate acetate ( PMA ) plus <protein>anti-CD28</protein> did not induce <protein>NFATc</protein> , indicating that under these conditions , <protein>interleukin-2</protein> synthesis by these cells is apparently independent of <protein>NFATc</protein> .


In DNA binding assays , both PMA plus anti-CD28 and PMA plus ionomycin resulted in nuclear NFAT .

In DNA binding assays , both PMA plus <protein>anti-CD28</protein> and PMA plus ionomycin resulted in <protein>nuclear NFAT</protein> .


Surprisingly , the PMA-ionomycin-induced synthesis of NFATc that was detected by immunoprecipitation was not mirrored in the DNA binding assays : nearly all of the activity was due to NFATp .

Surprisingly , the PMA-ionomycin-induced synthesis of <protein>NFATc</protein> that was detected by immunoprecipitation was not mirrored in the DNA binding assays : nearly all of the activity was due to <protein>NFATp</protein> .


This is the first study of expression of all family members at the protein level in normal human T cells .

This is the first study of expression of all family members at the protein level in <cell_type>normal human T cells</cell_type> .


Of the GATA-binding proteins , only GATA-4 selectively regulates the human IL-5 gene promoter in IL-5 producing cells which express multiple GATA-binding proteins .

Of the <protein>GATA-binding proteins</protein> , only <protein>GATA-4</protein> selectively regulates the <dna>human IL-5 gene promoter</dna> in <cell_type>IL-5 producing cells</cell_type> which express <protein>multiple GATA-binding proteins</protein> .


Interleukin-5 ( IL-5 ) is produced by T lymphocytes and known to support B cell growth and eosinophilic differentiation of the progenitor cells .

<protein>Interleukin-5</protein> ( <protein>IL-5</protein> ) is produced by <cell_type>T lymphocytes</cell_type> and known to support B cell growth and eosinophilic differentiation of the <cell_type>progenitor cells</cell_type> .


Using ATL-16T cells which express IL-5 mRNA , we have identified a region , within the human IL-5 gene promoter , that regulates IL-5 gene transcription .

Using <cell_line>ATL-16T cells</cell_line> which express <rna>IL-5 mRNA</rna> , we have identified a region , within the <dna>human IL-5 gene promoter</dna> , that regulates <dna>IL-5 gene</dna> transcription .


This cis-acting sequence contains the core binding motif , ( A/T ) GATA ( A/G ) , for GATA-binding family proteins and thus suggests the involvement of these family members .

This <dna>cis-acting sequence</dna> contains the <dna>core binding motif</dna> , ( A/T ) GATA ( A/G ) , for <protein>GATA-binding family proteins</protein> and thus suggests the involvement of these family members .


In this report , we describe the cloning of human GATA-4 ( hGATA-4 ) and show that hGATA-4 selectively interacts with the -70 GATA site within the IL-5 proximal promoter region .

In this report , we describe the cloning of <protein>human GATA-4</protein> ( <protein>hGATA-4</protein> ) and show that <protein>hGATA-4</protein> selectively interacts with the <dna>-70 GATA site</dna> within the <dna>IL-5 proximal promoter region</dna> .


By promoter deletion and mutation analyses , we established this region as a positive regulatory element .

By promoter deletion and mutation analyses , we established this region as a <dna>positive regulatory element</dna> .


Cotransfection experiments revealed that both hGATA-4 and PMA/A23187 stimulation are necessary for the IL-5 promoter activation .

Cotransfection experiments revealed that both <protein>hGATA-4</protein> and PMA/A23187 stimulation are necessary for the <dna>IL-5 promoter</dna> activation .


The requirement of another regulatory element called CLE0 , which lies downstream of the -70 GATA site , was also demonstrated .

The requirement of another <dna>regulatory element</dna> called <dna>CLE0</dna> , which lies downstream of the <dna>-70 GATA site</dna> , was also demonstrated .


ATL-16T cells express mRNA of three GATA-binding proteins , hGATA-2 , hGATA-3 and hGATA-4 , and each of them has a potential to bind to the consensus ( A/T ) GATA ( G/ A ) motif .

<cell_line>ATL-16T cells</cell_line> express <rna>mRNA</rna> of three <protein>GATA-binding proteins</protein> , <protein>hGATA-2</protein> , <protein>hGATA-3</protein> and <protein>hGATA-4</protein> , and each of them has a potential to bind to the <dna>consensus ( A/T ) GATA ( G/ A ) motif</dna> .


However , using ATL-16T nuclear extract , we demonstrated that GATA-4 is the only GATA-binding protein that forms specific DNA-protein complex with the -70 GATA site .

However , using ATL-16T nuclear extract , we demonstrated that <protein>GATA-4</protein> is the only <protein>GATA-binding protein</protein> that forms specific <protein>DNA-protein complex</protein> with the <dna>-70 GATA site</dna> .


The electrophoretic mobility shift assay with extracts of COS cells expressing GATA-binding proteins showed that GATA-4 has the highest binding affinity to the -70 GATA site among the three GATA-binding proteins .

The electrophoretic mobility shift assay with extracts of <cell_line>COS cells</cell_line> expressing <protein>GATA-binding proteins</protein> showed that <protein>GATA-4</protein> has the highest binding affinity to the <dna>-70 GATA site</dna> among the three <protein>GATA-binding proteins</protein> .


When the transactivation ability was compared among the three , GATA-4 showed the highest activity .

When the transactivation ability was compared among the three , <protein>GATA-4</protein> showed the highest activity .


These results demonstrate the selective role of GATA-4 in the transcriptional regulation of the IL-5 gene in a circumstance where multiple members of the GATA-binding proteins are expressed .

These results demonstrate the selective role of <protein>GATA-4</protein> in the transcriptional regulation of the <dna>IL-5 gene</dna> in a circumstance where multiple members of the <protein>GATA-binding proteins</protein> are expressed .


Control of NFATx1 nuclear translocation by a calcineurin-regulated inhibitory domain .

Control of <protein>NFATx1</protein> nuclear translocation by a <protein>calcineurin-regulated inhibitory domain</protein> .


The nuclear factor of activated T cells ( NFAT ) regulates cytokine gene expression in T cells through cis-acting elements located in the promoters of several cytokine genes .

The <protein>nuclear factor of activated T cells</protein> ( <protein>NFAT</protein> ) regulates <dna>cytokine gene</dna> expression in <cell_type>T cells</cell_type> through <dna>cis-acting elements</dna> located in the <dna>promoters</dna> of several <dna>cytokine genes</dna> .


NFATx1 , which is preferentially expressed in the thymus and peripheral blood leukocytes , is one of four members of the NFAT family of transcription factors .

<protein>NFATx1</protein> , which is preferentially expressed in the thymus and <cell_type>peripheral blood leukocytes</cell_type> , is one of four members of the <protein>NFAT family</protein> of <protein>transcription factors</protein> .


We have performed domain analysis of NFATx1 by examining the effects of deletion mutations .

We have performed domain analysis of <protein>NFATx1</protein> by examining the effects of deletion mutations .


We found that NFATx1 DNA binding activity and interaction with AP-1 polypeptides were dependent on its central Rel similarity region and that transcriptional activation was reduced by deletions of either its N-terminal domain or its C-terminal domain , suggesting the presence of intrinsic transcriptional activation motifs in both regions .

We found that <protein>NFATx1</protein> DNA binding activity and interaction with AP-1 polypeptides were dependent on its <protein>central Rel similarity region</protein> and that transcriptional activation was reduced by deletions of either its <protein>N-terminal domain</protein> or its <protein>C-terminal domain</protein> , suggesting the presence of <protein>intrinsic transcriptional activation motifs</protein> in both regions .


We also identified a potent inhibitory sequence within its N-terminal domain .

We also identified a potent <protein>inhibitory sequence</protein> within its <protein>N-terminal domain</protein> .


We show that the inactivation of the inhibition was dependent on the activity of calcineurin , a calcium-calmodulin-dependent phosphatase .

We show that the inactivation of the inhibition was dependent on the activity of <protein>calcineurin</protein> , a <protein>calcium-calmodulin-dependent phosphatase</protein> .