Aging has been associated with intrinsic changes of the humoral immune response , which may lead to an increased occurrence of autoimmune disorders and pathogenic susceptibility .


The transcription factor Pax-5 is a key regulator of B cell development .

The <protein>transcription factor</protein> <protein>Pax-5</protein> is a key regulator of B cell development .


Pax-5a/B cell-specific activator protein and an alternatively spliced isoform , Pax-5d , may have opposing functions in transcriptional regulation due to the lack of a transactivation domain in Pax-5d .

<protein>Pax-5a/B cell-specific activator protein</protein> and an <protein>alternatively spliced isoform , Pax-5d</protein> , may have opposing functions in transcriptional regulation due to the lack of a <protein>transactivation domain</protein> in <protein>Pax-5d</protein> .


To study B cell -specific changes that occur during the aging process , we investigated expression patterns of Pax-5a and 5d in mature B cells of young and aged mice .

To study <cell_type>B cell</cell_type> -specific changes that occur during the aging process , we investigated expression patterns of <protein>Pax-5a and 5d</protein> in <cell_type>mature B cells</cell_type> of young and aged mice .


RNase protection assays showed a similar transcriptional pattern for both age groups that indicates that aging has no affect on transcription initiation or alternative splicing for either isoform .

RNase protection assays showed a similar transcriptional pattern for both age groups that indicates that aging has no affect on transcription initiation or alternative splicing for either <protein>isoform</protein> .


In contrast , a significant reduction in the DNA binding activity of Pax-5a but not Pax-5d protein was observed in aged B cells in vitro , while Western blot analyses showed that similar levels of Pax-5a and 5d proteins were present in both age groups .

In contrast , a significant reduction in the DNA binding activity of <protein>Pax-5a</protein> but not <protein>Pax-5d</protein> protein was observed in <cell_type>aged B cells</cell_type> in vitro , while Western blot analyses showed that similar levels of <protein>Pax-5a and 5d</protein> proteins were present in both age groups .


The observed decrease in Pax-5a binding activity correlated with changes in expression of two Pax-5 target genes in aged B cells .

The observed decrease in <protein>Pax-5a</protein> binding activity correlated with changes in expression of two <dna>Pax-5 target genes</dna> in <cell_type>aged B cells</cell_type> .


Expression of the Ig J chain and the secreted form of Ig mu , which are both known to be suppressed by Pax-5a in mature B cells , were increased in B cells of aged mice .

Expression of the <protein>Ig J chain</protein> and the secreted form of <protein>Ig mu</protein> , which are both known to be suppressed by <protein>Pax-5a</protein> in <cell_type>mature B cells</cell_type> , were increased in <cell_type>B cells</cell_type> of aged mice .


Together , our studies suggest that changes associated with the aging phenotype cause posttranslational modification ( s ) of Pax-5a but not Pax-5d , which may lead to an abnormal B cell phenotype in aged mice , associated with elevated levels of J chain , and secretion of IgM

Together , our studies suggest that changes associated with the aging phenotype cause posttranslational modification ( s ) of <protein>Pax-5a</protein> but not <protein>Pax-5d</protein> , which may lead to an abnormal B cell phenotype in aged mice , associated with elevated levels of <protein>J chain</protein> , and secretion of <protein>IgM</protein>


NF-kappa B/Rel participation in the lymphokine-dependent proliferation of T lymphoid cells .

<protein>NF-kappa B/Rel</protein> participation in the lymphokine-dependent proliferation of <cell_type>T lymphoid cells</cell_type> .


Proliferative responses of lymphoid cells to IL-2 and IL-4 depend on activation of the cells , but the mechanism ( s ) by which activation enhances cellular competence to respond to cytokines is not fully understood .

Proliferative responses of <cell_type>lymphoid cells</cell_type> to <protein>IL-2</protein> and <protein>IL-4</protein> depend on activation of the cells , but the mechanism ( s ) by which activation enhances cellular competence to respond to <protein>cytokines</protein> is not fully understood .


The NF-kappaB/Rel family represents one signal transduction pathway induced during such activation .

The <protein>NF-kappaB/Rel family</protein> represents one signal transduction pathway induced during such activation .


We show in this study that inhibition of NF-kappaB through the expression of an IkappaBalpha ( inhibitory protein that dissociates from NF-kappaB ) mutant refractory to signal-induced degradation ( IkappaBalpha ( DeltaN ) ) interfered with the acquisition of competence to proliferate in response to IL-4 as well as IL-2 .

We show in this study that inhibition of <protein>NF-kappaB</protein> through the expression of an <protein>IkappaBalpha</protein> ( <protein>inhibitory protein</protein> that dissociates from <protein>NF-kappaB</protein> ) mutant refractory to signal-induced degradation ( <protein>IkappaBalpha ( DeltaN )</protein> ) interfered with the acquisition of competence to proliferate in response to <protein>IL-4</protein> as well as <protein>IL-2</protein> .


Thymocytes and T cells from IkappaBalpha ( DeltaN ) transgenic mice expressed normal levels of IL-2R subunits .

<cell_type>Thymocytes</cell_type> and <cell_type>T cells</cell_type> from <protein>IkappaBalpha ( DeltaN )</protein> transgenic mice expressed normal levels of <protein>IL-2R subunits</protein> .


However , transgenic cells exhibited a dramatic defect in Stat5A activation treatment with IL-2 , and a similar defect was observed for IL-4 -induced Stat5 .

However , <cell_type>transgenic cells</cell_type> exhibited a dramatic defect in Stat5A activation treatment with <protein>IL-2</protein> , and a similar defect was observed for <protein>IL-4</protein> -induced <protein>Stat5</protein> .


In contrast , T lymphoid cells with inhibition of NF-kappaB showed normal insulin receptor substrate-2 phosphorylation and only a modest decrease in Stat6 activation and insulin receptor substrate-1 phosphorylation after IL-4 stimulation .

In contrast , <cell_type>T lymphoid cells</cell_type> with inhibition of <protein>NF-kappaB</protein> showed normal insulin receptor substrate-2 phosphorylation and only a modest decrease in Stat6 activation and insulin receptor substrate-1 phosphorylation after IL-4 stimulation .


These results indicate that the NF-kappaB/Rel/IkappaBalpha system can regulate cytokine receptor capacitation through effects on the induction of downstream signaling by the Stat transcription factor family .

These results indicate that the NF-kappaB/Rel/IkappaBalpha system can regulate <protein>cytokine receptor</protein> capacitation through effects on the induction of downstream signaling by the <protein>Stat transcription factor family</protein> .


BMS-189453 , a novel retinoid receptor antagonist , is a potent testicular toxin .


BMS-189453 is a synthetic retinoid that acts as an antagonist at retinoic acid receptors alpha , beta , and gamma .

BMS-189453 is a synthetic retinoid that acts as an antagonist at <protein>retinoic acid receptors alpha , beta , and gamma</protein> .


In Sprague Dawley rats at daily oral doses of 15 , 60 , or 240 mg/kg for 1 month , BMS-189453 produced increases in leukocyte counts , alkaline phosphatase and alanine aminotransferase levels , and marked testicular degeneration and atrophy at all doses .

In Sprague Dawley rats at daily oral doses of 15 , 60 , or 240 mg/kg for 1 month , BMS-189453 produced increases in <cell_type>leukocyte</cell_type> counts , <protein>alkaline phosphatase</protein> and <protein>alanine aminotransferase</protein> levels , and marked testicular degeneration and atrophy at all doses .


Significant overt signs of toxicity and deaths occurred at 240 mg/kg , whereas body-weight and food-consumption decreases occurred at 60 and 240 mg/kg .


When BMS-189453 was administered to male rats at daily doses ranging from 12.5 to 100 mg/kg for 1 week , only minimal testicular changes occurred at all doses , shortly after the dosing period .


However , after a 1-month drug-free observation period , marked testicular atrophy was evident at all doses .


BMS-189453 was then administered at doses of 2 , 10 , or 50 mg/kg to male rats for 1 , 3 , or 7 consecutive days .


Dose- and duration-dependent testicular toxicity that occurred after a 1-month observation period did not recover , and , in some cases , was more severe 4 months after the last dose .


In rabbits administered BMS-189453 at oral doses of 2 , 10 , or 50 mg/kg for 1 week , testicular degeneration and atrophy were evident in the high-dose group at 1 month following treatment .


These studies indicate that retinoid antagonists can selectively produce progressive and prolonged testicular toxicity after single or repeated oral doses that are otherwise well tolerated .


Sequential involvement of NFAT and Egr transcription factors in FasL regulation .

Sequential involvement of <protein>NFAT</protein> and <protein>Egr</protein> <protein>transcription factors</protein> in FasL regulation .


The critical function of NFAT proteins in maintaining lymphoid homeostasis was revealed in mice lacking both NFATp and NFAT4 ( DKO ) .

The critical function of <protein>NFAT proteins</protein> in maintaining lymphoid homeostasis was revealed in mice lacking both <protein>NFATp</protein> and <protein>NFAT4</protein> ( DKO ) .


DKO mice exhibit increased lymphoproliferation , decreased activation-induced cell death , and impaired induction of FasL .

DKO mice exhibit increased lymphoproliferation , decreased activation-induced cell death , and impaired induction of <protein>FasL</protein> .


The transcription factors Egr2 and Egr3 are potent activators of FasL expression .

The <protein>transcription factors</protein> <protein>Egr2</protein> and <protein>Egr3</protein> are potent activators of FasL expression .


Here we find that Egr2 and Egr3 are NFAT target genes .

Here we find that <dna>Egr2</dna> and <dna>Egr3</dna> are <dna>NFAT target genes</dna> .


Activation of FasL occurs via the NFAT -dependent induction of Egr3 , as demonstrated by the ability of exogenously provided NFATp to restore Egr -dependent FasL promoter activity in DKO lymph node cells .

Activation of <protein>FasL</protein> occurs via the <protein>NFAT</protein> -dependent induction of <dna>Egr3</dna> , as demonstrated by the ability of exogenously provided <protein>NFATp</protein> to restore <protein>Egr</protein> -dependent FasL promoter activity in <cell_type>DKO lymph node cells</cell_type> .


Further , Egr3 expression is enriched in Th1 cells , suggesting a molecular basis for the known preferential expression of FasL in the Th1 versus Th2 subset .

Further , <dna>Egr3</dna> expression is enriched in <cell_type>Th1 cells</cell_type> , suggesting a molecular basis for the known preferential expression of <protein>FasL</protein> in the Th1 versus Th2 subset .


Mechanisms and clinical relevance of nongenomic glucocorticoid actions .


Glucocorticoids have profound anti-inflammatory and immunosuppressive actions when used therapeutically .


The therapeutic dose is quite variable and depends on the disease , but ranges from very low to extremely high .


The rationale for the use of various dosage regimens for specific clinical indications is the existence of three distinct , therapeutically relevant effects : genomic , specific nongenomic and unspecific nongenomic .


Genomic effects are mediated by cytosolic receptors that alter expression of specific genes .

Genomic effects are mediated by <protein>cytosolic receptors</protein> that alter expression of specific genes .


Specific nongenomic effects occur within a few minutes and are mediated by steroid-selective membrane receptors .

Specific nongenomic effects occur within a few minutes and are mediated by <protein>steroid-selective membrane receptors</protein> .


Unspecific nongenomic effects occur within seconds , but only at high glucocorticoid dosages , and seem to result from direct interactions with biological membranes .


For unspecific nongenomic effects , methylprednisolone and other glucocorticoids have been shown to inhibit cation cycling across the plasma membrane , but to have little effect on protein synthesis .


Thus , glucocorticoids could diminish or prevent the acute immune response by interfering with processes such as the rise in intracellular Ca2+ concentration .


It is proposed that the additional therapeutic benefit of higher doses is obtained via these nongenomic effects .


High glucose-induced intercellular adhesion molecule-1 ( ICAM-1 ) expression through an osmotic effect in rat mesangial cells is PKC-NF-kappa B-dependent .

High glucose-induced <protein>intercellular adhesion molecule-1</protein> ( <protein>ICAM-1</protein> ) expression through an osmotic effect in <cell_type>rat mesangial cells</cell_type> is PKC-NF-kappa B-dependent .


AIMS/HYPOTHESIS : Infiltration of mononuclear cells and glomerular enlargement accompanied by glomerular cell proliferation are very early characteristics of the pathophysiology of diabetes .

AIMS/HYPOTHESIS : Infiltration of <cell_type>mononuclear cells</cell_type> and glomerular enlargement accompanied by <cell_type>glomerular cell</cell_type> proliferation are very early characteristics of the pathophysiology of diabetes .


To clarify the mechanism of early diabetic nephropathy , we measured [ 3H ] -thymidine incorporation and cell numbers to show the influence of a high ambient glucose concentration and the osmotic effect on rat mesangial cell proliferation .

To clarify the mechanism of early diabetic nephropathy , we measured [ 3H ] -thymidine incorporation and cell numbers to show the influence of a high ambient glucose concentration and the osmotic effect on <cell_type>rat mesangial cell</cell_type> proliferation .


We also measured the effect of high glucose on the expression of intercellular adhesion molecule-1 and vascular adhesion molecule-1 by flow cytometry and semiquantitative RT-PCR in mesangial cells and the adhesion of leukocytes to mesangial cells .

We also measured the effect of high glucose on the expression of <protein>intercellular adhesion molecule-1</protein> and <protein>vascular adhesion molecule-1</protein> by flow cytometry and semiquantitative RT-PCR in <cell_type>mesangial cells</cell_type> and the adhesion of <cell_type>leukocytes</cell_type> to <cell_type>mesangial cells</cell_type> .


METHODS/RESULTS : Cells exposed to high D-glucose ( 30 mmol/l ) caused an increase in [ 3H ] -thymidine incorporation and cell numbers at 24 and 48 h and normalized at 72 h ( p < 0.05 ) , whereas these changes were not found in high mannitol ( 30 mmol/l ) , IL-1 beta , or TNF alpha -stimulated mesangial cells .

METHODS/RESULTS : Cells exposed to high D-glucose ( 30 mmol/l ) caused an increase in [ 3H ] -thymidine incorporation and cell numbers at 24 and 48 h and normalized at 72 h ( p < 0.05 ) , whereas these changes were not found in high mannitol ( 30 mmol/l ) , <protein>IL-1 beta</protein> , or <protein>TNF alpha</protein> -stimulated <cell_type>mesangial cells</cell_type> .


Cells exposed to high-glucose ( 15 , 30 , or 60 mmol/l ) or osmotic agents ( L-glucose , raffinose and mannitol ) showed that intercellular adhesion molecule-1 expression began to increase after 24 h , reached its maximum at 24 and 48 h and gradually decreased afterwards .

Cells exposed to high-glucose ( 15 , 30 , or 60 mmol/l ) or osmotic agents ( L-glucose , raffinose and mannitol ) showed that <protein>intercellular adhesion molecule-1</protein> expression began to increase after 24 h , reached its maximum at 24 and 48 h and gradually decreased afterwards .


The stimulatory effects of high glucose and high mannitol on mRNA expression were observed as early as 6 h and reached its maximum at 12 h .


Up-regulation of ICAM-1 protein and mRNA was also found in IL-1-beta and TNF-alpha -stimulated mesangial cells .

Up-regulation of <protein>ICAM-1</protein> protein and mRNA was also found in <protein>IL-1-beta</protein> and <protein>TNF-alpha</protein> -stimulated <cell_type>mesangial cells</cell_type> .


Neither vascular adhesion molecule-1 protein nor mRNA expression was , however , affected by high glucose and high mannitol .

Neither <protein>vascular adhesion molecule-1</protein> protein nor mRNA expression was , however , affected by high glucose and high mannitol .


Notably , the protein kinase C inhibitors calphostin C and staurosporine reduced high glucose- or high mannitol-induced intercellular adhesion molecule-1 mRNA expression and high glucose-induced proliferation .

Notably , the protein kinase C inhibitors calphostin C and staurosporine reduced high glucose- or high mannitol-induced <rna>intercellular adhesion molecule-1</rna> mRNA expression and high glucose-induced proliferation .


Furthermore , the NF-kappa B inhibitor N-tosyl-L-phenylalanine chloromethyl ketone reduced high glucose- or high mannitol-induced intercellular adhesion molecule-1 mRNA expression and high glucose-induced proliferation .

Furthermore , the NF-kappa B inhibitor N-tosyl-L-phenylalanine chloromethyl ketone reduced high glucose- or high mannitol-induced <rna>intercellular adhesion molecule-1</rna> mRNA expression and high glucose-induced proliferation .


Results showed that high glucose ( 15 , 30 mmol/l ) or high concentrations of osmotic agents remarkably increased the number of adherent leukocytes to mesangial cells ( p < 0.01 ) compared with control cells ( 5 mmol/l D-glucose ) .

Results showed that high glucose ( 15 , 30 mmol/l ) or high concentrations of osmotic agents remarkably increased the number of adherent <cell_type>leukocytes</cell_type> to <cell_type>mesangial cells</cell_type> ( p < 0.01 ) compared with <cell_type>control cells</cell_type> ( 5 mmol/l D-glucose ) .


Functional blocking of intercellular adhesion molecule-1 on mesangial cells with rat intercellular adhesion molecule-1 monoclonal antibody , calphostin C , staurosporine , or N-tosyl-L-phenylalanine chloromethyl ketone significantly inhibited high glucose- or high mannitol-induced increase in leukocyte adhesion ( p < < 0.05 ) .

Functional blocking of <protein>intercellular adhesion molecule-1</protein> on <cell_type>mesangial cells</cell_type> with <protein>rat intercellular adhesion molecule-1 monoclonal antibody</protein> , calphostin C , staurosporine , or N-tosyl-L-phenylalanine chloromethyl ketone significantly inhibited high glucose- or high mannitol-induced increase in leukocyte adhesion ( p < < 0.05 ) .


CONCLUSION/INTERPRETATION : These results suggest that high glucose can upregulate intercellular adhesion molecule-1 protein and mRNA expression but not vascular adhesion molecule-1 expression in mesangial cells and promote leukocyte adhesion through up-regulation of intercellular adhesion molecule-1 through osmotic effect , possibly depending on the protein kinase C nuclear factor-kappa B ( PKC - NF-kappa B ) pathway .

CONCLUSION/INTERPRETATION : These results suggest that high glucose can upregulate <protein>intercellular adhesion molecule-1</protein> protein and mRNA expression but not vascular adhesion molecule-1 expression in <cell_type>mesangial cells</cell_type> and promote leukocyte adhesion through up-regulation of <protein>intercellular adhesion molecule-1</protein> through osmotic effect , possibly depending on the <protein>protein kinase C</protein> <protein>nuclear factor-kappa B</protein> ( <protein>PKC</protein> - <protein>NF-kappa B</protein> ) pathway .


High glucose itself can also promote mesangial cell proliferation through the PKC-NF-kappa B pathways .


We conclude that hyperglycaemia in itself seems to be an important factor in the development of early diabetic nephropathy .


Combined corticosteroid/ granulocyte colony-stimulating factor ( G-CSF ) therapy in the treatment of severe congenital neutropenia unresponsive to G-CSF : Activated glucocorticoid receptors synergize with G-CSF signals .

Combined corticosteroid/ <protein>granulocyte colony-stimulating factor</protein> ( <protein>G-CSF</protein> ) therapy in the treatment of severe congenital neutropenia unresponsive to <protein>G-CSF</protein> : Activated <protein>glucocorticoid receptors</protein> synergize with <protein>G-CSF</protein> signals .


More than 90 % of patients with severe congenital neutropenia ( SCN ) respond to granulocyte colony-stimulating factor ( G-CSF ) therapy .

More than 90 % of patients with severe congenital neutropenia ( SCN ) respond to <protein>granulocyte colony-stimulating factor</protein> ( <protein>G-CSF</protein> ) therapy .


The basis for the refractory state in the remaining patients is unknown .


To address this issue , we studied a child with SCN who was totally unresponsive to G-CSF and had a novel point mutation in the extracellular domain of the G-CSF receptor ( GCSF-R ) .

To address this issue , we studied a child with SCN who was totally unresponsive to <protein>G-CSF</protein> and had a novel point mutation in the <protein>extracellular domain</protein> of the <protein>G-CSF receptor</protein> ( <protein>GCSF-R</protein> ) .


Marrow stromal support of granulopoiesis was evaluated by plating CD34 ( + ) cells on preformed stromal layers .

Marrow stromal support of granulopoiesis was evaluated by plating <cell_type>CD34 ( + ) cells</cell_type> on preformed stromal layers .


Nonadherent cells were harvested and assayed in clonogenic assays for granulocytic colony production .

<cell_type>Nonadherent cells</cell_type> were harvested and assayed in clonogenic assays for granulocytic colony production .


The in vitro effect of G-CSF and corticosteroids on granulopoiesis was evaluated in clonogenic assays of marrow mononuclear cells , by proliferation studies of the murine myeloid cell line 32D expressing the patient 's mutated G-CSFR , and by measuring STAT5 activation in nuclear extracts from stimulated cells .

The in vitro effect of <protein>G-CSF</protein> and corticosteroids on granulopoiesis was evaluated in clonogenic assays of <cell_type>marrow mononuclear cells</cell_type> , by proliferation studies of the <cell_line>murine myeloid cell line 32D</cell_line> expressing the patient 's <protein>mutated G-CSFR</protein> , and by measuring STAT5 activation in nuclear extracts from <cell_type>stimulated cells</cell_type> .


Patient 's stroma supported granulopoiesis derived from control marrow CD34 ( + ) cells in a normal manner .

Patient 's stroma supported granulopoiesis derived from control <cell_type>marrow CD34 ( + ) cells</cell_type> in a normal manner .


Normal stroma , however , failed to induce granulopoiesis from patient 's CD34 ( + ) cells .

Normal stroma , however , failed to induce granulopoiesis from patient 's <cell_type>CD34 ( + ) cells</cell_type> .


Clonogenic assays of the patient 's marrow mononuclear cells incorporating either G-CSF or hydrocortisone produced little neutrophil growth .

Clonogenic assays of the patient 's <cell_type>marrow mononuclear cells</cell_type> incorporating either <protein>G-CSF</protein> or hydrocortisone produced little neutrophil growth .


In contrast , inclusion of both G-CSF and hydrocortisone in the cytokine `` cocktail '' markedly increased the neutrophil numbers .

In contrast , inclusion of both <protein>G-CSF</protein> and hydrocortisone in the <protein>cytokine `` cocktail ''</protein> markedly increased the neutrophil numbers .


Proliferation of 32D cells expressing the mutated receptor and STAT5 activation were improved by a combination of G-CSF and dexamethasone .

Proliferation of <cell_line>32D cells</cell_line> expressing the <protein>mutated receptor</protein> and STAT5 activation were improved by a combination of <protein>G-CSF</protein> and dexamethasone .


When small daily doses of oral prednisone were then administered to the patient with conventional doses of subcutaneous G-CSF , the patient responded with increased neutrophil numbers and with a complete reversal of the infectious problems .

When small daily doses of oral prednisone were then administered to the patient with conventional doses of <protein>subcutaneous G-CSF</protein> , the patient responded with increased neutrophil numbers and with a complete reversal of the infectious problems .


These data provide insight into SCN unresponsive to standard G-CSF treatment and to the potential corrective action of combined treatment with G-CSF and corticosteroids through synergistic activation of STAT5 .

These data provide insight into SCN unresponsive to standard <protein>G-CSF</protein> treatment and to the potential corrective action of combined treatment with <protein>G-CSF</protein> and corticosteroids through synergistic activation of <protein>STAT5</protein> .


Human renal mesangial cells are a target for the anti-inflammatory action of 9-cis retinoic acid .

<cell_type>Human renal mesangial cells</cell_type> are a target for the anti-inflammatory action of 9-cis retinoic acid .


Mesangial cells play an active role in the inflammatory response to glomerular injury .

<cell_type>Mesangial cells</cell_type> play an active role in the inflammatory response to glomerular injury .


We have studied in cultured human mesangial cells ( CHMC ) several effects of 9-cis retinoic acid ( 9-cRA ) , an activator of both retinoic acid receptors ( RARs ) and retinoid X receptors ( RXRs ) .

We have studied in <cell_line>cultured human mesangial cells</cell_line> ( <cell_line>CHMC</cell_line> ) several effects of 9-cis retinoic acid ( 9-cRA ) , an activator of both <protein>retinoic acid receptors</protein> ( <protein>RARs</protein> ) and <protein>retinoid X receptors</protein> ( <protein>RXRs</protein> ) .


9-cRA inhibited foetal calf serum-induced CHMC proliferation .

9-cRA inhibited foetal calf serum-induced <cell_line>CHMC</cell_line> proliferation .


It also prevented CHMC death induced by the inflammatory mediator H ( 2 ) O ( 2 ) .

It also prevented <cell_line>CHMC</cell_line> death induced by the inflammatory mediator H ( 2 ) O ( 2 ) .


This preventive effect was not due to any increase in H ( 2 ) O ( 2 ) catabolism and it persisted even when both catalase and glutathione synthesis were inhibited .


Finally , 9-cRA diminished monocyte adhesion to FCS-stimulated CHMC .

Finally , 9-cRA diminished monocyte adhesion to <cell_line>FCS-stimulated CHMC</cell_line> .


Interestingly , the retinoid also inhibited in FCS-stimulated cells the protein expression of two mesangial adhesion molecules , fibronectin and osteopontin , but it did not modify the protein expression of intercellular adhesion molecule-1 and vascular adhesion molecule-1 .

Interestingly , the retinoid also inhibited in <cell_type>FCS-stimulated cells</cell_type> the protein expression of two <protein>mesangial adhesion molecules</protein> , <protein>fibronectin</protein> and <protein>osteopontin</protein> , but it did not modify the protein expression of <protein>intercellular adhesion molecule-1</protein> and <protein>vascular adhesion molecule-1</protein> .


All major RARs and RXRs isotypes were expressed in CHMC regardless of the presence or absence of 9-cRA .

All major <protein>RARs and RXRs isotypes</protein> were expressed in <cell_line>CHMC</cell_line> regardless of the presence or absence of 9-cRA .


Transcripts to RAR-alpha , RAR-beta and RXR-alpha increased after incubation with 9-cRA whereas RXR-gamma was inhibited , suggesting a major role for RARs and RXRs in 9-cRA-anti-inflammatory effects .

Transcripts to <rna>RAR-alpha</rna> , <rna>RAR-beta</rna> and <rna>RXR-alpha</rna> increased after incubation with 9-cRA whereas <rna>RXR-gamma</rna> was inhibited , suggesting a major role for <protein>RARs</protein> and <protein>RXRs</protein> in 9-cRA-anti-inflammatory effects .


9-cRA was toxic only at 50 microM ( a concentration 50 - 5000 times higher than required for the effects above ) .


Cell death occurred by apoptosis , whose onset was associated with a pronounced increase in catalase activity and reduced glutathione content , being more effectively induced by all-trans retinoic acid .


Modulation of the oxidant/antioxidant balance failed to inhibit apoptosis .


We conclude that mesangial cells might be a target for the treatment of inflammatory glomerulopathies with 9-cRA .

We conclude that <cell_type>mesangial cells</cell_type> might be a target for the treatment of inflammatory glomerulopathies with 9-cRA .


Expression of mammalian defensin genes .

Expression of <dna>mammalian defensin genes</dna> .


Antimicrobial peptides are a prevalent mechanism of host defense found throughout nature .


In mammals , defensins are among the most abundant of these broad-spectrum antibiotics , and are expressed in epithelial and hematopoietic cells .

In mammals , <protein>defensins</protein> are among the most abundant of these broad-spectrum antibiotics , and are expressed in <cell_type>epithelial and hematopoietic cells</cell_type> .


The defensin peptides are especially abundant in neutrophils ; however , gene expression is limited to the promyelocyte stage .

The defensin peptides are especially abundant in <cell_type>neutrophils</cell_type> ; however , gene expression is limited to the promyelocyte stage .


In epithelial cells , defensin genes are found as both constitutively expressed and inducible .

In <cell_type>epithelial cells</cell_type> , <dna>defensin genes</dna> are found as both constitutively expressed and inducible .


Induction has been observed in vitro by stimulation with bacterial lipopolysaccharide as well as inflammatory mediators .


In vivo , up-regulation of several defensin genes occurs in both infectious and inflammatory states .

In vivo , up-regulation of several <dna>defensin genes</dna> occurs in both infectious and inflammatory states .


Gene regulation occurs via signal transduction pathways common to other innate immune responses , utilizing transcription factors such as nuclear factor ( NF ) -kappaB and NF interleukin-6 .

Gene regulation occurs via signal transduction pathways common to other innate immune responses , utilizing <protein>transcription factors</protein> such as <protein>nuclear factor ( NF ) -kappaB</protein> and <protein>NF interleukin-6</protein> .


Together , the data suggest a broad-based innate host defense whereby potent antimicrobial peptides are present to prevent initial colonization by pathogenic microorganisms .


In addition , the recognition of bacteria coupled with a nascent inflammatory response can bolster this defense by a coordinated up-regulation of the peptides .


Multiple signals required for cyclic AMP-responsive element binding protein ( CREB ) binding protein interaction induced by CD3/CD28 costimulation .

Multiple signals required for <protein>cyclic AMP-responsive element binding protein</protein> ( <protein>CREB</protein> ) binding protein interaction induced by CD3/CD28 costimulation .


The optimal activation of cAMP-responsive element binding protein ( CREB ) , similar to the full activation of T lymphocytes , requires the stimulation of both CD3 and CD28 .

The optimal activation of <protein>cAMP-responsive element binding protein</protein> ( <protein>CREB</protein> ) , similar to the full activation of <cell_type>T lymphocytes</cell_type> , requires the stimulation of both <protein>CD3</protein> and <protein>CD28</protein> .


Using a reporter system to detect interaction of CREB and CREB-binding protein ( CBP ) , in this study we found that CREB binds to CBP only by engagement of both CD3 and CD28 .

Using a reporter system to detect interaction of <protein>CREB</protein> and <protein>CREB-binding protein</protein> ( <protein>CBP</protein> ) , in this study we found that <protein>CREB</protein> binds to <protein>CBP</protein> only by engagement of both <protein>CD3</protein> and <protein>CD28</protein> .


CD3/CD28 -promoted CREB - CBP interaction was dependent on p38 mitogen-activated protein kinase ( MAPK ) and calcium/calmodulin-dependent protein kinase ( CaMK ) IV in addition to the previously identified extracellular signal-regulated kinase pathway .

<protein>CD3/CD28</protein> -promoted <protein>CREB</protein> - <protein>CBP</protein> interaction was dependent on <protein>p38 mitogen-activated protein kinase</protein> ( <protein>MAPK</protein> ) and <protein>calcium/calmodulin-dependent protein kinase ( CaMK ) IV</protein> in addition to the previously identified extracellular signal-regulated kinase pathway .


Extracellular signal-regulated kinase , CaMKIV , and p38 MAPK were also the kinases involved in CREB Ser ( 133 ) phosphorylation induced by CD3/CD28 .

<protein>Extracellular signal-regulated kinase , CaMKIV</protein> , and <protein>p38 MAPK</protein> were also the kinases involved in <protein>CREB</protein> Ser ( 133 ) phosphorylation induced by <protein>CD3/CD28</protein> .


A reconstitution experiment illustrated that optimum CREB - CBP interaction and CREB trans-activation were attained when these three kinase pathways were simultaneously activated in T cells .

A reconstitution experiment illustrated that optimum <protein>CREB</protein> - <protein>CBP</protein> interaction and <protein>CREB</protein> trans-activation were attained when these three kinase pathways were simultaneously activated in <cell_type>T cells</cell_type> .


Our results demonstrate that coordinated activation of different kinases leads to full activation of CREB .

Our results demonstrate that coordinated activation of different <protein>kinases</protein> leads to full activation of <protein>CREB</protein> .


Notably , CD28 ligation activated p38 MAPK and CaMKIV , the kinases stimulated by CD3 engagement , suggesting that CD28 acts by increasing the activation extent of p38 MAPK and CaMKIV .

Notably , CD28 ligation activated <protein>p38 MAPK</protein> and <protein>CaMKIV</protein> , the <protein>kinases</protein> stimulated by CD3 engagement , suggesting that <protein>CD28</protein> acts by increasing the activation extent of <protein>p38 MAPK</protein> and <protein>CaMKIV</protein> .


These results support the model of a minimum activation threshold for CREB - CBP interaction that can be reached only when both CD3 and CD28 are stimulated .

These results support the model of a minimum activation threshold for <protein>CREB</protein> - <protein>CBP</protein> interaction that can be reached only when both <protein>CD3</protein> and <protein>CD28</protein> are stimulated .


The murine IL-2 promoter contains distal regulatory elements responsive to the Ah receptor , a member of the evolutionarily conserved bHLH-PAS transcription factor family .

The <dna>murine IL-2 promoter</dna> contains <dna>distal regulatory elements</dna> responsive to the <protein>Ah receptor</protein> , a member of the evolutionarily conserved <protein>bHLH-PAS transcription factor family</protein> .


Signaling through the TCR and costimulatory signals primarily control transcription of the IL-2 gene in naive T cells .

Signaling through the <protein>TCR</protein> and costimulatory signals primarily control transcription of the <dna>IL-2 gene</dna> in <cell_type>naive T cells</cell_type> .


The minimal promoter necessary for this expression lies proximal , between -300 and the transcription start site .

The <dna>minimal promoter</dna> necessary for this expression lies proximal , between -300 and the <dna>transcription start site</dna> .


We had previously shown that activation of the arylhydrocarbon receptor ( AHR ) , a member of the bHLH-PAS family of transcription factors , leads to increased mRNA expression of IL-2 in murine fetal thymocytes .

We had previously shown that activation of the <protein>arylhydrocarbon receptor</protein> ( <protein>AHR</protein> ) , a member of the <protein>bHLH-PAS family of transcription factors</protein> , leads to increased mRNA expression of <rna>IL-2</rna> in <cell_type>murine fetal thymocytes</cell_type> .


The AHR is abundant in the thymus and may play a role for the development of the immune system .

The <protein>AHR</protein> is abundant in the thymus and may play a role for the development of the immune system .


Moreover , its overactivation by chemicals such as dioxins leads to immunosuppression and thymic involution .


Binding motifs for the liganded AHR can be identified in the distal region -1300 to -800 of the mouse IL-2 promoter .

Binding motifs for the <protein>liganded AHR</protein> can be identified in the <dna>distal region -1300 to -800</dna> of the <dna>mouse IL-2 promoter</dna> .


We show here that these DNA motifs , the so-called dioxin response elements , after binding to the liganded AHR are sufficient to transactivate luciferase expression in a reporter gene system .

We show here that these <dna>DNA motifs</dna> , the so-called <dna>dioxin response elements</dna> , after binding to the <protein>liganded AHR</protein> are sufficient to transactivate luciferase expression in a reporter gene system .


The IL-2 gene can be induced by the AHR also in thymocytes in vivo after injection of 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin , a potent ligand of the AHR .

The <dna>IL-2 gene</dna> can be induced by the <protein>AHR</protein> also in <cell_type>thymocytes</cell_type> in vivo after injection of 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin , a potent ligand of the <protein>AHR</protein> .


The AHR mediates the IL-2 induction as shown with AHR -deficient mice .

The <protein>AHR</protein> mediates the IL-2 induction as shown with <protein>AHR</protein> -deficient mice .


However , in spleen cells in vitro costimulation via the TCR is necessary for optimal IL-2 gene induction .

However , in <cell_type>spleen cells</cell_type> in vitro costimulation via the <protein>TCR</protein> is necessary for optimal <dna>IL-2 gene</dna> induction .


Thus , the IL-2 promoter region contains novel distal regulatory elements that can be addressed by the AHR to induce IL-2 and can cooperate with the proximal promoter

Thus , the <dna>IL-2 promoter region</dna> contains novel <dna>distal regulatory elements</dna> that can be addressed by the <protein>AHR</protein> to induce <dna>IL-2</dna> and can cooperate with the <dna>proximal promoter</dna>


Visualization of Syk-antigen receptor interactions using green fluorescent protein : differential roles for Syk and Lyn in the regulation of receptor capping and internalization .

Visualization of <protein>Syk-antigen receptor</protein> interactions using <protein>green fluorescent protein</protein> : differential roles for <protein>Syk</protein> and <protein>Lyn</protein> in the regulation of receptor capping and internalization .


The cross-linking of the B cell Ag receptor ( BCR ) is coupled to the stimulation of multiple intracellular signal transduction cascades via receptor-associated , protein tyrosine kinases of both the Src and Syk families .

The cross-linking of the <protein>B cell Ag receptor</protein> ( <protein>BCR</protein> ) is coupled to the stimulation of multiple intracellular signal transduction cascades via receptor-associated , <protein>protein tyrosine kinases</protein> of both the <protein>Src and Syk families</protein> .


To monitor changes in the subcellular distribution of Syk in B cells responding to BCR cross-linking , we expressed in Syk -deficient DT40 B cells a fusion protein consisting of Syk coupled to green fluorescent protein .

To monitor changes in the subcellular distribution of <protein>Syk</protein> in <cell_type>B cells</cell_type> responding to <protein>BCR</protein> cross-linking , we expressed in <protein>Syk</protein> -deficient <cell_line>DT40 B cells</cell_line> a <protein>fusion protein</protein> consisting of <protein>Syk</protein> coupled to <protein>green fluorescent protein</protein> .


Treatment of these cells with anti-IgM Abs leads to the recruitment of the kinase from cytoplasmic and nuclear compartments to the site of the cross-linked receptor at the plasma membrane .

Treatment of these cells with <protein>anti-IgM Abs</protein> leads to the recruitment of the <protein>kinase</protein> from cytoplasmic and nuclear compartments to the site of the <protein>cross-linked receptor</protein> at the plasma membrane .


The Syk-receptor complexes aggregate into membrane patches that redistribute to form a cap at one pole of the cell .

The <protein>Syk-receptor complexes</protein> aggregate into membrane patches that redistribute to form a cap at one pole of the cell .


Syk is not demonstrably associated with the internalized receptor .

<protein>Syk</protein> is not demonstrably associated with the <protein>internalized receptor</protein> .


Catalytically active Syk promotes and stabilizes the formation of tightly capped BCR complexes at the plasma membrane .

Catalytically active <protein>Syk</protein> promotes and stabilizes the formation of <protein>tightly capped BCR complexes</protein> at the plasma membrane .


Lyn is not required for the recruitment of Syk to the cross-linked receptor , but is required for the internalization of the clustered BCR complexes .

<protein>Lyn</protein> is not required for the recruitment of <protein>Syk</protein> to the <protein>cross-linked receptor</protein> , but is required for the internalization of the clustered <protein>BCR complexes</protein> .


In the absence of Lyn , receptor-Syk complexes at the plasma membrane are long lived , and the receptor-mediated activation of the NF-AT transcription factor is enhanced .

In the absence of <protein>Lyn</protein> , <protein>receptor-Syk complexes</protein> at the plasma membrane are long lived , and the receptor-mediated activation of the <protein>NF-AT transcription factor</protein> is enhanced .


Thus , Lyn appears to function to negatively regulate aspects of BCR -dependent signaling by stimulating receptor internalization and down-regulation .

Thus , <protein>Lyn</protein> appears to function to negatively regulate aspects of <protein>BCR</protein> -dependent signaling by stimulating receptor internalization and down-regulation .


Renal interstitial fibrosis is reduced in angiotensin II type 1a receptor -deficient mice .

Renal interstitial fibrosis is reduced in <protein>angiotensin II type 1a receptor</protein> -deficient mice .


Unilateral ureteral obstruction ( UUO ) results in tubulointerstitial fibrosis of the affected kidney by stimulating the renin-angiotensin system .


This study established a UUO model in angiotensin type 1a receptor ( AT1a ) deficient ( mutant ) mice to elucidate the role of angiotensin II through AT1a on the fibrosis of the obstructed kidney ( OBK ) .

This study established a UUO model in <protein>angiotensin type 1a receptor</protein> ( <protein>AT1a</protein> ) deficient ( mutant ) mice to elucidate the role of <protein>angiotensin II</protein> through <protein>AT1a</protein> on the fibrosis of the obstructed kidney ( OBK ) .


The relative volume of the tubulointerstitium was measured by an image analyzer ; deposition of collagen types III and IV and monocyte / macrophage infiltration were histologically examined using specific antibodies .

The relative volume of the tubulointerstitium was measured by an image analyzer ; deposition of <protein>collagen types III and IV</protein> and <cell_type>monocyte</cell_type> / <cell_type>macrophage</cell_type> infiltration were histologically examined using <protein>specific antibodies</protein> .


Also determined were the mRNA levels of transforming growth factor-beta by Northern blot analysis .

Also determined were the mRNA levels of <protein>transforming growth factor-beta</protein> by Northern blot analysis .


Nuclear factor-kappaB activity was assessed by gel shift assay .

<protein>Nuclear factor-kappaB</protein> activity was assessed by gel shift assay .


UUO in wild mice resulted in a marked expansion of relative volume of the tubulointerstitium , together with increased deposition of collagen types III and IV and number of infiltrated monocytes / macrophages in the interstitium , relative to sham-operated mice .

UUO in wild mice resulted in a marked expansion of relative volume of the tubulointerstitium , together with increased deposition of <protein>collagen types III and IV</protein> and number of infiltrated <cell_type>monocytes</cell_type> / <cell_type>macrophages</cell_type> in the interstitium , relative to sham-operated mice .


In comparison , these changes were significantly lower in mutant mice with UUO .


The mRNA level of transforming growth factor-beta was significantly higher in the OBK of wild mice with UUO compared with sham-operated mice .

The mRNA level of <protein>transforming growth factor-beta</protein> was significantly higher in the OBK of wild mice with UUO compared with sham-operated mice .


In contrast , the increase in mRNA level in the OBK of mutant mice was significantly less than in wild mice .


Finally , UUO resulted in activation of nuclear factor-kappaB in wild mice but was inhibited in the OBK of mutant mice .

Finally , UUO resulted in activation of <protein>nuclear factor-kappaB</protein> in wild mice but was inhibited in the OBK of mutant mice .


The results provide direct evidence that angiotensin II acting via the AT1a plays a pivotal role in the development of tubulointerstitial fibrosis in UUO .

The results provide direct evidence that <protein>angiotensin II</protein> acting via the <protein>AT1a</protein> plays a pivotal role in the development of tubulointerstitial fibrosis in UUO .


HLA-DQ tetramers identify epitope-specific T cells in peripheral blood of herpes simplex virus type 2-infected individuals : direct detection of immunodominant antigen-responsive cells .

<protein>HLA-DQ tetramers</protein> identify <cell_type>epitope-specific T cells</cell_type> in peripheral blood of herpes simplex virus type 2-infected individuals : direct detection of <cell_type>immunodominant antigen-responsive cells</cell_type> .


Ag-specific CD4+ T cells are present in peripheral blood in low frequency , where they undergo recruitment and expansion during immune responses and in the pathogenesis of numerous autoimmune diseases .

<cell_type>Ag-specific CD4+ T cells</cell_type> are present in peripheral blood in low frequency , where they undergo recruitment and expansion during immune responses and in the pathogenesis of numerous autoimmune diseases .


MHC tetramers , which constitute a labeled MHC-peptide ligand suitable for binding to the Ag-specific receptor on T cells , provide a novel approach for the detection and characterization of such rare cells .

<protein>MHC tetramers</protein> , which constitute a <protein>labeled MHC-peptide ligand</protein> suitable for binding to the <protein>Ag-specific receptor</protein> on <cell_type>T cells</cell_type> , provide a novel approach for the detection and characterization of such <cell_type>rare cells</cell_type> .


In this study , we utilized this technology to identify HLA DQ -restricted Ag-specific T cells in the peripheral blood of human subjects and to identify immunodominant epitopes associated with viral infection .

In this study , we utilized this technology to identify <protein>HLA DQ</protein> -restricted <cell_type>Ag-specific T cells</cell_type> in the peripheral blood of human subjects and to identify immunodominant epitopes associated with viral infection .


Peptides representing potential epitope regions of the VP16 protein from HSV-2 were loaded onto recombinant DQ0602 molecules to generate a panel of Ag-specific DQ0602 tetramers .

Peptides representing potential <protein>epitope regions</protein> of the <protein>VP16 protein</protein> from HSV-2 were loaded onto <protein>recombinant DQ0602 molecules</protein> to generate a panel of <protein>Ag-specific DQ0602 tetramers</protein> .


VP16 Ag -specific DQ-restricted T cells were identified and expanded from the peripheral blood of HSV-2-infected individuals , representing two predominant epitope specificities .

<protein>VP16 Ag</protein> -specific <cell_type>DQ-restricted T cells</cell_type> were identified and expanded from the peripheral blood of HSV-2-infected individuals , representing two predominant epitope specificities .


Although the VP16 369-380 peptide has a lower binding affinity for DQ0602 molecules than the VP16 33-52 peptide , T cells that recognized the VP16 369-380 peptide occurred at a much higher frequency than those that were specific for the VP16 33-52 peptide .

Although the VP16 369-380 peptide has a lower binding affinity for <protein>DQ0602 molecules</protein> than the VP16 33-52 peptide , <cell_type>T cells</cell_type> that recognized the VP16 369-380 peptide occurred at a much higher frequency than those that were specific for the VP16 33-52 peptide .


Down-regulation of BOB.1/OBF.1 and Oct2 in classical Hodgkin disease but not in lymphocyte predominant Hodgkin disease correlates with immunoglobulin transcription .

Down-regulation of <protein>BOB.1/OBF.1</protein> and <protein>Oct2</protein> in classical Hodgkin disease but not in lymphocyte predominant Hodgkin disease correlates with immunoglobulin transcription .


In contrast to the tumor cells ( L & H cells ) of lymphocyte predominant Hodgkin disease ( LPHD ) , Hodgkin and Reed-Sternberg ( HRS ) cells of classical Hodgkin disease ( cHD ) are unable to transcribe immunoglobulin , despite the presence of rearranged immunoglobulin genes .

In contrast to the <cell_type>tumor cells</cell_type> ( <cell_type>L & H cells</cell_type> ) of lymphocyte predominant Hodgkin disease ( LPHD ) , <cell_type>Hodgkin and Reed-Sternberg ( HRS ) cells</cell_type> of classical Hodgkin disease ( cHD ) are unable to transcribe <protein>immunoglobulin</protein> , despite the presence of <dna>rearranged immunoglobulin genes</dna> .


Although initial studies have suggested crippling immunoglobulin gene mutations to be the cause of absent immunoglobulin expression in cHD , recent work of our group has demonstrated an impaired activation of the immunoglobulin promoter as a superior mechanism .

Although initial studies have suggested crippling immunoglobulin gene mutations to be the cause of absent immunoglobulin expression in cHD , recent work of our group has demonstrated an impaired activation of the <dna>immunoglobulin promoter</dna> as a superior mechanism .


As immunoglobulin transcription is mainly regulated by the B-cell transcription factors Oct2 and BOB.1/OBF.1 , we analyzed 35 cases of LPHD , 32 cases of cHD , and 2 Hodgkin disease cell lines for the expression of these transcription factors and also in parallel for immunoglobulin expression .

As immunoglobulin transcription is mainly regulated by the <protein>B-cell transcription factors</protein> <protein>Oct2</protein> and <protein>BOB.1/OBF.1</protein> , we analyzed 35 cases of LPHD , 32 cases of cHD , and 2 <cell_line>Hodgkin disease cell lines</cell_line> for the expression of these <protein>transcription factors</protein> and also in parallel for immunoglobulin expression .


Our results demonstrate an absence of Oct2 and/or BOB.1/OBF.1 in cHD and a striking overexpression of Oct2 in LPHD .

Our results demonstrate an absence of <protein>Oct2</protein> and/or <protein>BOB.1/OBF.1</protein> in cHD and a striking overexpression of <protein>Oct2</protein> in LPHD .


Immunoglobulin expression was lacking in cHD but present in LPHD .


Furthermore , the reintroduction of BOB.1/OBF.1 and Oct2 into cultured HRS cells restored the activity of cotransduced immunoglobulin promoter constructs .

Furthermore , the reintroduction of <protein>BOB.1/OBF.1</protein> and <protein>Oct2</protein> into cultured <cell_type>HRS cells</cell_type> restored the activity of cotransduced <dna>immunoglobulin promoter constructs</dna> .


Our findings dismiss the concept that the different immunoglobulin expression in cHD and LPHD is due to disrupting mutations of immunoglobulin V genes in cHD but is most likely due to a down-regulation of Oct2 and/or BOB.1/OBF.1 .

Our findings dismiss the concept that the different immunoglobulin expression in cHD and LPHD is due to disrupting mutations of <dna>immunoglobulin V genes</dna> in cHD but is most likely due to a down-regulation of <protein>Oct2</protein> and/or <protein>BOB.1/OBF.1</protein> .


This study further revealed Oct2 as a new and valuable marker for the identification of L & H cells and their distinction from HRS cells .

This study further revealed <protein>Oct2</protein> as a new and valuable marker for the identification of <cell_type>L & H cells</cell_type> and their distinction from <cell_type>HRS cells</cell_type> .


The impairment of immunoglobulin transcription with a down-regulated synthesis of Oct2 and BOB.1/OBF.1 is the first established general recurrent defect found in HRS cells .

The impairment of immunoglobulin transcription with a down-regulated synthesis of <protein>Oct2</protein> and <protein>BOB.1/OBF.1</protein> is the first established general recurrent defect found in <cell_type>HRS cells</cell_type> .


Constitutive expression of NF-kappa B is a characteristic feature of mycosis fungoides : implications for apoptosis resistance and pathogenesis .

Constitutive expression of <protein>NF-kappa B</protein> is a characteristic feature of mycosis fungoides : implications for apoptosis resistance and pathogenesis .


The NF-kappa B family of transcription factors is an important regulator of genes expressed during inflammatory responses , immunoglobulin ( Ig ) class switching , cellular differentiation , and apoptosis .

The <protein>NF-kappa B family of transcription factors</protein> is an important regulator of genes expressed during inflammatory responses , immunoglobulin ( Ig ) class switching , cellular differentiation , and apoptosis .


Recently , members of the NF-kappaB family , including p65 ( Rel A ) , have been implicated in promoting survival of various hematopoeitic neoplasms , including T cell malignancies such as adult T cell leukemia-lymphoma .

Recently , members of the <protein>NF-kappaB family</protein> , including <protein>p65 ( Rel A )</protein> , have been implicated in promoting survival of various hematopoeitic neoplasms , including T cell malignancies such as adult T cell leukemia-lymphoma .


We investigated the expression of active NF-kappa B p65 ( Rel A ) in cases of mycosis fungoides ( MF ) and the effect of chemical inhibitors of NF-kappa B on apoptosis in cutaneous T cell lymphoma ( CTCL ) cell lines .

We investigated the expression of <protein>active NF-kappa B p65 ( Rel A )</protein> in cases of mycosis fungoides ( MF ) and the effect of chemical inhibitors of <protein>NF-kappa B</protein> on apoptosis in <cell_line>cutaneous T cell lymphoma ( CTCL ) cell lines</cell_line> .


Paraffin-embedded tissues from 23 cutaneous lesions and a single lymph node biopsy from patients diagnosed with MF were evaluated for p65 ( Rel A ) expression by using a monoclonal mouse antibody that detects the activated form of p65 ( Rel A ) .

Paraffin-embedded tissues from 23 cutaneous lesions and a single lymph node biopsy from patients diagnosed with MF were evaluated for p65 ( Rel A ) expression by using a <protein>monoclonal mouse antibody</protein> that detects the activated form of <protein>p65 ( Rel A )</protein> .


Apoptosis after treatment with the NF-kappa B inhibitors gliotoxin , MG132 , BAY 11-7082 , and BAY 11-7085 was quantitatively measured in the CTCL cell lines HuT-78 and HH by propidium iodide ( PI ) /cell cycle analysis for detection of a hypodiploid ( sub-G ( 0 ) ) population and by determination of increased Annexin V/7-amino-actinomycin D ( 7-AAD ) expression .

Apoptosis after treatment with the NF-kappa B inhibitors gliotoxin , MG132 , BAY 11-7082 , and BAY 11-7085 was quantitatively measured in the <cell_line>CTCL cell lines HuT-78 and HH</cell_line> by propidium iodide ( PI ) /cell cycle analysis for detection of a hypodiploid ( sub-G ( 0 ) ) population and by determination of increased Annexin V/7-amino-actinomycin D ( 7-AAD ) expression .


Nuclear extracts from CTCL cells before and after chemical inhibition were analyzed for NF-kappa B nuclear DNA-binding activity by electrophoretic mobility shift assay ( EMSA ) with quantitative densitometry .

Nuclear extracts from <cell_type>CTCL cells</cell_type> before and after chemical inhibition were analyzed for NF-kappa B nuclear DNA-binding activity by electrophoretic mobility shift assay ( EMSA ) with quantitative densitometry .


Nuclear expression of p65 ( Rel A ) before and after treatment with the various inhibitory compounds was measured by immunofluorescence staining in each CTCL cell line .

Nuclear expression of <protein>p65 ( Rel A )</protein> before and after treatment with the various inhibitory compounds was measured by immunofluorescence staining in each <cell_line>CTCL cell line</cell_line> .


Neoplastic T lymphocytes from 22 of 24 cases of MF showed strong nuclear and cytoplasmic expression of active p65 ( Rel A ) .

<cell_type>Neoplastic T lymphocytes</cell_type> from 22 of 24 cases of MF showed strong nuclear and cytoplasmic expression of <protein>active p65 ( Rel A )</protein> .


Compared with untreated control cells , a marked increase in apoptosis , a significant decrease in NF-kappa B DNA-binding activity , and a marked decrease in nuclear p65 ( Rel A ) expression were seen in cells from both CTCL cell lines after chemical NF-kappa B inhibition .

Compared with untreated <cell_type>control cells</cell_type> , a marked increase in apoptosis , a significant decrease in NF-kappa B DNA-binding activity , and a marked decrease in nuclear p65 ( Rel A ) expression were seen in cells from both <cell_line>CTCL cell lines</cell_line> after chemical NF-kappa B inhibition .


These data show that the active form of NF-kappa B p65 ( Rel A ) is commonly expressed in neoplastic T lymphocytes in patients with MF .

These data show that the active form of <protein>NF-kappa B p65 ( Rel A )</protein> is commonly expressed in <cell_type>neoplastic T lymphocytes</cell_type> in patients with MF .


In CTCL cell lines , the significant decrease in nuclear NF-kappa B expression and the marked increase in spontaneous apoptosis caused by chemical NF-kappa B inhibition suggest a critical role for NF-kappa B in the pathogenesis and tumor cell maintenance of CTCLs .

In <cell_line>CTCL cell lines</cell_line> , the significant decrease in nuclear NF-kappa B expression and the marked increase in spontaneous apoptosis caused by chemical NF-kappa B inhibition suggest a critical role for <protein>NF-kappa B</protein> in the pathogenesis and tumor cell maintenance of CTCLs .


HUM PATHOL 31 : 1482-1490 .


Human T-cell leukemia virus type 1 tax protein activates transcription through AP-1 site by inducing DNA binding activity in T cells .

Human T-cell leukemia virus type 1 <protein>tax protein</protein> activates transcription through <dna>AP-1 site</dna> by inducing DNA binding activity in <cell_type>T cells</cell_type> .


Human T-cell leukemia virus type 1 ( HTLV-1 ) Tax protein induces the expression of various family members of the transcription factor AP-1 , such as c-Jun , JunD , c-Fos , and Fra-1 , at the level of RNA expression in T cells .

Human T-cell leukemia virus type 1 ( HTLV-1 ) <protein>Tax protein</protein> induces the expression of various family members of the <protein>transcription factor AP-1</protein> , such as <protein>c-Jun</protein> , <protein>JunD</protein> , <protein>c-Fos</protein> , and <protein>Fra-1</protein> , at the level of RNA expression in <cell_type>T cells</cell_type> .


We examined the activity of Tax in transcription through AP-1-binding sites ( AP-1 site ) in T cells .

We examined the activity of <protein>Tax</protein> in transcription through <dna>AP-1-binding sites</dna> ( <dna>AP-1 site</dna> ) in <cell_type>T cells</cell_type> .


Transient transfection studies showed that Tax activated the expression of a luciferase gene regulated by two copies of an AP-1 site in the human Jurkat T-cell line .

Transient transfection studies showed that <protein>Tax</protein> activated the expression of a <dna>luciferase gene</dna> regulated by two copies of an <dna>AP-1 site</dna> in the <cell_line>human Jurkat T-cell line</cell_line> .


Tax activates the expression of viral and cellular genes through two different enhancers : a cAMP-responsive ( CRE ) -like element and a kappaB element .

<protein>Tax</protein> activates the expression of <dna>viral and cellular genes</dna> through two different enhancers : a <dna>cAMP-responsive ( CRE ) -like element</dna> and a <dna>kappaB element</dna> .


Two Tax mutants differentially activated expression of these two elements .

Two <protein>Tax mutants</protein> differentially activated expression of these two elements .


Tax703 preferentially activated the kappaB element but not the CRE-like one , whereas TaxM22 showed the reverse .

<protein>Tax703</protein> preferentially activated the <dna>kappaB element</dna> but not the CRE-like one , whereas <protein>TaxM22</protein> showed the reverse .


In addition , Tax703 and Tax , but not TaxM22 , converted cell growth of a mouse T-cell line from being interleukin ( IL ) -2-dependent to being IL-2-independent .

In addition , <protein>Tax703</protein> and <protein>Tax</protein> , but not <protein>TaxM22</protein> , converted cell growth of a <cell_line>mouse T-cell line</cell_line> from being interleukin ( IL ) -2-dependent to being IL-2-independent .


Unlike the wild-type Tax , Tax703 and TaxM22 only weakly activated the AP-1 site in the T-cell line .

Unlike the <protein>wild-type Tax</protein> , <protein>Tax703</protein> and <protein>TaxM22</protein> only weakly activated the <dna>AP-1 site</dna> in the <cell_line>T-cell line</cell_line> .


Thus , Tax seems to activate the AP-1 site via mechanisms distinct from those of kappaB or CRE-like elements , and the activation of the AP-1 site is dispensable for IL-2-independent growth of CTLL-2 .

Thus , <protein>Tax</protein> seems to activate the <dna>AP-1 site</dna> via mechanisms distinct from those of <dna>kappaB or CRE-like elements</dna> , and the activation of the <dna>AP-1 site</dna> is dispensable for IL-2-independent growth of CTLL-2 .


Electrophoretic mobility shift assays showed that Tax induced strong binding activity to an AP-1 site in CTLL-2 , whereas Tax703 did not , indicating that the induction of binding activity to the AP-1 site is essential for the transcriptional activation by Tax .

Electrophoretic mobility shift assays showed that <protein>Tax</protein> induced strong binding activity to an <dna>AP-1 site</dna> in CTLL-2 , whereas <protein>Tax703</protein> did not , indicating that the induction of binding activity to the <dna>AP-1 site</dna> is essential for the transcriptional activation by <protein>Tax</protein> .


The binding complex induced by Tax in CTLL-2 contained JunD and Fra-2 .

The <protein>binding complex</protein> induced by <protein>Tax</protein> in CTLL-2 contained <protein>JunD</protein> and <protein>Fra-2</protein> .


Other AP-1 proteins were undetectable .

Other <protein>AP-1 proteins</protein> were undetectable .


Activation of transcription through the AP-1 site in Jurkat cells by JunD and/or Fra-2 was weak .

Activation of transcription through the <dna>AP-1 site</dna> in <cell_line>Jurkat cells</cell_line> by <protein>JunD</protein> and/or <protein>Fra-2</protein> was weak .


c-Jun , JunB , and c-Fos activation was greater , although the level was still less than that with Tax .

<protein>c-Jun</protein> , <protein>JunB</protein> , and <protein>c-Fos</protein> activation was greater , although the level was still less than that with <protein>Tax</protein> .


Thus , the induction of AP-1 mRNA by Tax may not be sufficient for a complete activation of AP-1 site by Tax .

Thus , the induction of <rna>AP-1 mRNA</rna> by <protein>Tax</protein> may not be sufficient for a complete activation of <dna>AP-1 site</dna> by <protein>Tax</protein> .


Our results suggest that Tax activates the transcription of cellular genes with AP-1 sites by inducing the DNA-binding activity of AP-1 proteins in T cells , a mechanism distinct from those of CRE-like and kappaB elements .

Our results suggest that <protein>Tax</protein> activates the transcription of <dna>cellular genes</dna> with <dna>AP-1 sites</dna> by inducing the DNA-binding activity of <protein>AP-1 proteins</protein> in <cell_type>T cells</cell_type> , a mechanism distinct from those of <dna>CRE-like and kappaB elements</dna> .


Copyright 2001 Academic Press .


Strict control of human immunodeficiency virus type 1 replication by a genetic switch : Tet for Tat .


Live-attenuated human immunodeficiency virus type 1 ( HIV-1 ) variants have shown great promise as AIDS vaccines , but continued replication can lead to the selection of faster-replicating variants that are pathogenic .


We therefore designed HIV-1 genomes that replicate exclusively upon addition of the nontoxic effector doxycycline ( dox ) .

We therefore designed <dna>HIV-1 genomes</dna> that replicate exclusively upon addition of the nontoxic effector doxycycline ( dox ) .


This was achieved by replacement of the viral TAR-Tat system for transcriptional activation by the Escherichia coli-derived Tet system for inducible gene expression .


These designer `` HIV-rtTA '' viruses replicate in a strictly dox-dependent manner both in a T-cell line and in primary blood cells , and the rate of replication can be fine-tuned by simple variation of the dox concentration .

These designer `` HIV-rtTA '' viruses replicate in a strictly dox-dependent manner both in a <cell_line>T-cell line</cell_line> and in <cell_type>primary blood cells</cell_type> , and the rate of replication can be fine-tuned by simple variation of the dox concentration .


These HIV-rtTA viruses provide a tool to perform genetics , e.g. , selection and optimization experiments , with the E. coli-derived Tet reagents in a eukaryotic background .


Furthermore , such viruses may represent improved vaccine candidates because their replication can be turned on and off at will .


Synovial fluid induced nuclear factor-kappaB DNA binding in a monocytic cell line .

Synovial fluid induced <protein>nuclear factor-kappaB</protein> DNA binding in a <cell_line>monocytic cell line</cell_line> .


OBJECTIVE : To determine the effects of synovial fluids ( SF ) on DNA binding activity of transcription factor nuclear factor-kappaB ( NF-kappaB ) in the Mono Mac 6 monocytic/macrophage cell line as a model for the interaction between SF and synovial tissue macrophages in arthritis .

OBJECTIVE : To determine the effects of synovial fluids ( SF ) on DNA binding activity of <protein>transcription factor</protein> <protein>nuclear factor-kappaB</protein> ( <protein>NF-kappaB</protein> ) in the <cell_line>Mono Mac 6 monocytic/macrophage cell line</cell_line> as a model for the interaction between SF and <cell_type>synovial tissue macrophages</cell_type> in arthritis .


METHODS : Mono Mac 6 cells were incubated with SF from the knee joints of human subjects with rheumatoid arthritis ( RA ) , undifferentiated seronegative oligoarthritis , and osteoarthritis ( OA ) .

METHODS : <cell_line>Mono Mac 6 cells</cell_line> were incubated with SF from the knee joints of human subjects with rheumatoid arthritis ( RA ) , undifferentiated seronegative oligoarthritis , and osteoarthritis ( OA ) .


Nuclear extracts prepared from the Mono Mac 6 cells and RA synovial tissue were analyzed by electrophoretic mobility shift analysis ( EMSA ) for NF-kappaB DNA binding proteins .

Nuclear extracts prepared from the <cell_line>Mono Mac 6 cells</cell_line> and RA synovial tissue were analyzed by electrophoretic mobility shift analysis ( EMSA ) for <protein>NF-kappaB DNA binding proteins</protein> .


RESULTS : Induction of NF-kappaB DNA binding by the p65 ( RelA ) /p50 heterodimer was observed in response to incubation of Mono Mac 6 cells with SF ( 20 % in culture medium ) from 5 of 8 subjects with RA , 4 of 5 with OA , and none of 3 with undifferentiated seronegative oligoarthritis .

RESULTS : Induction of <protein>NF-kappaB</protein> DNA binding by the <protein>p65 ( RelA ) /p50 heterodimer</protein> was observed in response to incubation of <cell_line>Mono Mac 6 cells</cell_line> with SF ( 20 % in culture medium ) from 5 of 8 subjects with RA , 4 of 5 with OA , and none of 3 with undifferentiated seronegative oligoarthritis .


Incubation of SF with neutralizing antibodies against tumor necrosis factor-alpha ( TNF-alpha ) , but not antibodies against interleukin 6 ( IL-6 ) , significantly reduced the induction of p65/p50 binding activity in SF from subjects with RA and OA .

Incubation of SF with <protein>neutralizing antibodies</protein> against <protein>tumor necrosis factor-alpha ( TNF-alpha )</protein> , but not <protein>antibodies</protein> against <protein>interleukin 6 ( IL-6 )</protein> , significantly reduced the induction of p65/p50 binding activity in SF from subjects with RA and OA .


Unexpectedly , a slowly migrating SF inducible NF-kappaB -binding complex was observed in EMSA of Mono Mac 6 cells after incubation with SF from 5 of 8 RA and 2 of 5 OA subjects .

Unexpectedly , a slowly migrating SF inducible <protein>NF-kappaB</protein> -binding complex was observed in EMSA of <cell_line>Mono Mac 6 cells</cell_line> after incubation with SF from 5 of 8 RA and 2 of 5 OA subjects .


The induction of this complex by SF was not affected by neutralization of TNF-alpha or IL-6 in SF , and the complex was not inducible by TNF-alpha , IL-1beta , TNF-alpha / IL-1beta , IL-6 , platelet derived growth factor , lipopolysaccharide , or tetradecanoyl phorbol acetate .

The induction of this complex by SF was not affected by neutralization of <protein>TNF-alpha</protein> or <protein>IL-6</protein> in SF , and the complex was not inducible by <protein>TNF-alpha</protein> , IL-1beta , <protein>TNF-alpha</protein> / <protein>IL-1beta</protein> , <protein>IL-6</protein> , <protein>platelet derived growth factor</protein> , lipopolysaccharide , or tetradecanoyl phorbol acetate .


The slowly migrating complex could not be supershifted with antibodies against NF-kappaB , Jun , or the transcriptional coactivators p300 or CBP .

The slowly <protein>migrating complex</protein> could not be supershifted with <protein>antibodies</protein> against <protein>NF-kappaB</protein> , <protein>Jun</protein> , or the transcriptional coactivators <protein>p300</protein> or <protein>CBP</protein> .


A NF-kappaB-binding complex with similar slow mobility was observed in nuclear extracts prepared from fresh human RA synovial tissue .

A <protein>NF-kappaB-binding complex</protein> with similar slow mobility was observed in nuclear extracts prepared from fresh human RA synovial tissue .


CONCLUSION : Biological activity of TNF-alpha in SF from RA and OA subjects is capable of inducing p65/p50 NF-kappaB DNA binding activity in macrophages .

CONCLUSION : Biological activity of <protein>TNF-alpha</protein> in SF from RA and OA subjects is capable of inducing <protein>p65/p50 NF-kappaB</protein> DNA binding activity in <cell_type>macrophages</cell_type> .


A property of SF that is independent of TNF-alpha and other cytokines is responsible for the induction of a novel slowly migrating NF-kappaB-binding complex .

A property of SF that is independent of <protein>TNF-alpha</protein> and other <protein>cytokines</protein> is responsible for the induction of a novel slowly migrating <protein>NF-kappaB-binding complex</protein> .


Soluble mediators in SF of subjects with RA and OA can therefore modulate binding of nuclear proteins to the NF-kappaB binding site in macrophages and may play a role in inflammatory gene expression in arthritis .

Soluble mediators in SF of subjects with RA and OA can therefore modulate binding of <protein>nuclear proteins</protein> to the <protein>NF-kappaB binding site</protein> in <cell_type>macrophages</cell_type> and may play a role in inflammatory gene expression in arthritis .


Peroxisome proliferator activator receptor-gamma agonists and 15-deoxy-Delta ( 12 , 14 ) ( 12 , 14 ) -PGJ ( 2 ) induce apoptosis in normal and malignant B-lineage cells .

<protein>Peroxisome proliferator activator receptor-gamma</protein> agonists and 15-deoxy-Delta ( 12 , 14 ) ( 12 , 14 ) -PGJ ( 2 ) induce apoptosis in normal and malignant <cell_type>B-lineage cells</cell_type> .


The research described herein evaluates the expression and functional significance of peroxisome proliferator activator receptor-gamma ( PPAR-gamma ) on B-lineage cells .

The research described herein evaluates the expression and functional significance of <protein>peroxisome proliferator activator receptor-gamma ( PPAR-gamma )</protein> on <cell_type>B-lineage cells</cell_type> .


Normal mouse B cells and a variety of B lymphoma cells reflective of stages of B cell differentiation ( e.g. , 70Z/3 , CH31 , WEHI-231 , CH12 , and J558 ) express PPAR-gamma mRNA and , by Western blot analysis , the 67-kDa PPAR-gamma protein .

<cell_type>Normal mouse B cells</cell_type> and a variety of <cell_type>B lymphoma cells</cell_type> reflective of stages of B cell differentiation ( e.g. , 70Z/3 , CH31 , WEHI-231 , CH12 , and J558 ) express <rna>PPAR-gamma mRNA</rna> and , by Western blot analysis , the <protein>67-kDa PPAR-gamma protein</protein> .


15-Deoxy-Delta ( 12 , 14 ) -PGJ ( 2 ) ( 15d-PGJ ( 2 ) ) , a PPAR-gamma agonist , has a dose-dependent antiproliferative and cytotoxic effect on normal and malignant B cells as shown by [ ( 3 ) H ] thymidine and 3- [ 4 , 5-dimethylthiazol-2-yl ] -2 , 5-diphenyltetrazolium bromide assays .

15-Deoxy-Delta ( 12 , 14 ) -PGJ ( 2 ) ( 15d-PGJ ( 2 ) ) , a PPAR-gamma agonist , has a dose-dependent antiproliferative and cytotoxic effect on <cell_type>normal and malignant B cells</cell_type> as shown by [ ( 3 ) H ] thymidine and 3- [ 4 , 5-dimethylthiazol-2-yl ] -2 , 5-diphenyltetrazolium bromide assays .


Only PPAR-gamma agonists ( thiazolidinediones ) , and not PPAR-alpha agonists , mimicked the effect of 15d-PGJ ( 2 ) on B-lineage cells , indicating that the mechanism by which 15d-PGJ ( 2 ) negatively affects B-lineage cells involves in part PPAR-gamma .

Only PPAR-gamma agonists ( thiazolidinediones ) , and not PPAR-alpha agonists , mimicked the effect of 15d-PGJ ( 2 ) on <cell_type>B-lineage cells</cell_type> , indicating that the mechanism by which 15d-PGJ ( 2 ) negatively affects <cell_type>B-lineage cells</cell_type> involves in part <protein>PPAR-gamma</protein> .


The mechanism by which PPAR-gamma agonists induce cytotoxicity is via apoptosis , as shown by annexin V staining and as confirmed by DNA fragmentation detected using the TUNEL assay .


Interestingly , addition of PGF ( 2alpha ) , which was not known to affect lymphocytes , dramatically attenuated the deleterious effects of PPAR-gamma agonists on B lymphomas .

Interestingly , addition of PGF ( 2alpha ) , which was not known to affect <cell_type>lymphocytes</cell_type> , dramatically attenuated the deleterious effects of PPAR-gamma agonists on <cell_type>B lymphomas</cell_type> .


Surprisingly , 15d-PGJ ( 2 ) induced a massive increase in nuclear mitogen-activated protein kinase activation , and pretreatment with PGF ( 2alpha ) blunted the mitogen-activated protein kinase activation .


This is the first study evaluating PPAR-gamma expression and its significance on B lymphocytes .

This is the first study evaluating PPAR-gamma expression and its significance on <cell_type>B lymphocytes</cell_type> .


PPAR-gamma agonists may serve as a counterbalance to the stimulating effects of other PGs , namely PGE ( 2 ) , which promotes B cell differentiation .


Finally , the use of PGs , such as 15d-PGJ ( 2 ) , and synthetic PPAR-gamma agonists to induce apoptosis in B-lineage cells may lead to the development of novel therapies for fatal B lymphomas

Finally , the use of PGs , such as 15d-PGJ ( 2 ) , and synthetic PPAR-gamma agonists to induce apoptosis in <cell_type>B-lineage cells</cell_type> may lead to the development of novel therapies for <cell_type>fatal B lymphomas</cell_type>


Inhibition of STAT3 signaling leads to apoptosis of leukemic large granular lymphocytes and decreased Mcl-1 expression .

Inhibition of STAT3 signaling leads to apoptosis of <cell_type>leukemic large granular lymphocytes</cell_type> and decreased Mcl-1 expression .


Large granular lymphocyte ( LGL ) leukemia is characterized by the expansion of antigen-activated cytotoxic T lymphocytes .

Large granular lymphocyte ( LGL ) leukemia is characterized by the expansion of <cell_type>antigen-activated cytotoxic T lymphocytes</cell_type> .


These leukemic cells are resistant to Fas-mediated apoptosis despite expressing high levels of Fas .

These <cell_type>leukemic cells</cell_type> are resistant to Fas-mediated apoptosis despite expressing high levels of <protein>Fas</protein> .


We found that leukemic LGL from 19 patients displayed high levels of activated STAT3 .

We found that <cell_type>leukemic LGL</cell_type> from 19 patients displayed high levels of <protein>activated STAT3</protein> .


Treatment of leukemic LGL with the JAK-selective tyrosine kinase inhibitor AG-490 induced apoptosis with a corresponding decrease in STAT -DNA binding activity .

Treatment of <cell_type>leukemic LGL</cell_type> with the JAK-selective tyrosine kinase inhibitor AG-490 induced apoptosis with a corresponding decrease in <protein>STAT</protein> -DNA binding activity .


Moreover , using an antisense oligonucleotide approach to diminish STAT3 expression , we found that Fas sensitivity was restored in leukemic LGL .

Moreover , using an antisense oligonucleotide approach to diminish STAT3 expression , we found that Fas sensitivity was restored in <cell_type>leukemic LGL</cell_type> .


AG-490-induced apoptosis in leukemic LGL was independent of Bcl-xL or Bcl-2 expression .

AG-490-induced apoptosis in <cell_type>leukemic LGL</cell_type> was independent of Bcl-xL or Bcl-2 expression .


However , we found that the Bcl-2-family protein Mcl-1 was significantly reduced by AG-490 treatment .

However , we found that the <protein>Bcl-2-family protein Mcl-1</protein> was significantly reduced by AG-490 treatment .


Activated STAT3 was shown to bind an SIE-related element in the murine mcl-1 promoter .

<protein>Activated STAT3</protein> was shown to bind an <dna>SIE-related element</dna> in the <dna>murine mcl-1 promoter</dna> .


Using a luciferase reporter assay , we demonstrated that v-src overexpression in NIH3T3 induced STAT3 -dependent transcriptional activity from the mcl-1 promoter and increased endogenous Mcl-1 protein levels .

Using a luciferase reporter assay , we demonstrated that v-src overexpression in NIH3T3 induced <protein>STAT3</protein> -dependent transcriptional activity from the <dna>mcl-1 promoter</dna> and increased endogenous Mcl-1 protein levels .


We conclude that STAT3 activation contributed to accumulation of the leukemic LGL clones .

We conclude that <protein>STAT3</protein> activation contributed to accumulation of the <cell_type>leukemic LGL clones</cell_type> .


These findings suggest that investigation should focus on novel strategies targeting STAT3 in the treatment of LGL leukemia .

These findings suggest that investigation should focus on novel strategies targeting <protein>STAT3</protein> in the treatment of LGL leukemia .


Decreased immediate inflammatory gene induction in activating transcription factor-2 mutant mice .

Decreased immediate <dna>inflammatory gene</dna> induction in <protein>activating transcription factor-2</protein> mutant mice .


Transcription factor activating transcription factor ( ATF ) -2 is activated by inflammatory signals transduced by the JNK and p38 MAP kinase pathways .

<protein>Transcription factor activating transcription factor ( ATF ) -2</protein> is activated by inflammatory signals transduced by the JNK and p38 MAP kinase pathways .


To better define the role of ATF-2 in inflammation , adult mice expressing small amounts of a mutant ATF-2 protein were challenged with lipopolysaccharide ( LPS ) , anti-CD3 antibody or virus .

To better define the role of <protein>ATF-2</protein> in inflammation , adult mice expressing small amounts of a <protein>mutant ATF-2 protein</protein> were challenged with lipopolysaccharide ( LPS ) , <protein>anti-CD3 antibody</protein> or virus .


Within 3 h of challenge by LPS , ATF-2 mutant mice had decreased induction of the adhesion molecules E-selectin , P-selectin and VCAM-1 as well as the cytokines tumor necrosis factor-alpha , IL-1beta and IL-6 compared with control mice .

Within 3 h of challenge by LPS , ATF-2 mutant mice had decreased induction of the <protein>adhesion molecules</protein> <protein>E-selectin</protein> , <protein>P-selectin</protein> and <protein>VCAM-1</protein> as well as the <protein>cytokines</protein> <protein>tumor necrosis factor-alpha</protein> , <protein>IL-1beta</protein> and <protein>IL-6</protein> compared with control mice .


Stimulation of T lymphocytes by anti-CD3 antibody also showed less induction of IL-1 and IL-6 in ATF-2 mutant tissues .

Stimulation of <cell_type>T lymphocytes</cell_type> by <protein>anti-CD3 antibody</protein> also showed less induction of <protein>IL-1</protein> and <protein>IL-6</protein> in ATF-2 mutant tissues .


ATF-2 mutant thymocytes treated with anti-CD3 antibody in vitro demonstrated reduced induction of c-Jun , JunB , JunD and Fra-2 .

<cell_type>ATF-2 mutant thymocytes</cell_type> treated with <protein>anti-CD3 antibody</protein> in vitro demonstrated reduced induction of <protein>c-Jun</protein> , <protein>JunB</protein> , <protein>JunD</protein> and <protein>Fra-2</protein> .


However , similar to what was observed after p38 kinase inhibition in normal mice , relative ATF-2 deficiency did not prevent the development of a mononuclear cell infiltrate in the week following an inflammatory stimulus .

However , similar to what was observed after p38 kinase inhibition in normal mice , relative ATF-2 deficiency did not prevent the development of a <cell_type>mononuclear cell</cell_type> infiltrate in the week following an inflammatory stimulus .


ATF-2 mutant mice proved more susceptible to death than control mice from LPS plus D-galactosamine injection or Coxsackievirus B3 infection and had a higher incidence of mononuclear pulmonary infiltrates after exposure to Herpes simplex virus-1 .


ATF-2 is essential for maximal immediate induction of adhesion molecules and cytokine genes , but at later time points may even protect against overactive immune responses .

<protein>ATF-2</protein> is essential for maximal immediate induction of <protein>adhesion molecules</protein> and <dna>cytokine genes</dna> , but at later time points may even protect against overactive immune responses .


LIGHT , a TNF-like molecule , costimulates T cell proliferation and is required for dendritic cell -mediated allogeneic T cell response .

<protein>LIGHT</protein> , a <protein>TNF-like molecule</protein> , costimulates T cell proliferation and is required for <cell_type>dendritic cell</cell_type> -mediated allogeneic T cell response .


LIGHT is a recently identified member of the TNF superfamily and its receptors , herpesvirus entry mediator and lymphotoxin beta receptor , are found in T cells and stromal cells .

<protein>LIGHT</protein> is a recently identified member of the <protein>TNF superfamily</protein> and its receptors , <protein>herpesvirus entry mediator</protein> and <protein>lymphotoxin beta receptor</protein> , are found in <cell_type>T cells</cell_type> and <cell_type>stromal cells</cell_type> .


In this study , we demonstrate that LIGHT is selectively expressed on immature dendritic cells ( DCs ) generated from human PBMCs .

In this study , we demonstrate that <protein>LIGHT</protein> is selectively expressed on <cell_type>immature dendritic cells</cell_type> ( <cell_type>DCs</cell_type> ) generated from <cell_type>human PBMCs</cell_type> .


In contrast , LIGHT is not detectable in DCs either freshly isolated from PBMCs or rendered mature in vitro by LPS treatment .

In contrast , <protein>LIGHT</protein> is not detectable in <cell_type>DCs</cell_type> either freshly isolated from <cell_type>PBMCs</cell_type> or rendered mature in vitro by LPS treatment .


Blockade of LIGHT by its soluble receptors , lymphotoxin beta receptor-Ig or HVEM-Ig , inhibits the induction of DC -mediated primary allogeneic T cell response .

Blockade of <protein>LIGHT</protein> by its <protein>soluble receptors</protein> , <protein>lymphotoxin beta receptor-Ig</protein> or <protein>HVEM-Ig</protein> , inhibits the induction of <cell_type>DC</cell_type> -mediated primary allogeneic T cell response .


Furthermore , engagement of LIGHT costimulates human T cell proliferation , amplifies the NF-kappaB signaling pathway , and preferentially induces the production of IFN-gamma , but not IL-4 , in the presence of an antigenic signal .

Furthermore , engagement of <protein>LIGHT</protein> costimulates human T cell proliferation , amplifies the NF-kappaB signaling pathway , and preferentially induces the production of <protein>IFN-gamma</protein> , but not <protein>IL-4</protein> , in the presence of an antigenic signal .


Our results suggest that LIGHT is a costimulatory molecule involved in DC -mediated cellular immune responses .

Our results suggest that <protein>LIGHT</protein> is a <protein>costimulatory molecule</protein> involved in <cell_type>DC</cell_type> -mediated cellular immune responses .


Suppression of HIV type 1 replication by a dominant-negative Ets-1 mutant .

Suppression of HIV type 1 replication by a <protein>dominant-negative Ets-1 mutant</protein> .


Activity of the distal region of the human immunodeficiency virus ( HIV-1 ) long terminal repeat ( LTR ) , which contains binding sites for the Ets-1 and USF-1 proteins , is integral for HIV-1 replication .

Activity of the <dna>distal region</dna> of the human immunodeficiency virus ( HIV-1 ) <dna>long terminal repeat</dna> ( <dna>LTR</dna> ) , which contains <dna>binding sites</dna> for the <protein>Ets-1 and USF-1 proteins</protein> , is integral for HIV-1 replication .


The Ets-1 and USF-1 proteins play a critical role in the activity of the HIV-1 LTR distal enhancer region , as indicated by the potent dominant negative effect of a mutant Ets-1 lacking trans-activation domains on the transcriptional activity of the LTR .

The <protein>Ets-1 and USF-1 proteins</protein> play a critical role in the activity of the <dna>HIV-1 LTR distal enhancer region</dna> , as indicated by the potent dominant negative effect of a <dna>mutant Ets-1 lacking trans-activation domains</dna> on the transcriptional activity of the <dna>LTR</dna> .


To determine the biological relevance of the Ets-1 and USF-1 proteins in HIV-1 replication , we examined the effect of expression of the dominant-negative mutant of Ets-1 ( dnEts-1 ) on HIV-1 infection of T cells .

To determine the biological relevance of the <protein>Ets-1 and USF-1 proteins</protein> in HIV-1 replication , we examined the effect of expression of the dominant-negative mutant of <dna>Ets-1</dna> ( <dna>dnEts-1</dna> ) on HIV-1 infection of <cell_type>T cells</cell_type> .


We demonstrated that expression of dnEts markedly suppressed HIV-1 infection of a T cell line .

We demonstrated that expression of <dna>dnEts</dna> markedly suppressed HIV-1 infection of a <cell_line>T cell line</cell_line> .


This finding indicates that formation of a transcriptionaly active USF-1/Ets-1 complex is important in the productive infection of cells by HIV-1 , and suggests that inhibition of the interaction between USF-1 and Ets-1 with the HIV-1 LTR may provide a new target for anti-HIV-1 gene therapy .

This finding indicates that formation of a <protein>transcriptionaly active USF-1/Ets-1 complex</protein> is important in the productive infection of cells by HIV-1 , and suggests that inhibition of the interaction between <protein>USF-1</protein> and <protein>Ets-1</protein> with the <dna>HIV-1 LTR</dna> may provide a new target for anti-HIV-1 gene therapy .


Human eosinophils constitutively express nuclear factor of activated T cells p and c .

<cell_type>Human eosinophils</cell_type> constitutively express <protein>nuclear factor of activated T cells p and c</protein> .


BACKGROUND : Eosinophils are now known to produce a variety of proinflammatory cytokines , although the molecular factors that regulate their production are poorly understood .

BACKGROUND : <cell_type>Eosinophils</cell_type> are now known to produce a variety of <protein>proinflammatory cytokines</protein> , although the molecular factors that regulate their production are poorly understood .


The expression of almost all of the cytokines produced by eosinophils , including the proallergic cytokine IL-4 , is now known to be regulated at the level of transcription by members of the nuclear factor of activated T cells ( NFAT ) family of transcription factors .

The expression of almost all of the <protein>cytokines</protein> produced by <cell_type>eosinophils</cell_type> , including the <protein>proallergic cytokine IL-4</protein> , is now known to be regulated at the level of transcription by members of the <protein>nuclear factor of activated T cells</protein> ( <protein>NFAT</protein> ) family of transcription factors .


OBJECTIVE : We sought to characterize the expression of different NFAT proteins in resting and activated eosinophils .

OBJECTIVE : We sought to characterize the expression of different <protein>NFAT proteins</protein> in <cell_type>resting and activated eosinophils</cell_type> .


METHODS : Nuclear and whole cell extracts were obtained from both peripheral blood eosinophils and those obtained from bronchoalveolar lavage fluid of asthmatic subjects after endobronchial allergen challenge .

METHODS : Nuclear and whole cell extracts were obtained from both <cell_type>peripheral blood eosinophils</cell_type> and those obtained from bronchoalveolar lavage fluid of asthmatic subjects after endobronchial allergen challenge .


NFAT expression was determined by using immunoprecipitation and Western blot analysis , DNA-binding assays , and RT-PCR analysis of eosinophil mRNA .

<protein>NFAT</protein> expression was determined by using immunoprecipitation and Western blot analysis , DNA-binding assays , and RT-PCR analysis of <rna>eosinophil mRNA</rna> .


RESULTS : Both peripheral blood and bronchoalveolar lavage fluid eosinophils expressed NFATp and NFATc protein .

RESULTS : Both <cell_type>peripheral blood and bronchoalveolar lavage fluid eosinophils</cell_type> expressed <protein>NFATp and NFATc protein</protein> .


Unlike activated T cells , which express multiple NFATc isoforms , eosinophils preferentially express the approximately 85-kd isoform .

Unlike <cell_type>activated T cells</cell_type> , which express multiple NFATc isoforms , <cell_type>eosinophils</cell_type> preferentially express the approximately 85-kd isoform .


In addition , eosinophils were found to constitutively express NFATc mRNA .

In addition , <cell_type>eosinophils</cell_type> were found to constitutively express <rna>NFATc mRNA</rna> .


A brief incubation with the T ( H ) 2 cytokines IL-4 and IL-5 was sufficient to induce the nuclear translocation of NFATc .

A brief incubation with the <protein>T ( H ) 2 cytokines</protein> <protein>IL-4</protein> and <protein>IL-5</protein> was sufficient to induce the nuclear translocation of <protein>NFATc</protein> .


Eosinophil nuclear extracts contain multiple factors that can specifically recognize the IL-4 promoter P1 NFAT site in DNA-binding assays , including NFATp .

Eosinophil nuclear extracts contain multiple factors that can specifically recognize the <dna>IL-4 promoter P1 NFAT site</dna> in DNA-binding assays , including <protein>NFATp</protein> .


CONCLUSION : NFATp and NFATc can regulate the expression of cytokines and other genes in eosinophils but appear to be regulated by a novel signal transduction mechanism in these cells .

CONCLUSION : <protein>NFATp</protein> and <protein>NFATc</protein> can regulate the expression of <protein>cytokines</protein> and other genes in <cell_type>eosinophils</cell_type> but appear to be regulated by a novel signal transduction mechanism in these cells .


BLyS BINDS TO B CELLS WITH HIGH AFFINITY AND INDUCES ACTIVATION OF THE TRANSCRIPTION FACTORS NF-kappaB AND ELF-1 .

<protein>BLyS</protein> BINDS TO <cell_type>B CELLS</cell_type> WITH HIGH AFFINITY AND INDUCES ACTIVATION OF THE <protein>TRANSCRIPTION FACTORS</protein> <protein>NF-kappaB</protein> AND <protein>ELF-1</protein> .


B lymphocyte stimulator ( BLyS ) is a novel member of the TNF family of proteins expressed by myeloid cells as membrane-bound and soluble forms .

<protein>B lymphocyte stimulator</protein> ( <protein>BLyS</protein> ) is a novel member of the <protein>TNF family of proteins</protein> expressed by <cell_type>myeloid cells</cell_type> as membrane-bound and soluble forms .


BLyS was shown to act specifically on B cells , inducing proliferation and immunoglobulin production both in vitro and in vivo .

<protein>BLyS</protein> was shown to act specifically on <cell_type>B cells</cell_type> , inducing proliferation and immunoglobulin production both in vitro and in vivo .


The present study was undertaken to characterize binding of radiolabeled BLyS to its cognate receptor on human B lymphocytes and examine intracellular events initiated by BLyS binding .

The present study was undertaken to characterize binding of <protein>radiolabeled BLyS</protein> to its <protein>cognate receptor</protein> on <cell_type>human B lymphocytes</cell_type> and examine intracellular events initiated by <protein>BLyS</protein> binding .


Similar to other TNF family members , BLyS is present in solution as a homotrimer as determined by gel filtration chromatography and light scattering analysis .

Similar to other <protein>TNF family members</protein> , <protein>BLyS</protein> is present in solution as a <protein>homotrimer</protein> as determined by gel filtration chromatography and light scattering analysis .


BLyS binding to B cells is specific as other TNF family members tested did not compete for ( 125 ) I- BLyS binding .

<protein>BLyS</protein> binding to <cell_type>B cells</cell_type> is specific as other <protein>TNF family members</protein> tested did not compete for ( 125 ) I- <protein>BLyS</protein> binding .


Analysis of equilibrium binding of ( 125 ) I-labeled BLyS to purified human tonsillar B cells demonstrated saturable binding .

Analysis of equilibrium binding of <protein>( 125 ) I-labeled BLyS</protein> to purified <cell_type>human tonsillar B cells</cell_type> demonstrated saturable binding .


Scatchard analysis of the binding data revealed a single class of high-affinity binding on human B cells with approximately 2600 binding sites per cell and an apparent dissociation constant ( K ( D ) ) of about 0.1 nM .

Scatchard analysis of the binding data revealed a single class of high-affinity binding on <cell_type>human B cells</cell_type> with approximately 2600 binding sites per cell and an apparent dissociation constant ( K ( D ) ) of about 0.1 nM .


In addition we report that BLyS binding to B cells results in the activation of NF-kappaB and the Ets family transcription factor , ELF-1 , and in the induction of mRNA for Polo-like kinase ( PLK ) .

In addition we report that <protein>BLyS</protein> binding to <cell_type>B cells</cell_type> results in the activation of <protein>NF-kappaB</protein> and the <protein>Ets family transcription factor</protein> , <protein>ELF-1</protein> , and in the induction of mRNA for <protein>Polo-like kinase</protein> ( <protein>PLK</protein> ) .


Copyright 2001 Academic Press .


Design and use of an inducibly activated human immunodeficiency virus type 1 Nef to study immune modulation .

Design and use of an inducibly activated human immunodeficiency virus type 1 <protein>Nef</protein> to study immune modulation .


The Nef protein of the human immunodeficiency virus type 1 ( HIV-1 ) has been shown to enhance the infectivity of virus particles , downmodulate cell surface proteins , and associate with many intracellular proteins that are thought to facilitate HIV infection .

The <protein>Nef protein</protein> of the human immunodeficiency virus type 1 ( HIV-1 ) has been shown to enhance the infectivity of virus particles , downmodulate <protein>cell surface proteins</protein> , and associate with many <protein>intracellular proteins</protein> that are thought to facilitate HIV infection .


One of the challenges in defining the molecular events regulated by Nef has been obtaining good expression of Nef protein in T cells .

One of the challenges in defining the molecular events regulated by <protein>Nef</protein> has been obtaining good expression of <protein>Nef protein</protein> in <cell_type>T cells</cell_type> .


This has been attributed to effects of Nef on cell proliferation and apoptosis .

This has been attributed to effects of <protein>Nef</protein> on cell proliferation and apoptosis .


We have designed a Nef protein that is readily expressed in T-cell lines and whose function is inducibly activated .

We have designed a <protein>Nef protein</protein> that is readily expressed in <cell_line>T-cell lines</cell_line> and whose function is inducibly activated .


It is composed of a fusion between full-length Nef and the estrogen receptor hormone-binding domain ( Nef-ER ) .

It is composed of a fusion between <protein>full-length Nef</protein> and the <protein>estrogen receptor hormone-binding domain</protein> ( <protein>Nef-ER</protein> ) .


The Nef-ER is kept in an inactive state due to steric hindrance , and addition of the membrane-permeable drug 4-hydroxytamoxifen ( 4-HT ) , which binds to the ER domain , leads to inducible activation of Nef-ER within cells .

The <protein>Nef-ER</protein> is kept in an inactive state due to steric hindrance , and addition of the membrane-permeable drug 4-hydroxytamoxifen ( 4-HT ) , which binds to the <protein>ER domain</protein> , leads to inducible activation of <protein>Nef-ER</protein> within cells .


We demonstrate that Nef-ER inducibly associates with the 62-kDa Ser/Thr kinase and is localized to specific membrane microdomains ( lipid rafts ) only after activation .

We demonstrate that <protein>Nef-ER</protein> inducibly associates with the <protein>62-kDa Ser/Thr kinase</protein> and is localized to specific membrane microdomains ( lipid rafts ) only after activation .


Using this inducible Nef , we also compared the specific requirements for CD4 and HLA-A2 downmodulation in a SupT1 T-cell line .

Using this inducible <protein>Nef</protein> , we also compared the specific requirements for <protein>CD4</protein> and <protein>HLA-A2</protein> downmodulation in a <cell_line>SupT1 T-cell line</cell_line> .