The ML-1 human myeloblastic leukemia cells used in this study proliferate when treated with insulin-like growth factor I and transferrin but differentiate to monocytes when exposed to tumor necrosis factor alpha or transforming growth factor beta1 , or to macrophage-like cells when treated with both these cytokines .

The <cell_line>ML-1 human myeloblastic leukemia cells</cell_line> used in this study proliferate when treated with <protein>insulin-like growth factor I</protein> and <protein>transferrin</protein> but differentiate to <cell_type>monocytes</cell_type> when exposed to <protein>tumor necrosis factor alpha</protein> or <protein>transforming growth factor beta1</protein> , or to <cell_type>macrophage-like cells</cell_type> when treated with both these <protein>cytokines</protein> .


Initiation of proliferation but not of differentiation was followed by a 20- to 25-fold increase in the nuclear level of the DNA polymerase-associated processivity factor PCNA and of the proliferation-specific transcription factor E2F1 .

Initiation of proliferation but not of differentiation was followed by a 20- to 25-fold increase in the nuclear level of the <protein>DNA polymerase-associated processivity factor PCNA</protein> and of the <protein>proliferation-specific transcription factor E2F1</protein> .


In contrast , induction of differentiation but not of proliferation was followed by a 25- to 30-fold increase in the nuclear level of the tumor suppressor proteins p53 ( wild type ) , pRb , and p130/Rb2 and of the p53-dependent cyclin kinase inhibitor p21/Cip1 .

In contrast , induction of differentiation but not of proliferation was followed by a 25- to 30-fold increase in the nuclear level of the <protein>tumor suppressor proteins p53</protein> ( wild type ) , <protein>pRb</protein> , and <protein>p130/Rb2</protein> and of the <protein>p53-dependent cyclin kinase inhibitor</protein> <protein>p21/Cip1</protein> .


p53 and p21/Cip1 , respectively , inhibit the expression and activation of PCNA , whereas p130 and pRb , respectively , inhibit the expression and activation of E2F1 .

<protein>p53</protein> and <protein>p21/Cip1</protein> , respectively , inhibit the expression and activation of <protein>PCNA</protein> , whereas <protein>p130</protein> and <protein>pRb</protein> , respectively , inhibit the expression and activation of <protein>E2F1</protein> .


As a result , G1-S-associated DNA and mRNA synthesis is inhibited , growth uncoupled from differentiation , and maturation enabled to proceed .

As a result , <dna>G1-S-associated DNA</dna> and mRNA synthesis is inhibited , growth uncoupled from differentiation , and maturation enabled to proceed .


Where this function of the tumor suppressor proteins is impaired , the capacity for differentiation is lost , which leads to the sustained proliferation that is characteristic of the cancer cell .

Where this function of the <protein>tumor suppressor proteins</protein> is impaired , the capacity for differentiation is lost , which leads to the sustained proliferation that is characteristic of the <cell_type>cancer cell</cell_type> .


Transcription factor activation in lymphokine activated killer cells and lymphocytes from patients receiving IL-2 immunotherapy .

<protein>Transcription factor</protein> activation in lymphokine activated <cell_type>killer cells</cell_type> and <cell_type>lymphocytes</cell_type> from patients receiving <protein>IL-2</protein> immunotherapy .


Administration of the cytokine interleukin-2 ( IL-2 ) can result in therapeutic benefits for individuals with renal cell carcinoma and melanoma .

Administration of the <protein>cytokine interleukin-2</protein> ( <protein>IL-2</protein> ) can result in therapeutic benefits for individuals with renal cell carcinoma and melanoma .


Here we report an analysis of the transcription factor families AP-1 , Sp1 , NF-kappaB , and signal transducers and activators of transcription ( STAT ) in cancer patients ' lymphocytes before and after IL-2 immunotherapy , as assessed by a gel-shift assay .

Here we report an analysis of the transcription factor families AP-1 , Sp1 , <protein>NF-kappaB</protein> , and <protein>signal transducers and activators of transcription</protein> ( <protein>STAT</protein> ) in <cell_type>cancer patients ' lymphocytes</cell_type> before and after <protein>IL-2</protein> immunotherapy , as assessed by a gel-shift assay .


An in vitro surrogate of IL-2 immunotherapy is the incubation of fresh peripheral blood mononuclear cells ( PBMC ) from healthy individuals in IL-2 for several days , resulting in the production of lymphokine -activated killer ( LAK ) activity in these cultures .

An in vitro surrogate of <protein>IL-2</protein> immunotherapy is the incubation of <cell_type>fresh peripheral blood mononuclear cells</cell_type> ( <cell_type>PBMC</cell_type> ) from healthy individuals in <protein>IL-2</protein> for several days , resulting in the production of <protein>lymphokine</protein> -activated killer ( LAK ) activity in these cultures .


One purpose of this study was to describe the profile of transcription factor activation in these different populations , and assess whether the patterns observed correlated with functional differences in these cells .


Prior to in vivo IL-2 administration , the typical binding pattern of transcription factors in PBMC from patients resembled that seen in fresh PBMC from healthy individuals .

Prior to in vivo <protein>IL-2</protein> administration , the typical binding pattern of <protein>transcription factors</protein> in <cell_type>PBMC</cell_type> from patients resembled that seen in fresh <cell_type>PBMC</cell_type> from healthy individuals .


Over a 3-week course of IL-2 therapy , in most patients the binding patterns of AP-1 , Sp1 , and NF-kappaB proteins changed to resemble those seen in PBMC activated by IL-2 in vitro .

Over a 3-week course of <protein>IL-2</protein> therapy , in most patients the binding patterns of <protein>AP-1</protein> , <protein>Sp1</protein> , and <protein>NF-kappaB</protein> proteins changed to resemble those seen in <cell_type>PBMC</cell_type> activated by <protein>IL-2</protein> in vitro .


However , the cells obtained from IL-2 -treated patients did not have low-level constitutive expression of STAT binding factors as did LAK cells .

However , the cells obtained from <protein>IL-2</protein> -treated patients did not have low-level constitutive expression of <protein>STAT binding factors</protein> as did <protein>LAK cells</protein> .


When these patient cells were further stimulated by IL-2 in vitro , additional differences in STAT induction patterns were noted .

When these patient cells were further stimulated by <protein>IL-2</protein> in vitro , additional differences in <protein>STAT</protein> induction patterns were noted .


These data provide further information on the molecular events occurring in immune cells generated through in vivo and in vitro administration of IL-2 , and further document that there is not a precise congruence between PBMC activated in vivo and in vitro by IL-2 .

These data provide further information on the molecular events occurring in <cell_type>immune cells</cell_type> generated through in vivo and in vitro administration of <protein>IL-2</protein> , and further document that there is not a precise congruence between <cell_type>PBMC</cell_type> activated in vivo and in vitro by <protein>IL-2</protein> .


Activated platelets induce monocyte chemotactic protein-1 secretion and surface expression of intercellular adhesion molecule-1 on endothelial cells [ see comments ]

<cell_type>Activated platelets</cell_type> induce <protein>monocyte chemotactic protein-1</protein> secretion and surface expression of <cell_type>intercellular adhesion molecule-1</cell_type> on <cell_type>endothelial cells</cell_type> [ see comments ]


BACKGROUND : Platelet/endothelium interaction plays an important role in the pathophysiology of inflammation and atherosclerosis .


The role of platelets for monocyte chemotactic protein-1 ( MCP-1 ) secretion and surface expression of intercellular adhesion molecule-1 ( ICAM-1 ) on endothelial cells has been assessed .

The role of platelets for <protein>monocyte chemotactic protein-1</protein> ( <protein>MCP-1</protein> ) secretion and surface expression of <cell_type>intercellular adhesion molecule-1</cell_type> ( <protein>ICAM-1</protein> ) on <cell_type>endothelial cells</cell_type> has been assessed .


METHODS AND RESULTS : Monolayers of human umbilical vein endothelial cells were incubated with nonstimulated or ADP-activated platelets for 6 hours , and secretion of MCP-1 and surface expression of ICAM-1 were determined by ELISA and flow cytometry , respectively .

METHODS AND RESULTS : Monolayers of <cell_type>human umbilical vein endothelial cells</cell_type> were incubated with nonstimulated or <cell_type>ADP-activated platelets</cell_type> for 6 hours , and secretion of <protein>MCP-1</protein> and surface expression of <protein>ICAM-1</protein> were determined by ELISA and flow cytometry , respectively .


In the presence of ADP-activated platelets , both MCP-1 secretion and ICAM-1 surface expression were significantly increased compared with nonstimulated platelets ( P < 0.02 ) .

In the presence of <cell_type>ADP-activated platelets</cell_type> , both <protein>MCP-1</protein> secretion and <protein>ICAM-1</protein> surface expression were significantly increased compared with <cell_type>nonstimulated platelets</cell_type> ( P < 0.02 ) .


Activation of the transcription factor nuclear factor-kappaB ( NF-kappaB ) determined by electrophoretic mobility shift assay and kappaB -dependent transcriptional activity was enhanced in the presence of activated platelets .

Activation of the <protein>transcription factor nuclear factor-kappaB</protein> ( <protein>NF-kappaB</protein> ) determined by electrophoretic mobility shift assay and <protein>kappaB</protein> -dependent transcriptional activity was enhanced in the presence of <cell_type>activated platelets</cell_type> .


In addition , ADP-activated platelets induced MCP-1 and ICAM-1 promoter-dependent transcription .

In addition , <cell_type>ADP-activated platelets</cell_type> induced MCP-1 and ICAM-1 promoter-dependent transcription .


Liposomal transfection of a double-stranded kappaB phosphorothioate oligonucleotide , but not of the mutated form , inhibited MCP-1 secretion and surface expression of ICAM-1 on activated endothelium ( P < 0.05 ) .

Liposomal transfection of a double-stranded kappaB phosphorothioate oligonucleotide , but not of the mutated form , inhibited <protein>MCP-1</protein> secretion and surface expression of <protein>ICAM-1</protein> on activated endothelium ( P < 0.05 ) .


CONCLUSIONS : The present study indicates that activated platelets modulate chemotactic ( MCP-1 ) and adhesive ( ICAM-1 ) properties of endothelial cells via an NF-kappaB -dependent mechanism .

CONCLUSIONS : The present study indicates that <cell_type>activated platelets</cell_type> modulate chemotactic ( <protein>MCP-1</protein> ) and adhesive ( <protein>ICAM-1</protein> ) properties of <cell_type>endothelial cells</cell_type> via an <protein>NF-kappaB</protein> -dependent mechanism .


Platelet-induced activation of the NF-kappaB system might contribute to early inflammatory events in atherogenesis .

Platelet-induced activation of the <protein>NF-kappaB</protein> system might contribute to early inflammatory events in atherogenesis .


Retinoid X receptor and c-cerbA/thyroid hormone receptor regulate erythroid cell growth and differentiation .

<protein>Retinoid X receptor</protein> and <protein>c-cerbA/thyroid hormone receptor</protein> regulate erythroid cell growth and differentiation .


Nuclear receptors are important regulators of erythroid cell development .

<protein>Nuclear receptors</protein> are important regulators of erythroid cell development .


Here we investigated the impact of retinoid X receptor ( RXR ) , retinoic acid receptor ( RAR ) , and of the c-erbA/thyroid hormone ( T3 ) receptor ( c-erbA/TR ) on growth and differentiation of erythroid cells using an in vitro culture system of stem cell factor-dependent erythroid progenitors .

Here we investigated the impact of <protein>retinoid X receptor</protein> ( <protein>RXR</protein> ) , <protein>retinoic acid receptor</protein> ( <protein>RAR</protein> ) , and of the <protein>c-erbA/thyroid hormone ( T3 ) receptor</protein> ( <protein>c-erbA/TR</protein> ) on growth and differentiation of <cell_type>erythroid cells</cell_type> using an in vitro culture system of <cell_type>stem cell factor-dependent erythroid progenitors</cell_type> .


RXR , RAR , and c-erbA/TR -specific ligands were found to induce erythroid-specific gene expression and to accelerate erythroid differentiation in culture , with T3 being most effective .

<protein>RXR</protein> , <protein>RAR</protein> , and <protein>c-erbA/TR</protein> -specific ligands were found to induce <dna>erythroid-specific gene</dna> expression and to accelerate erythroid differentiation in culture , with T3 being most effective .


Furthermore , while ligand-activated c-erbA/TR accelerated differentiation , unliganded c-erbA/TR effectively blocked differentiation and supported sustained progenitor growth in culture .

Furthermore , while ligand-activated <protein>c-erbA/TR</protein> accelerated differentiation , unliganded <protein>c-erbA/TR</protein> effectively blocked differentiation and supported sustained progenitor growth in culture .


Thus , c-erbA/TR appears to act as a binary switch affecting erythroid cell fate : unliganded c-erbA/TR supports growth while ligand-activated c-erbA/TR induces differentiation .

Thus , <protein>c-erbA/TR</protein> appears to act as a binary switch affecting <cell_type>erythroid cell fate</cell_type> : unliganded <protein>c-erbA/TR</protein> supports growth while ligand-activated <protein>c-erbA/TR</protein> induces differentiation .


Additionally , to determine the impact of RXR for erythroid cell development , dominant interfering mutant RXRs , lacking the transcriptional activator functions AF-1 and AF-2 , or AF-2 only , or the entire DNA-binding domain , were introduced into erythroid progenitor cells via recombinant retrovirus vectors and analyzed for RXR -specific effects .

Additionally , to determine the impact of <protein>RXR</protein> for erythroid cell development , dominant interfering <protein>mutant RXRs</protein> , lacking the transcriptional activator functions <protein>AF-1</protein> and <protein>AF-2</protein> , or <protein>AF-2</protein> only , or the <dna>entire DNA-binding domain</dna> , were introduced into <cell_type>erythroid progenitor cells</cell_type> via recombinant retrovirus vectors and analyzed for <protein>RXR</protein> -specific effects .


It was found that expression of wild-type RXR and of the RXR mutants devoid of AF-1 and/or AF-2 supported a transient outgrowth of erythroid cells .

It was found that expression of <protein>wild-type RXR</protein> and of the <protein>RXR mutants</protein> devoid of <protein>AF-1</protein> and/or <protein>AF-2</protein> supported a transient outgrowth of <cell_type>erythroid cells</cell_type> .


In marked contrast , expression of the dominant interfering deltaDNA-binding domain RXR , containing a deletion of the entire DNA-binding domain , was incompatible with erythroid cell growth in vitro , suggesting a pivotal role of RXR for erythroid cell development .

In marked contrast , expression of the <protein>dominant interfering deltaDNA-binding domain</protein> <protein>RXR</protein> , containing a deletion of the <dna>entire DNA-binding domain</dna> , was incompatible with erythroid cell growth in vitro , suggesting a pivotal role of <protein>RXR</protein> for erythroid cell development .


Peripheral T lymphocytes from women with breast cancer exhibit abnormal protein expression of several signaling molecules .

<cell_type>Peripheral T lymphocytes</cell_type> from women with breast cancer exhibit abnormal protein expression of <protein>several signaling molecules</protein> .


We examined signaling molecules of peripheral blood T lymphocytes obtained from women with breast cancer .

We examined signaling molecules of <cell_type>peripheral blood T lymphocytes</cell_type> obtained from women with breast cancer .


In 6 of 14 patients , T lymphocytes displayed an impaired ability to translocate NFeB p65 ( Rel-A ) following activation by anti-CD3 and IL-2 .

In 6 of 14 patients , <cell_type>T lymphocytes</cell_type> displayed an impaired ability to translocate <protein>NFeB p65</protein> ( <protein>Rel-A</protein> ) following activation by anti-CD3 and <protein>IL-2</protein> .


This observation was made despite normal cytoplasmic levels of the Rel-A protein .

This observation was made despite normal cytoplasmic levels of the <protein>Rel-A protein</protein> .


We also detected abnormally low levels of the signaling molecules T-cell receptor ( TCR ) -zeta , ZAP-70 and p56lck in 4 of 14 breast cancer patients , i.e. , defects in T-cell signaling molecules .

We also detected abnormally low levels of the <protein>signaling molecules</protein> <protein>T-cell receptor ( TCR ) -zeta</protein> , <protein>ZAP-70</protein> and <protein>p56lck</protein> in 4 of 14 breast cancer patients , i.e. , defects in <protein>T-cell signaling molecules</protein> .


T lymphocytes from 6 of the 14 patients also exhibited an increased expression of the dual specificity phosphatase , map kinase phosphatase-1 ( MKP-1 ) .

<cell_type>T lymphocytes</cell_type> from 6 of the 14 patients also exhibited an increased expression of the dual <protein>specificity phosphatase</protein> , <protein>map kinase phosphatase-1</protein> ( <protein>MKP-1</protein> ) .


MKP-1 inactivates MAP kinase and therefore may interfere with the activation of c-jun and c-fos .

<protein>MKP-1</protein> inactivates <protein>MAP kinase</protein> and therefore may interfere with the activation of <dna>c-jun</dna> and <dna>c-fos</dna> .


Abnormalities of I or more signaling molecules were found in 9 of 14 patients ; however , only 3 patients had T cells that exhibited all 5 defects .

Abnormalities of I or more <protein>signaling molecules</protein> were found in 9 of 14 patients ; however , only 3 patients had <cell_type>T cells</cell_type> that exhibited all 5 defects .


Our data have implications for the detection of potentially dysfunctional T cells in patients with cancer .

Our data have implications for the detection of potentially dysfunctional <cell_type>T cells</cell_type> in patients with cancer .


For example , the analysis of only 1 signaling molecule may allow patients with significant defects in T-cell signaling to go unnoticed .

For example , the analysis of only 1 <protein>signaling molecule</protein> may allow patients with significant defects in T-cell signaling to go unnoticed .


Finally , despite impaired Rel-A translocation , T cells were capable of transcribing IL-2 .

Finally , despite impaired <protein>Rel-A</protein> translocation , <cell_type>T cells</cell_type> were capable of transcribing <protein>IL-2</protein> .


Impairments in the translocation of Rel-B and c-Rel further suggest that the NFKB family members Rel-A , Rel-B and c-Rel are not required for the transcription of IL-2 in the peripheral T lymphocytes of patients with breast cancer .

Impairments in the translocation of <protein>Rel-B</protein> and <protein>c-Rel</protein> further suggest that the <protein>NFKB family members</protein> <protein>Rel-A</protein> , <protein>Rel-B</protein> and <protein>c-Rel</protein> are not required for the transcription of <protein>IL-2</protein> in the <cell_type>peripheral T lymphocytes</cell_type> of patients with breast cancer .


Specific glucocorticoid binding at different levels of human motor activity .


We studied the number of glucocorticoid receptors and dissociation constant in isolated human lymphocytes as well as blood concentrations of hormones produced by the hypothalamic-hypophyseal-adrenocortical system in three experimental series : at normal ( 17 subjects ) , decreased ( 10 subjects , a 360-d head-down bed rest ) and increased ( 8 subjects , physical exercise on bicycle ergometer ) levels of motor activity .

We studied the number of <protein>glucocorticoid receptors</protein> and dissociation constant in <cell_type>isolated human lymphocytes</cell_type> as well as blood concentrations of hormones produced by the hypothalamic-hypophyseal-adrenocortical system in three experimental series : at normal ( 17 subjects ) , decreased ( 10 subjects , a 360-d head-down bed rest ) and increased ( 8 subjects , physical exercise on bicycle ergometer ) levels of motor activity .


In the first series we found that the number of glucocorticoid receptors and dissociation constant did not depend on the season , on the age of subjects nor on cortisol concentrations in blood .

In the first series we found that the number of <protein>glucocorticoid receptors</protein> and dissociation constant did not depend on the season , on the age of subjects nor on cortisol concentrations in blood .


In the second series we observed the following : at the end of the first month of bed rest the number of glucocorticoid receptors and receptor affinity significantly increased ; at the beginning of the third month of bed rest specific glucocorticoid binding significantly decreased and circadian rhythms of adrenocorticotropin and cortisol in blood varied markedly ; at the end of the sixth month of bed rest the number of glucocorticoid receptors returned to prebed rest levels and dissociation constant decreased .

In the second series we observed the following : at the end of the first month of bed rest the number of <protein>glucocorticoid receptors</protein> and receptor affinity significantly increased ; at the beginning of the third month of bed rest specific glucocorticoid binding significantly decreased and circadian rhythms of adrenocorticotropin and cortisol in blood varied markedly ; at the end of the sixth month of bed rest the number of <protein>glucocorticoid receptors</protein> returned to prebed rest levels and dissociation constant decreased .


In the third series physical exercises that induced an activation of the hypothalamic-hypophyseal-adrenocortical system ( maximal physical exercises and prolonged submaximal exercises at 70 % of maximal oxygen uptake ) led to a significant increase in the number of glucocorticoid receptors without changes of dissociation constant .

In the third series physical exercises that induced an activation of the hypothalamic-hypophyseal-adrenocortical system ( maximal physical exercises and prolonged submaximal exercises at 70 % of maximal oxygen uptake ) led to a significant increase in the number of <protein>glucocorticoid receptors</protein> without changes of dissociation constant .


These results indicate that both a decrease and an increase of human motor activity resulted in significant changes of specific glucocorticoid binding which were not influenced by changes of circulating hormone concentrations in blood but by some other factors affected by physical activity .


The small GTP-binding protein Rho potentiates AP-1 transcription in T cells .

The <protein>small GTP-binding protein Rho</protein> potentiates <protein>AP-1</protein> transcription in <cell_type>T cells</cell_type> .


The Rho family of small GTP-binding proteins is involved in the regulation of cytoskeletal structure , gene transcription , specific cell fate development , and transformation .

The <protein>Rho</protein> family of <protein>small GTP-binding proteins</protein> is involved in the regulation of cytoskeletal structure , gene transcription , specific cell fate development , and transformation .


We demonstrate in this report that overexpression of an activated form of Rho enhances AP-1 activity in Jurkat T cells in the presence of phorbol myristate acetate ( PMA ) , but activated Rho ( V14Rho ) has little or no effect on NFAT , Oct-1 , and NF-kappaB enhancer element activities under similar conditions .

We demonstrate in this report that overexpression of an activated form of <protein>Rho</protein> enhances <protein>AP-1</protein> activity in <cell_line>Jurkat T cells</cell_line> in the presence of phorbol myristate acetate ( PMA ) , but activated <protein>Rho</protein> ( <protein>V14Rho</protein> ) has little or no effect on <protein>NFAT</protein> , <protein>Oct-1</protein> , and <dna>NF-kappaB enhancer element</dna> activities under similar conditions .


Overexpression of a V14Rho construct incapable of membrane localization ( CAAX deleted ) abolishes PMA-induced AP-1 transcriptional activation .

Overexpression of a <dna>V14Rho construct</dna> incapable of membrane localization ( <dna>CAAX deleted</dna> ) abolishes PMA-induced <protein>AP-1</protein> transcriptional activation .


The effect of Rho on AP-1 is independent of the mitogen-activated protein kinase pathway , as a dominant-negative MEK and a MEK inhibitor ( PD98059 ) did not affect Rho -induced AP-1 activity .

The effect of <protein>Rho</protein> on <protein>AP-1</protein> is independent of the <protein>mitogen-activated protein kinase</protein> pathway , as a <protein>dominant-negative MEK</protein> and a <protein>MEK</protein> inhibitor ( PD98059 ) did not affect <protein>Rho</protein> -induced <protein>AP-1</protein> activity .


V14Rho binds strongly to protein kinase Calpha ( PKCalpha ) in vivo ; however , deletion of the CAAX site on V14Rho severely diminished this association .

<protein>V14Rho</protein> binds strongly to <protein>protein kinase Calpha</protein> ( <protein>PKCalpha</protein> ) in vivo ; however , deletion of the <dna>CAAX site</dna> on <protein>V14Rho</protein> severely diminished this association .


Evidence for a role for PKCalpha as an effector of Rho was obtained by the observation that coexpression of the N-terminal domain of PKCalpha blocked the effects of activated Rho plus PMA on AP-1 transcriptional activity .

Evidence for a role for <protein>PKCalpha</protein> as an effector of <protein>Rho</protein> was obtained by the observation that coexpression of the <protein>N-terminal domain</protein> of <protein>PKCalpha</protein> blocked the effects of activated <protein>Rho</protein> plus PMA on <protein>AP-1</protein> transcriptional activity .


These data suggest that Rho potentiates AP-1 transcription during T-cell activation

These data suggest that <protein>Rho</protein> potentiates <protein>AP-1</protein> transcription during T-cell activation


Changes in PKC isoforms in human alveolar macrophages compared with blood monocytes .

Changes in <protein>PKC isoforms</protein> in <cell_type>human alveolar macrophages</cell_type> compared with <cell_type>blood monocytes</cell_type> .


Alveolar macrophages play an important role in host defense and in other types of inflammatory processes in the lung .


These cells exhibit many alterations in function compared with their precursor cells , blood monocytes .

These cells exhibit many alterations in function compared with their <cell_type>precursor cells</cell_type> , <cell_type>blood monocytes</cell_type> .


To evaluate a potential mechanism for these differences in function , we evaluated expression of protein kinase C ( PKC ) isoforms .

To evaluate a potential mechanism for these differences in function , we evaluated expression of <protein>protein kinase C ( PKC ) isoforms</protein> .


We found an increase in Ca2+-dependent PKC isoforms in monocytes compared with alveolar macrophages .

We found an increase in <protein>Ca2+-dependent PKC isoforms</protein> in monocytes compared with alveolar macrophages .


We also found differential expression of the Ca2+-independent isoforms in alveolar macrophages compared with monocytes .

We also found differential expression of the <protein>Ca2+-independent isoforms</protein> in <cell_type>alveolar macrophages</cell_type> compared with <cell_type>monocytes</cell_type> .


One consequence of the activation of PKC can be increased expression of mitogen-activated protein ( MAP ) kinase pathways .

One consequence of the activation of <protein>PKC</protein> can be increased expression of <protein>mitogen-activated protein ( MAP ) kinase</protein> pathways .


Therefore , we also evaluated activation of the MAP kinase extracellular signal-regulated kinase ( ERK ) 2 by the phorbol ester phorbol 12-myristate 13-acetate ( PMA ) .

Therefore , we also evaluated activation of the <protein>MAP kinase extracellular signal-regulated kinase ( ERK ) 2</protein> by the phorbol ester phorbol 12-myristate 13-acetate ( PMA ) .


PMA activated ERK2 kinase in both alveolar macrophages and monocytes ; however , monocytes consistently showed a significantly greater activation of ERK2 kinase by PMA compared with alveolar macrophages .

PMA activated <protein>ERK2 kinase</protein> in both <cell_type>alveolar macrophages</cell_type> and <cell_type>monocytes</cell_type> ; however , <cell_type>monocytes</cell_type> consistently showed a significantly greater activation of <protein>ERK2 kinase</protein> by PMA compared with <cell_type>alveolar macrophages</cell_type> .


Another known consequence of the activation of PKC and subsequent activation of ERK kinase is activation of the transcription factor activator protein-1 ( AP-1 ) .

Another known consequence of the activation of <protein>PKC</protein> and subsequent activation of <protein>ERK kinase</protein> is activation of the <protein>transcription factor activator protein-1</protein> ( <protein>AP-1</protein> ) .


We evaluated the activation of AP-1 by PMA in both monocytes and macrophages .

We evaluated the activation of <protein>AP-1</protein> by PMA in both <cell_type>monocytes</cell_type> and macrophages .


We found very little detectable activation of AP-1 , as assessed in a gel shift assay , in alveolar macrophages , whereas monocytes showed a substantial activation of AP-1 by PMA .

We found very little detectable activation of <protein>AP-1</protein> , as assessed in a gel shift assay , in <cell_type>alveolar macrophages</cell_type> , whereas <cell_type>monocytes</cell_type> showed a substantial activation of <protein>AP-1</protein> by PMA .


These studies show that the differential expression of PKC isoforms in alveolar macrophages and blood monocytes is associated with important functional alterations in the cells .

These studies show that the differential expression of <protein>PKC isoforms</protein> in <cell_type>alveolar macrophages</cell_type> and <cell_type>blood monocytes</cell_type> is associated with important functional alterations in the cells .


Expression of gamma-IFN responsive genes in scavenger receptor over-expressing monocytes is associated with xanthomatosis .

Expression of <dna>gamma-IFN responsive genes</dna> in scavenger receptor <cell_type>over-expressing monocytes</cell_type> is associated with xanthomatosis .


We have recently described an inherited over-expression of the macrophage scavenger receptor ( SR ) in blood monocytes from members of a kindred , only two of whom displayed extensive xanthomatosis .

We have recently described an inherited over-expression of the <protein>macrophage scavenger receptor</protein> ( <protein>SR</protein> ) in <cell_type>blood monocytes</cell_type> from members of a kindred , only two of whom displayed extensive xanthomatosis .


Using mRNA differential display we demonstrated abnormally high expression of the signal transducer and activator of transcription ( STAT1alpha ) in monocytes from the proband II-2 .

Using mRNA differential display we demonstrated abnormally high expression of the <protein>signal transducer and activator of transcription</protein> ( <protein>STAT1alpha</protein> ) in <cell_type>monocytes</cell_type> from the <cell_line>proband II-2</cell_line> .


Expression of gamma-interferon inducible protein 10 ( IP-10 ) , a STAT1alpha-responsive gene and mediator of inflammatory response , was also abnormally expressed in the monocytes from II-2 .

Expression of <protein>gamma-interferon inducible protein 10</protein> ( <protein>IP-10</protein> ) , a <dna>STAT1alpha-responsive gene</dna> and mediator of inflammatory response , was also abnormally expressed in the <cell_type>monocytes</cell_type> from <cell_line>II-2</cell_line> .


Over-expression of both genes was restricted to monocytes from II-2 and was not observed in monocytes from the clinically unaffected family members , unlike that of SR .

Over-expression of both genes was restricted to <cell_type>monocytes</cell_type> from <cell_line>II-2</cell_line> and was not observed in <cell_type>monocytes</cell_type> from the clinically unaffected family members , unlike that of <protein>SR</protein> .


Gel retardation assays with THP-1 cell extracts identified gamma-IFN inducible DNA binding activity to three potential STATI DNA binding elements in the human IP-10 promoter region from nucleotides - 245 to - 188 .

Gel retardation assays with <cell_line>THP-1 cell</cell_line> extracts identified <protein>gamma-IFN</protein> inducible DNA binding activity to three potential <dna>STATI DNA binding elements</dna> in the <dna>human IP-10 promoter region</dna> from <dna>nucleotides - 245 to - 188</dna> .


Taken together these results suggest that gamma-interferon mediated cell activation is responsible for STAT1alpha -induced transcription of the IP-10 gene in THP-1 macrophages as well as in monocytes from II-2 .

Taken together these results suggest that gamma-interferon mediated cell activation is responsible for <protein>STAT1alpha</protein> -induced transcription of the <protein>IP-10</protein> gene in <cell_line>THP-1 macrophages</cell_line> as well as in <cell_type>monocytes</cell_type> from <cell_line>II-2</cell_line> .


Analysis of monocytes from familial hypercholesterolemic ( FH ) subjects , who frequently develop xanthomatosis , revealed a significant number of subjects with elevated STAT1alpha and IP-10 expression .

Analysis of <cell_type>monocytes</cell_type> from familial hypercholesterolemic ( FH ) subjects , who frequently develop xanthomatosis , revealed a significant number of subjects with elevated STAT1alpha and IP-10 expression .


Our data suggest that the inflammatory effects of gamma-IFN signaling could play a role in foam cell formation and xanthomatosis .

Our data suggest that the inflammatory effects of <protein>gamma-IFN</protein> signaling could play a role in foam cell formation and xanthomatosis .


Redox signals and NF-kappaB activation in T cells .

Redox signals and <protein>NF-kappaB</protein> activation in <cell_type>T cells</cell_type> .


Accumulating data from a number of laboratories have recently indicated that the response of transcription factor NF-kappaB to alterations in the redox homeostasis of cells may play an important role in modulating immune function .

Accumulating data from a number of laboratories have recently indicated that the response of <protein>transcription factor</protein> <protein>NF-kappaB</protein> to alterations in the redox homeostasis of cells may play an important role in modulating immune function .


The activation of NF-kappaB has been recognized to regulate a number of genes necessary for normal T cell responses including IL-2 , IL-6 , IL-8 , and several T cell surface receptors .

The activation of <protein>NF-kappaB</protein> has been recognized to regulate a number of genes necessary for normal T cell responses including <protein>IL-2</protein> , <protein>IL-6</protein> , <protein>IL-8</protein> , and several <protein>T cell surface receptors</protein> .


Diminished NF-kappaB activity has been shown to occur in T cells with aging , suggesting that impaired activation of NF-kappaB might occur during cellular senescence .

Diminished <protein>NF-kappaB</protein> activity has been shown to occur in <cell_type>T cells</cell_type> with aging , suggesting that impaired activation of <protein>NF-kappaB</protein> might occur during cellular senescence .


In addition , aberrancies in NF-kappaB activity have been implicated in the immunopathogenesis of diseases involving immune or inflammatory processes such as atherosclerosis and HIV-1 infection .

In addition , aberrancies in <protein>NF-kappaB</protein> activity have been implicated in the immunopathogenesis of diseases involving immune or inflammatory processes such as atherosclerosis and HIV-1 infection .


The role of H2O2 and other reactive oxygen species ( ROS ) as an integratory secondary messenger for divergent T cell signals has been complicated by the fact that various T cell lines and peripheral blood T cells differ markedly in the levels of NF-kappaB activation induced by oxidant stress .

The role of H2O2 and other reactive oxygen species ( ROS ) as an integratory secondary messenger for divergent T cell signals has been complicated by the fact that various <cell_line>T cell lines</cell_line> and <cell_type>peripheral blood T cells</cell_type> differ markedly in the levels of <protein>NF-kappaB</protein> activation induced by oxidant stress .


Additionally , proposed pathways of NF-kappaB activation have been based on indirect evidence provided by experiments which used antioxidants to inhibit active NF-kappaB formation .

Additionally , proposed pathways of <protein>NF-kappaB</protein> activation have been based on indirect evidence provided by experiments which used antioxidants to inhibit active <protein>NF-kappaB</protein> formation .


Further , complete activation of T cells requires at least two signals , one that stimulates an increase in intracellular calcium and one that stimulates enzymatic processes including kinases .

Further , complete activation of <cell_type>T cells</cell_type> requires at least two signals , one that stimulates an increase in intracellular calcium and one that stimulates enzymatic processes including kinases .


Similarly , substantial evidence indicates that full activation of NF-kappaB requires dual signals .

Similarly , substantial evidence indicates that full activation of <protein>NF-kappaB</protein> requires dual signals .


The ability of H2O2 or other ROS to induce T cell signals and functional responses by these two mechanisms is reviewed and the specific response of NF-kappaB to redox changes in T cells is examined .

The ability of H2O2 or other ROS to induce T cell signals and functional responses by these two mechanisms is reviewed and the specific response of <protein>NF-kappaB</protein> to redox changes in <cell_type>T cells</cell_type> is examined .


Data are also presented to suggest that the redox regulation in NF-kappaB activation may be relevant to immune-related diseases and to aging .

Data are also presented to suggest that the redox regulation in <protein>NF-kappaB</protein> activation may be relevant to immune-related diseases and to aging .


Uncoupling of nonreceptor tyrosine kinases from PLC-gamma1 in an SLP-76-deficient T cell .

Uncoupling of nonreceptor <protein>tyrosine kinases</protein> from <protein>PLC-gamma1</protein> in an SLP-76-deficient T cell .


Activation of nonreceptor protein tyrosine kinases ( PTKs ) is essential for T cell receptor ( TCR ) responsiveness ; however , the function of individual PTK substrates is often uncertain .

Activation of <protein>nonreceptor protein tyrosine kinases</protein> ( <protein>PTKs</protein> ) is essential for <protein>T cell receptor</protein> ( <protein>TCR</protein> ) responsiveness ; however , the function of individual <protein>PTK substrates</protein> is often uncertain .


A mutant T cell line was isolated that lacked expression of SLP-76 ( SH2 domain-containing leukocyte protein of 76 kilodaltons ) , a hematopoietically expressed adaptor protein and PTK substrate .

A <cell_line>mutant T cell line</cell_line> was isolated that lacked expression of <protein>SLP-76</protein> ( <protein>SH2 domain-containing leukocyte protein</protein> of 76 kilodaltons ) , a <protein>hematopoietically expressed adaptor protein</protein> and <protein>PTK substrate</protein> .


SLP-76 was not required for TCR -induced tyrosine phosphorylation of most proteins , but was required for optimal tyrosine phosphorylation and activation of phospholipase C-gamma1 ( PLC-gamma1 ) , as well as Ras pathway activation .

<protein>SLP-76</protein> was not required for <protein>TCR</protein> -induced tyrosine phosphorylation of most proteins , but was required for optimal tyrosine phosphorylation and activation of <protein>phospholipase C-gamma1</protein> ( <protein>PLC-gamma1</protein> ) , as well as <protein>Ras</protein> pathway activation .


TCR -inducible gene expression was dependent on SLP-76 .

<protein>TCR</protein> -inducible gene expression was dependent on <protein>SLP-76</protein> .


Thus , coupling of TCR-regulated PTKs to downstream signaling pathways requires SLP-76 .

Thus , coupling of <protein>TCR-regulated PTKs</protein> to downstream signaling pathways requires <protein>SLP-76</protein> .


A novel function of Stat1 and Stat3 proteins in erythropoietin -induced erythroid differentiation of a human leukemia cell line .

A novel function of <protein>Stat1</protein> and <protein>Stat3</protein> proteins in <protein>erythropoietin</protein> -induced erythroid differentiation of a <cell_line>human leukemia cell line</cell_line> .


We recently determined that erythropoietin ( EPO ) activates 3 members of the signal transducer and activator of transcription ( STAT ) family , Stat1alpha , Stat3 , and Stat5 , in the human EPO-dependent cell lines , UT-7 and UT-7/EPO ( Kirito et al , J Biol Chem 272 : 16507 , 1997 ) .

We recently determined that <protein>erythropoietin</protein> ( <protein>EPO</protein> ) activates 3 members of the signal transducer and activator of transcription ( STAT ) family , <protein>Stat1alpha</protein> , <protein>Stat3</protein> , and <protein>Stat5</protein> , in the <cell_line>human EPO-dependent cell lines</cell_line> , <cell_line>UT-7</cell_line> and <cell_line>UT-7/EPO</cell_line> ( Kirito et al , J Biol Chem 272 : 16507 , 1997 ) .


In addition , we have shown that Stat1alpha , but not Stat3 , is involved in EPO -induced cellular proliferation .

In addition , we have shown that <protein>Stat1alpha</protein> , but not <protein>Stat3</protein> , is involved in <protein>EPO</protein> -induced cellular proliferation .


In this study , we examined the roles of Stat1alpha and Stat3 in EPO -induced erythroid differentiation .

In this study , we examined the roles of <protein>Stat1alpha</protein> and <protein>Stat3</protein> in <protein>EPO</protein> -induced erythroid differentiation .


UT-7/GM was used as a model system , because this cell line can differentiate into erythroid-lineage cells with EPO treatment ( Komatsu et al , Blood 89 : 4021 , 1997 ) .

<cell_line>UT-7/GM</cell_line> was used as a model system , because this cell line can differentiate into <cell_type>erythroid-lineage cells</cell_type> with <protein>EPO</protein> treatment ( Komatsu et al , Blood 89 : 4021 , 1997 ) .


We found that EPO did not activate Stat1alpha or Stat3 in UT-7/GM cells .

We found that <protein>EPO</protein> did not activate <protein>Stat1alpha</protein> or <protein>Stat3</protein> in <cell_line>UT-7/GM cells</cell_line> .


Transfection experiments showed that both Stat1alpha and Stat3 inhibited the induction by EPO of gamma-globin and erythroid-specific 5-aminolevulinate synthetase transcripts , resulting in a reduction of the percentage of hemoglobin-positive cells .

Transfection experiments showed that both <protein>Stat1alpha</protein> and <protein>Stat3</protein> inhibited the induction by <protein>EPO</protein> of <rna>gamma-globin and erythroid-specific 5-aminolevulinate synthetase transcripts</rna> , resulting in a reduction of the percentage of <cell_type>hemoglobin-positive cells</cell_type> .


Dominant negative forms of Stat1alpha or Stat3 promoted the EPO -induced erythroid differentiation of UT-7/GM cells , even in the presence of granulocyte-macrophage colony-stimulating factor , although this cytokine never induced erythroid differentiation of the parent UT-7/GM cells with or without EPO .

Dominant negative forms of <protein>Stat1alpha</protein> or <protein>Stat3</protein> promoted the <protein>EPO</protein> -induced erythroid differentiation of <cell_line>UT-7/GM cells</cell_line> , even in the presence of <protein>granulocyte-macrophage colony-stimulating factor</protein> , although this cytokine never induced erythroid differentiation of the <cell_line>parent UT-7/GM cells</cell_line> with or without <protein>EPO</protein> .


A cell cycle analysis showed that the constitutive activation of Stat1alpha , but not Stat3 , shortened the period of G0/G1 prolongation caused by EPO stimulation .

A cell cycle analysis showed that the constitutive activation of <protein>Stat1alpha</protein> , but not <protein>Stat3</protein> , shortened the period of G0/G1 prolongation caused by <protein>EPO</protein> stimulation .


Taken together , our data suggest that Stat1alpha and Stat3 act as negative regulators in EPO -induced erythroid differentiation .

Taken together , our data suggest that <protein>Stat1alpha</protein> and <protein>Stat3</protein> act as negative regulators in <protein>EPO</protein> -induced erythroid differentiation .


Specifically , Stat1alpha may activate a cell cycle-associated gene ( s ) , leading to the entry of cells into the cell cycle .

Specifically , <protein>Stat1alpha</protein> may activate a <dna>cell cycle-associated gene</dna> ( s ) , leading to the entry of cells into the cell cycle .


Ro 09-2210 exhibits potent anti-proliferative effects on activated T cells by selectively blocking MKK activity .

Ro 09-2210 exhibits potent anti-proliferative effects on activated <cell_type>T cells</cell_type> by selectively blocking <protein>MKK</protein> activity .


By using high throughput screening of microbial broths , we have identified a compound , designated Ro 09-2210 , which is able to block anti-CD3 induced peripheral blood T cell activation with an IC50 = 40 nM .

By using high throughput screening of microbial broths , we have identified a compound , designated Ro 09-2210 , which is able to block <protein>anti-CD3</protein> induced peripheral blood T cell activation with an IC50 = 40 nM .


Ro 09-2210 was also able to block antigen-induced IL-2 secretion with an IC50 = 30 nM , but was considerably less potent at blocking Ca2+ flux stimulated by anti-CD3 treatment .

Ro 09-2210 was also able to block antigen-induced <protein>IL-2</protein> secretion with an IC50 = 30 nM , but was considerably less potent at blocking Ca2+ flux stimulated by <protein>anti-CD3</protein> treatment .


To determine the mechanism of action of Ro 09-2210 , we set up a transient expression system in Jurkat T cells using a variety of reporter gene constructs and showed effective inhibition of phorbol ester/ionomycin-induced NF-AT activation and anti-CD3 induced NF-AT with IC50 = 7.7 and 10 nM , respectively .

To determine the mechanism of action of Ro 09-2210 , we set up a transient expression system in <cell_line>Jurkat T cells</cell_line> using a variety of reporter gene constructs and showed effective inhibition of phorbol ester/ionomycin-induced <protein>NF-AT</protein> activation and anti-CD3 induced <protein>NF-AT</protein> with IC50 = 7.7 and 10 nM , respectively .


Ro 09-2210 was also able to inhibit phorbol ester/ionomycin-induced activation of AP1 with IC50 = < 10 nM .

Ro 09-2210 was also able to inhibit phorbol ester/ionomycin-induced activation of <protein>AP1</protein> with IC50 = < 10 nM .


We further showed that Ro 09-2210 was unable to inhibit c-jun induced expression of AP1-dependent reporter constructs ( IC50 > 500 nM ) , but was able to potently inhibit ras-induced AP1 activation ( IC50 = 20 nM ) .

We further showed that Ro 09-2210 was unable to inhibit <dna>c-jun</dna> induced expression of <dna>AP1-dependent reporter constructs</dna> ( IC50 > 500 nM ) , but was able to potently inhibit ras-induced <protein>AP1</protein> activation ( IC50 = 20 nM ) .


This suggested that Ro 09-2210 was inhibiting an activator of AP-1 which was upstream of c-jun and downstream of ras signaling .

This suggested that Ro 09-2210 was inhibiting an activator of <protein>AP-1</protein> which was upstream of <dna>c-jun</dna> and downstream of <protein>ras</protein> signaling .


To investigate further , we then purified a number of different kinases , including PKC , PhK , ZAP-70 , ERK , and MEK 1 ( a MKK ) , and showed that Ro 09-2210 was a selective inhibitor of MEK1 in vitro ( IC50 = 59 nM ) .

To investigate further , we then purified a number of different kinases , including <protein>PKC</protein> , <protein>PhK</protein> , <protein>ZAP-70</protein> , <protein>ERK</protein> , and <protein>MEK 1</protein> ( a <protein>MKK</protein> ) , and showed that Ro 09-2210 was a selective inhibitor of <protein>MEK1</protein> in vitro ( IC50 = 59 nM ) .


Coactivation by OCA-B : definition of critical regions and synergism with general cofactors .

Coactivation by <protein>OCA-B</protein> : definition of critical regions and synergism with general cofactors .


Molecular dissection of the B-cell-specific transcription coactivator OCA-B has revealed distinct regions important , respectively , for recruitment to immunoglobulin promoters through interaction with octamer-bound Oct-1 and for subsequent coactivator function .

Molecular dissection of the B-cell-specific transcription coactivator <protein>OCA-B</protein> has revealed distinct regions important , respectively , for recruitment to <dna>immunoglobulin promoters</dna> through interaction with <protein>octamer-bound Oct-1</protein> and for subsequent coactivator function .


Further analysis of general coactivator requirements showed that selective removal of PC4 from the essential USA fraction severely impairs Oct-1 and OCA-B function in a cell-free system reconstituted with partially purified factors .

Further analysis of general coactivator requirements showed that selective removal of <protein>PC4</protein> from the essential USA fraction severely impairs Oct-1 and OCA-B function in a cell-free system reconstituted with partially purified factors .


Full activity can be restored by the combined action of recombinant PC4 and the PC4 -depleted USA fraction , thus suggesting a joint requirement for PC4 and another , USA-derived component ( s ) for optimal function of Oct-1 /OCA-B in the reconstituted system .

Full activity can be restored by the combined action of recombinant <protein>PC4</protein> and the <protein>PC4</protein> -depleted USA fraction , thus suggesting a joint requirement for <protein>PC4</protein> and another , USA-derived component ( s ) for optimal function of <protein>Oct-1</protein> <protein>/OCA-B</protein> in the reconstituted system .


Indeed , USA-derived PC2 was found to act synergistically with PC4 in reproducing the function of intact USA in the assay system .

Indeed , <protein>USA-derived PC2</protein> was found to act synergistically with <protein>PC4</protein> in reproducing the function of intact USA in the assay system .


Consistent with the requirement for PC4 in the reconstituted system , OCA-B was found to interact directly with PC4 .

Consistent with the requirement for <protein>PC4</protein> in the reconstituted system , <protein>OCA-B</protein> was found to interact directly with <protein>PC4</protein> .


Surprisingly , however , removal of PC4 from the unfractionated nuclear extract has no detrimental effect on OCA-B / Oct-1 -dependent transcription .

Surprisingly , however , removal of <protein>PC4</protein> from the unfractionated nuclear extract has no detrimental effect on <protein>OCA-B</protein> / <protein>Oct-1</protein> -dependent transcription .


These results lead to a general model for the synergistic function of activation domains in Oct-1 and OCA-B ( mediated by the combined action of the multiple USA components ) and , further , suggest a functional redundancy in general coactivators .

These results lead to a general model for the synergistic function of activation domains in <protein>Oct-1</protein> and <protein>OCA-B</protein> ( mediated by the combined action of the multiple USA components ) and , further , suggest a functional redundancy in <protein>general coactivators</protein> .


Activation of Stat-3 is involved in the induction of apoptosis after ligation of major histocompatibility complex class I molecules on human Jurkat T cells .

Activation of <protein>Stat-3</protein> is involved in the induction of apoptosis after ligation of <protein>major histocompatibility complex class I molecules</protein> on <cell_line>human Jurkat T cells</cell_line> .


Activation of Janus tyrosine kinases ( Jak ) and Signal transducers and activators of transcription ( Stat ) after ligation of major histocompatibility complex class I ( MHC-I ) was explored in Jurkat T cells .

Activation of <protein>Janus tyrosine kinases</protein> ( <protein>Jak</protein> ) and <protein>Signal transducers and activators of transcription</protein> ( <protein>Stat</protein> ) after ligation of <protein>major histocompatibility complex class I</protein> ( <protein>MHC-I</protein> ) was explored in <cell_line>Jurkat T cells</cell_line> .


Cross-linking of MHC-I mediated tyrosine phosphorylation of Tyk2 , but not Jak1 , Jak2 , and Jak3 .

Cross-linking of <protein>MHC-I</protein> mediated tyrosine phosphorylation of <protein>Tyk2</protein> , but not <protein>Jak1</protein> , <protein>Jak2</protein> , and <protein>Jak3</protein> .


In addition , the transcription factor Stat-3 was tyrosine phosphorylated in the cytoplasm and subsequently translocated to the cell nucleus .

In addition , the <protein>transcription factor</protein> <protein>Stat-3</protein> was tyrosine phosphorylated in the cytoplasm and subsequently translocated to the cell nucleus .


Data obtained by electrophoretic mobility shift assay suggested that the activated Stat-3 protein associates with the human serum-inducible element ( hSIE ) DNA-probe derived from the interferon-gamma activated site ( GAS ) in the c-fos promoter , a common DNA sequence for Stat protein binding .

Data obtained by electrophoretic mobility shift assay suggested that the <protein>activated Stat-3 protein</protein> associates with the <dna>human serum-inducible element ( hSIE ) DNA-probe</dna> derived from the <dna>interferon-gamma activated site</dna> ( <dna>GAS</dna> ) in the <dna>c-fos promoter</dna> , a common DNA sequence for <protein>Stat protein</protein> binding .


An association between hSIE and Stat-3 after MHC-I ligation was directly demonstrated by precipitating Stat-3 from nuclear extracts with biotinylated hSIE probe and avidin -coupled agarose .

An association between <dna>hSIE</dna> and <protein>Stat-3</protein> after <protein>MHC-I</protein> ligation was directly demonstrated by precipitating <protein>Stat-3</protein> from nuclear extracts with biotinylated <dna>hSIE probe</dna> and <protein>avidin</protein> -coupled agarose .


To investigate the function of the activated Stat-3 , Jurkat T cells were transiently transfected with a Stat-3 isoform lacking the transactivating domain .

To investigate the function of the activated <protein>Stat-3</protein> , <cell_line>Jurkat T cells</cell_line> were transiently transfected with a <protein>Stat-3 isoform</protein> lacking the <protein>transactivating domain</protein> .


This dominant-negative acting Stat-3 isoform significantly inhibited apoptosis induced by ligation of MHC-I .

This <protein>dominant-negative acting Stat-3 isoform</protein> significantly inhibited apoptosis induced by ligation of <protein>MHC-I</protein> .


In conclusion , our data suggest the involvement of the Jak/Stat signal pathway in MHC-I -induced signal transduction in T cells .

In conclusion , our data suggest the involvement of the <protein>Jak/Stat</protein> signal pathway in <protein>MHC-I</protein> -induced signal transduction in <cell_type>T cells</cell_type> .


IL-2 -induced growth of CD8+ T cell prolymphocytic leukemia cells mediated by NF-kappaB induction and IL-2 receptor alpha expression .

<protein>IL-2</protein> -induced growth of <cell_type>CD8+ T cell prolymphocytic leukemia cells</cell_type> mediated by <protein>NF-kappaB</protein> induction and <protein>IL-2 receptor alpha</protein> expression .


The binding of interleukin-2 ( IL-2 ) to its receptor on normal T cells induces nuclear expression of nuclear factor kappaB ( NF-kappaB ) , activation of the IL-2 receptor ( IL-2R ) alpha chain gene , and cell proliferation .

The binding of <protein>interleukin-2</protein> ( <protein>IL-2</protein> ) to its receptor on normal <cell_type>T cells</cell_type> induces nuclear expression of <protein>nuclear factor kappaB</protein> ( <protein>NF-kappaB</protein> ) , activation of the <dna>IL-2 receptor ( IL-2R ) alpha chain gene</dna> , and cell proliferation .


In the present study , the role of IL-2R signaling in the growth of CD8+ T cell prolymphocytic leukemia ( T-PLL ) cells has been investigated .

In the present study , the role of <protein>IL-2R</protein> signaling in the growth of <cell_line>CD8+ T cell prolymphocytic leukemia ( T-PLL ) cells</cell_line> has been investigated .


Flow cytometry revealed that primary leukemia cells from a patient with CD8+ T-PLL expressed IL-2Ralpha and beta chains , and the cells showed a proliferative response and an increase in IL-2Ralpha expression on culture with exogeneous IL-2 .

Flow cytometry revealed that <cell_type>primary leukemia cells</cell_type> from a patient with CD8+ T-PLL expressed <protein>IL-2Ralpha and beta chains</protein> , and the cells showed a proliferative response and an increase in <protein>IL-2Ralpha</protein> expression on culture with exogeneous <protein>IL-2</protein> .


Northern blot analysis failed to detect IL-2 mRNA , suggesting that IL-2 may act in a paracrine manner in vivo .

Northern blot analysis failed to detect <rna>IL-2 mRNA</rna> , suggesting that <protein>IL-2</protein> may act in a paracrine manner in vivo .


Electrophoretic mobility-shift assays revealed that recombinant IL-2 increased NF-kappaB binding activity in nuclear extracts of the leukemia cells , and Northern blot analysis showed that IL-2 increased the abundance of mRNAs encoding the NF-kappaB components c-Rel and KBF1 in these cells .

Electrophoretic mobility-shift assays revealed that recombinant <protein>IL-2</protein> increased <protein>NF-kappaB</protein> binding activity in nuclear extracts of the <cell_type>leukemia cells</cell_type> , and Northern blot analysis showed that <protein>IL-2</protein> increased the abundance of mRNAs encoding the <protein>NF-kappaB</protein> components <protein>c-Rel</protein> and <protein>KBF1</protein> in these cells .


IL-2 binding analysis demonstrated that IL-2 markedly increased the number of low affinity IL-2Rs on the leukemia cells , without an effect on the number of high-affinity IL-2Rs .

<protein>IL-2</protein> binding analysis demonstrated that <protein>IL-2</protein> markedly increased the number of low affinity <protein>IL-2Rs</protein> on the <cell_type>leukemia cells</cell_type> , without an effect on the number of <protein>high-affinity IL-2Rs</protein> .


These results show that IL-2 is capable of inducing the nuclear expression of NF-kappaB in primary CD8+ T-PLL cells , and that this effect is mediated , at least in part , at a pretranslational level

These results show that <protein>IL-2</protein> is capable of inducing the nuclear expression of <protein>NF-kappaB</protein> in <cell_line>primary CD8+ T-PLL cells</cell_line> , and that this effect is mediated , at least in part , at a pretranslational level


Dimethyldithiocarbamate inhibits in vitro activation of primary human CD4+ T lymphocytes .

Dimethyldithiocarbamate inhibits in vitro activation of <cell_type>primary human CD4+ T lymphocytes</cell_type> .


Dithiocarbamates ( DTC ) , a diverse group of industrial and therapeutic chemicals , have been reported to inhibit , enhance or have no effect on the immune system .


These apparent inconsistencies reflect the complexity of the DTCs biological activities and are probably due in part to differences in dose , route of exposure , animal species used and/or specific compound tested .


The studies described herein were undertaken to investigate the immunotoxicity of one member of this family , dimethyldithiocarbamate ( DMDTC ) .


We demonstrate that 0.1-0.5 microM DMDTC inhibits TNF-alpha -induced activation of NF-kappaB in primary human CD4+ T cells .

We demonstrate that 0.1-0.5 microM DMDTC inhibits <protein>TNF-alpha</protein> -induced activation of <protein>NF-kappaB</protein> in <cell_type>primary human CD4+ T cells</cell_type> .


This inhibition is not accompanied by a loss in viability , and DMDTC-treated T cells retain other active signaling pathways throughout the exposure duration .

This inhibition is not accompanied by a loss in viability , and <cell_type>DMDTC-treated T cells</cell_type> retain other active signaling pathways throughout the exposure duration .


The inhibition of NF-kappaB is apparently permanent as DMDTC-treated T cells did not regain normal TNF-alpha activation , even after 72 h in culture .

The inhibition of <protein>NF-kappaB</protein> is apparently permanent as <cell_type>DMDTC-treated T cells</cell_type> did not regain normal <protein>TNF-alpha</protein> activation , even after 72 h in culture .


DMDTC does not appear to alter NF-kappaB directly as pre-incubation of nuclear extracts with DMDTC does not diminish binding activity of this protein .

DMDTC does not appear to alter <protein>NF-kappaB</protein> directly as pre-incubation of nuclear extracts with DMDTC does not diminish binding activity of this protein .


We further demonstrate that 0.1-0.5 microM DMDTC inhibits intracellular IL-2 production and decreases surface expression of CD25 ( the alpha subunit of the IL-2 receptor ) in T cells stimulated with phorbol ester .

We further demonstrate that 0.1-0.5 microM DMDTC inhibits intracellular <protein>IL-2</protein> production and decreases surface expression of <protein>CD25</protein> ( the <protein>alpha subunit</protein> of the <protein>IL-2 receptor</protein> ) in <cell_type>T cells</cell_type> stimulated with phorbol ester .


These data demonstrate that DMDTC is a potent immunosuppressive compound in vitro .


Human 15-lipoxygenase gene promoter : analysis and identification of DNA binding sites for IL-13-induced regulatory factors in monocytes .

<dna>Human 15-lipoxygenase gene promoter</dna> : analysis and identification of <dna>DNA binding sites</dna> for <protein>IL-13-induced regulatory factors</protein> in <cell_type>monocytes</cell_type> .


In order to study the transcriptional control of 15-LO expression , we have cloned and sequenced the human 15-LO promoter region .

In order to study the transcriptional control of <protein>15-LO</protein> expression , we have cloned and sequenced the <dna>human 15-LO promoter region</dna> .


The 15-LO promoter is associated with a CpG island at the 5'-end of the gene , and sequence analysis reveals putative Sp1 and Ap2 binding site/s and absence of TATA or CAAT motifs .

The <protein>15-LO</protein> promoter is associated with a CpG island at the <dna>5'-end</dna> of the gene , and sequence analysis reveals putative <dna>Sp1 and Ap2 binding site/s</dna> and absence of <dna>TATA or CAAT motifs</dna> .


Transcription is initiated at one major site .


Using deletion constructs , we have defined an active promoter region of 1056 bp .

Using <dna>deletion constructs</dna> , we have defined an <dna>active promoter region</dna> of 1056 bp .


Gel-shift assays revealed that transcriptional factor ( s ) induced only in response to IL-13 treatment of human peripheral blood monocytes bind to the 15-LO promoter DNA .

Gel-shift assays revealed that <protein>transcriptional factor</protein> ( s ) induced only in response to <protein>IL-13</protein> treatment of <cell_type>human peripheral blood monocytes</cell_type> bind to the <dna>15-LO promoter DNA</dna> .


Two regions , DP1 ( -140 to -92 bp ) and DP2 ( -353 to -304 bp ) of the promoter were essential for transcription in HeLa cells and human peripheral monocytes .

Two regions , <dna>DP1</dna> ( <dna>-140 to -92 bp</dna> ) and <dna>DP2</dna> ( <dna>-353 to -304 bp</dna> ) of the promoter were essential for transcription in <cell_line>HeLa cells</cell_line> and <cell_type>human peripheral monocytes</cell_type> .


Hela nuclear extracts contained a specific nuclear factor ( s ) binding to 15-LO promoter DNA which are distinct from those derived from IL-13-treated human peripheral monocyte nuclear extracts .

Hela nuclear extracts contained a <protein>specific nuclear factor ( s )</protein> binding to <dna>15-LO promoter DNA</dna> which are distinct from those derived from <cell_line>IL-13-treated human peripheral monocyte</cell_line> nuclear extracts .


In addition , fluorescent in situ hybridization ( FISH ) results refined the previous localization of 15-LO to human chromosome 17p13.3 .

In addition , fluorescent in situ hybridization ( FISH ) results refined the previous localization of <protein>15-LO</protein> to <dna>human chromosome 17p13.3</dna> .


Ex vivo activation of tumor-draining lymph node T cells reverses defects in signal transduction molecules .

Ex vivo activation of <cell_type>tumor-draining lymph node T cells</cell_type> reverses defects in <protein>signal transduction molecules</protein> .


The adoptive transfer of tumor-draining lymph node ( LN ) T cells activated ex vivo with anti-CD3 and interleukin 2 ( IL-2 ) mediates the regression of the poorly immunogenic murine melanoma D5 .

The adoptive transfer of tumor-draining lymph node ( LN ) <cell_type>T cells</cell_type> activated ex vivo with <protein>anti-CD3</protein> and <protein>interleukin 2</protein> ( <protein>IL-2</protein> ) mediates the regression of the poorly immunogenic murine melanoma D5 .


The efficacy of the activated LN cells is augmented when the sensitizing tumor is a genetically modified variant ( designated D5G6 ) that secretes granulocyte/macrophage-colony-stimulating factor .

The efficacy of the <cell_type>activated LN cells</cell_type> is augmented when the sensitizing tumor is a <cell_line>genetically modified variant</cell_line> ( designated <cell_line>D5G6</cell_line> ) that secretes <protein>granulocyte/macrophage-colony-stimulating factor</protein> .


In contrast to anti-CD3/IL-2-activated LN cells , adoptive transfer of freshly isolated tumor-draining LN T cells has no therapeutic activity .

In contrast to <cell_line>anti-CD3/IL-2-activated LN cells</cell_line> , adoptive transfer of freshly isolated <cell_type>tumor-draining LN T cells</cell_type> has no therapeutic activity .


To determine whether the acquisition of antitumor function during ex vivo activation is associated with modifications in signal transduction capacity , the protein tyrosine kinases p56lck and p59fyn and proteins of the NF-kappaB family were analyzed in tumor-draining LN T cells .

To determine whether the acquisition of antitumor function during ex vivo activation is associated with modifications in signal transduction capacity , the protein <protein>tyrosine kinases</protein> <protein>p56lck</protein> and <protein>p59fyn</protein> and proteins of the <protein>NF-kappaB family</protein> were analyzed in <cell_type>tumor-draining LN T cells</cell_type> .


The levels of p56lck and p59fyn were lower in tumor-draining than in normal LN T cells and production of tyrosine-phosphorylated substrates was markedly depressed following anti-CD3 stimulation .

The levels of <protein>p56lck</protein> and <protein>p59fyn</protein> were lower in tumor-draining than in <cell_type>normal LN T cells</cell_type> and production of <protein>tyrosine-phosphorylated substrates</protein> was markedly depressed following <protein>anti-CD3</protein> stimulation .


After 5-day anti-CD3 /IL-2 activation , levels of p56lck and p59fyn and protein tyrosine kinase activity increased .

After 5-day <protein>anti-CD3</protein> <protein>/IL-2</protein> activation , levels of <protein>p56lck</protein> and <protein>p59fyn</protein> and protein tyrosine kinase activity increased .


Interestingly , the levels of p56lck , p59fyn , and tyrosine kinase activity were higher in activated T cells derived from LN that drained D5G6 than they were in those from D5 tumors .

Interestingly , the levels of <protein>p56lck</protein> , <protein>p59fyn</protein> , and tyrosine kinase activity were higher in <cell_type>activated T cells</cell_type> derived from LN that drained D5G6 than they were in those from D5 tumors .


In contrast , the cytoplasmic levels of c-Rel and Rel A were normal in freshly isolated tumor-draining LN , as was nuclear kappaB DNA-binding activity induced by anti-CD3 mAb or phorbol myristate acetate .

In contrast , the cytoplasmic levels of <protein>c-Rel</protein> and <protein>Rel A</protein> were normal in freshly isolated tumor-draining LN , as was nuclear <protein>kappaB</protein> DNA-binding activity induced by <protein>anti-CD3 mAb</protein> or phorbol myristate acetate .


Stimulation of activated LN cells with D5 tumor cells induced the nuclear translocation of NF-kappaB .

Stimulation of <cell_type>activated LN cells</cell_type> with <cell_type>D5 tumor cells</cell_type> induced the nuclear translocation of <protein>NF-kappaB</protein> .


These findings indicate that the recovery of proteins mediating signal transduction through the T cell receptor/CD3 complex in LN T cells activated ex vivo was associated with the acquisition of antitumor function .

These findings indicate that the recovery of proteins mediating signal transduction through the <protein>T cell receptor/CD3 complex</protein> in <cell_type>LN T cells</cell_type> activated ex vivo was associated with the acquisition of antitumor function .


Suppression of human anti-porcine T-cell immune responses by major histocompatibility complex class II transactivator constructs lacking the amino terminal domain .

Suppression of human anti-porcine T-cell immune responses by <dna>major histocompatibility complex class II transactivator constructs</dna> lacking the amino terminal domain .


BACKGROUND : The class II transactivator ( CIITA ) is a bi- or multifunctional domain protein that acts as a transcriptional activator and plays a critical role in the expression of MHC class II genes .

BACKGROUND : The <protein>class II transactivator</protein> ( <protein>CIITA</protein> ) is a <protein>bi- or multifunctional domain protein</protein> that acts as a <protein>transcriptional activator</protein> and plays a critical role in the expression of <dna>MHC class II genes</dna> .


We have previously demonstrated that a mutated form of the human CIITA gene , coding for a protein lacking the amino terminal 151 amino acids , acts as a potent dominant-negative suppressor of HLA class II expression .

We have previously demonstrated that a mutated form of the <dna>human CIITA gene</dna> , coding for a protein lacking the <protein>amino terminal 151 amino acids</protein> , acts as a potent dominant-negative suppressor of <protein>HLA class II</protein> expression .


Porcine MHC class II antigens are potent stimulators of direct T-cell recognition by human CD4+ T cells and are , therefore , likely to play an important role in the rejection responses to transgenic pig donors in clinical xenotransplantation .

<protein>Porcine MHC class II antigens</protein> are potent stimulators of direct T-cell recognition by <cell_type>human CD4+ T cells</cell_type> and are , therefore , likely to play an important role in the rejection responses to transgenic pig donors in clinical xenotransplantation .


We were , therefore , interested in examining mutated CIITA constructs for their effect on porcine MHC class II expression .

We were , therefore , interested in examining mutated <dna>CIITA constructs</dna> for their effect on porcine <protein>MHC class II</protein> expression .


METHODS : Stable transfectants of the porcine vascular endothelial cell line PIEC with mutated CIITA constructs were tested for SLA-DR and SLA-DQ induction by recombinant porcine interferon-gamma .

METHODS : Stable transfectants of the <cell_line>porcine vascular endothelial cell line</cell_line> <cell_line>PIEC</cell_line> with mutated <dna>CIITA constructs</dna> were tested for <protein>SLA-DR</protein> and <protein>SLA-DQ</protein> induction by <protein>recombinant porcine interferon-gamma</protein> .


Transient transfectants of the porcine B-cell line L23 with the mutated CIITA constructs were tested for the suppression of constitutive SLA-DR and SLA-DQ expression .

Transient transfectants of the <cell_line>porcine B-cell line L23</cell_line> with the mutated <dna>CIITA constructs</dna> were tested for the suppression of constitutive SLA-DR and SLA-DQ expression .


T-cell proliferation studies were performed using highly purified human CD4+ T cells .

T-cell proliferation studies were performed using highly purified <cell_type>human CD4+ T cells</cell_type> .


RESULTS : In preliminary studies , we demonstrated that transfection of the PIEC line with full-length human CIITA constructs resulted in strong expression of SLA-DR and SLA-DQ antigens , thus establishing the cross-species effectiveness of human CIITA in the pig .

RESULTS : In preliminary studies , we demonstrated that transfection of the <cell_line>PIEC line</cell_line> with full-length human <dna>CIITA constructs</dna> resulted in strong expression of <protein>SLA-DR</protein> and <protein>SLA-DQ</protein> antigens , thus establishing the cross-species effectiveness of <protein>human CIITA</protein> in the pig .


The mutated human CIITA constructs were , therefore , tested in the pig .

The <dna>mutated human CIITA constructs</dna> were , therefore , tested in the pig .


PIEC clones stably transfected with one of these constructs showed up to 99 % suppression of SLA-DR and SLA-DQ antigen induction and marked suppression of SLA-DRA mRNA induction .

<cell_line>PIEC clones</cell_line> stably transfected with one of these constructs showed up to 99 % suppression of <protein>SLA-DR</protein> and <protein>SLA-DQ</protein> antigen induction and marked suppression of <rna>SLA-DRA mRNA</rna> induction .


Moreover , transient transfection of the porcine B-cell line L23 showed up to 90 % suppression of constitutive SLA-DR and SLA-DQ antigen expression in 5-8 days .

Moreover , transient transfection of the <cell_line>porcine B-cell line L23</cell_line> showed up to 90 % suppression of constitutive SLA-DR and SLA-DQ antigen expression in 5-8 days .


In functional studies , interferon-gamma-stimulated PIEC clones transfected with this mutated CIITA construct failed to stimulate purified human CD4+ T lymphocytes .

In functional studies , <cell_line>interferon-gamma-stimulated PIEC clones</cell_line> transfected with this <protein>mutated CIITA</protein> construct failed to stimulate purified <cell_type>human CD4+ T lymphocytes</cell_type> .


CONCLUSION : Mutated human CIITA constructs are potent suppressors of porcine MHC class II expression .

CONCLUSION : <dna>Mutated human CIITA constructs</dna> are potent suppressors of porcine <protein>MHC class II</protein> expression .


Macrophages in human atheroma contain PPARgamma : differentiation-dependent peroxisomal proliferator-activated receptor gamma ( PPARgamma ) expression and reduction of MMP-9 activity through PPARgamma activation in mononuclear phagocytes in vitro .

<cell_type>Macrophages</cell_type> in human atheroma contain <protein>PPARgamma</protein> : <protein>differentiation-dependent peroxisomal proliferator-activated receptor gamma</protein> ( <protein>PPARgamma</protein> ) expression and reduction of <protein>MMP-9</protein> activity through <protein>PPARgamma</protein> activation in mononuclear phagocytes in vitro .


Mononuclear phagocytes play an important role in atherosclerosis and its sequela plaque rupture in part by their secretion of matrix metalloproteinases ( MMPs ) , including MMP-9 .

<cell_type>Mononuclear phagocytes</cell_type> play an important role in atherosclerosis and its sequela plaque rupture in part by their secretion of <protein>matrix metalloproteinases</protein> ( <protein>MMPs</protein> ) , including <protein>MMP-9</protein> .


Peroxisomal proliferator-activated receptor gamma ( PPARgamma ) , a transcription factor in the nuclear receptor superfamily , regulates gene expression in response to various activators , including 15-deoxy-delta12 , 14-prostaglandin J2 and the antidiabetic agent troglitazone .

<protein>Peroxisomal proliferator-activated receptor gamma</protein> ( <protein>PPARgamma</protein> ) , a <protein>transcription factor</protein> in the nuclear receptor superfamily , regulates gene expression in response to various activators , including 15-deoxy-delta12 , 14-prostaglandin J2 and the antidiabetic agent troglitazone .


The role of PPARgamma in human atherosclerosis is unexplored .

The role of <protein>PPARgamma</protein> in human atherosclerosis is unexplored .


We report here that monocytes/macrophages in human atherosclerotic lesions ( n = 12 ) express immunostainable PPARgamma .

We report here that <cell_type>monocytes/macrophages</cell_type> in human atherosclerotic lesions ( n = 12 ) express <protein>immunostainable PPARgamma</protein> .


Normal artery specimens ( n = 6 ) reveal minimal immunoreactive PPARgamma .

Normal artery specimens ( n = 6 ) reveal <protein>minimal immunoreactive PPARgamma</protein> .


Human monocytes and monocyte-derived macrophages cultured for 6 days in 5 % human serum expressed PPARgamma mRNA and protein by reverse transcription-polymerase chain reaction and Western blotting , respectively .

<cell_type>Human monocytes</cell_type> and <cell_type>monocyte-derived macrophages</cell_type> cultured for 6 days in 5 % human serum expressed <rna>PPARgamma mRNA</rna> and protein by reverse transcription-polymerase chain reaction and Western blotting , respectively .


In addition , PPARgamma mRNA expression in U937 cells increased during phorbol 12-myristate 13 acetate-induced differentiation .

In addition , <rna>PPARgamma mRNA</rna> expression in <cell_line>U937 cells</cell_line> increased during phorbol 12-myristate 13 acetate-induced differentiation .


Stimulation of PPARgamma with troglitazone or 15-deoxy-delta12 , 14-prostaglandin J2 in human monocyte-derived macrophages inhibited MMP-9 gelatinolytic activity in a concentration-dependent fashion as revealed by zymography .

Stimulation of <protein>PPARgamma</protein> with troglitazone or 15-deoxy-delta12 , 14-prostaglandin J2 in human <cell_type>monocyte-derived macrophages</cell_type> inhibited <protein>MMP-9</protein> gelatinolytic activity in a concentration-dependent fashion as revealed by zymography .


This inhibition correlates with decreased MMP-9 secretion as determined by Western blotting .

This inhibition correlates with decreased <protein>MMP-9</protein> secretion as determined by Western blotting .


Thus , PPARgamma is present in macrophages in human atherosclerotic lesions and may regulate expression and activity of MMP-9 , an enzyme implicated in plaque rupture .

Thus , <protein>PPARgamma</protein> is present in <cell_type>macrophages</cell_type> in human atherosclerotic lesions and may regulate expression and activity of <protein>MMP-9</protein> , an enzyme implicated in plaque rupture .


PPARgamma is likely to be an important regulator of monocyte/macrophage function with relevance for human atherosclerotic disease .

<protein>PPARgamma</protein> is likely to be an important regulator of monocyte/macrophage function with relevance for human atherosclerotic disease .


Erythropoietin induces tyrosine phosphorylation of Jak2 , STAT5A , and STAT5B in primary cultured human erythroid precursors .

<protein>Erythropoietin</protein> induces tyrosine phosphorylation of <protein>Jak2</protein> , <protein>STAT5A</protein> , and <protein>STAT5B</protein> in <cell_type>primary cultured human erythroid precursors</cell_type> .


We examined signaling by erythropoietin in highly purified human colony forming unit-erythroid cells , generated in vitro from CD34 ( + ) cells .

We examined signaling by <protein>erythropoietin</protein> in <cell_type>highly purified human colony forming unit-erythroid cells</cell_type> , generated in vitro from <cell_type>CD34 ( + ) cells</cell_type> .


We found that erythropoietin induces tyrosine phosphorylation of Jak2 , STAT5A , and STAT5B .

We found that <protein>erythropoietin</protein> induces tyrosine phosphorylation of <protein>Jak2</protein> , <protein>STAT5A</protein> , and <protein>STAT5B</protein> .


Tyrosine phosphorylation of Jak2 reaches a peak around 10 minutes after stimulation and is maximum at 5 U/mL of erythropoietin .

Tyrosine phosphorylation of <protein>Jak2</protein> reaches a peak around 10 minutes after stimulation and is maximum at 5 U/mL of <protein>erythropoietin</protein> .


Tyrosine phosphorylation of STAT5 is accompanied by the translocation of activated STAT5 to the nucleus as shown by electrophoretic mobility shift assay ( EMSA ) using 32Pi-labeled STAT5 binding site in the beta-casein promoter .

Tyrosine phosphorylation of <protein>STAT5</protein> is accompanied by the translocation of activated <protein>STAT5</protein> to the nucleus as shown by electrophoretic mobility shift assay ( EMSA ) using 32Pi-labeled <dna>STAT5 binding site</dna> in the <dna>beta-casein promoter</dna> .


Tyrosine phosphorylation STAT1 or STAT3 was not detected in human erythroid precursors after stimulation with erythropoietin .

Tyrosine phosphorylation <protein>STAT1</protein> or <protein>STAT3</protein> was not detected in <cell_type>human erythroid precursors</cell_type> after stimulation with <protein>erythropoietin</protein> .


Crkl , an SH2/SH3 adapter protein , becomes coimmunoprecipitated specifically with STAT5 from erythropoietin-stimulated erythroid cells ; although it was shown to become associated with c-Cbl in the studies using cell lines .

<protein>Crkl</protein> , an <protein>SH2/SH3 adapter protein</protein> , becomes coimmunoprecipitated specifically with <protein>STAT5</protein> from <cell_type>erythropoietin-stimulated erythroid cells</cell_type> ; although it was shown to become associated with <protein>c-Cbl</protein> in the studies using <cell_line>cell lines</cell_line> .


Thus , human erythroid precursors can be expanded in vitro in sufficient numbers and purity to allow its usage in signal transduction studies .

Thus , <cell_type>human erythroid precursors</cell_type> can be expanded in vitro in sufficient numbers and purity to allow its usage in signal transduction studies .


This report sets a basis for further studies on signaling in primary cultured human erythroid precursors , which in turn contribute to our better understanding in the differentiation processes of erythrocytes and their precursors .

This report sets a basis for further studies on signaling in <cell_type>primary cultured human erythroid precursors</cell_type> , which in turn contribute to our better understanding in the differentiation processes of <cell_type>erythrocytes</cell_type> and their precursors .


Oxidative stress suppresses transcription factor activities in stimulated lymphocytes .

Oxidative stress suppresses <protein>transcription factor</protein> activities in <cell_type>stimulated lymphocytes</cell_type> .


Effects of oxidative stress on stimulation-dependent signal transduction , leading to IL-2 expression , were studied .

Effects of oxidative stress on stimulation-dependent signal transduction , leading to <protein>IL-2</protein> expression , were studied .


Purified quiescent human blood T lymphocytes were subjected to : ( i ) acute exposure to hydrogen peroxide ; ( ii ) chronic exposure to hydrogen peroxide ; and ( iii ) acute exposure to ionizing radiation .

<cell_type>Purified quiescent human blood T lymphocytes</cell_type> were subjected to : ( i ) acute exposure to hydrogen peroxide ; ( ii ) chronic exposure to hydrogen peroxide ; and ( iii ) acute exposure to ionizing radiation .


The cells were then stimulated for 6 h .


DNA-binding activities ( determined by the electrophoretic mobility shift assay ) of three transcription factors : NFkappaB , AP-1 and NFAT , were abolished in the lymphocytes by all three modes of oxidative stress .

DNA-binding activities ( determined by the electrophoretic mobility shift assay ) of three <protein>transcription factors</protein> : <protein>NFkappaB</protein> , <protein>AP-1</protein> and <protein>NFAT</protein> , were abolished in the <cell_type>lymphocytes</cell_type> by all three modes of oxidative stress .


The lymphocytes exhibited lipid peroxidation only upon exposure to the lowest level of hydrogen peroxide used ( 20 microM ) .

The <cell_type>lymphocytes</cell_type> exhibited lipid peroxidation only upon exposure to the lowest level of hydrogen peroxide used ( 20 microM ) .


All three modes of oxidative stress induced catalase activity in the lymphocytes .

All three modes of oxidative stress induced <protein>catalase</protein> activity in the <cell_type>lymphocytes</cell_type> .


The only exception was hydrogen peroxide at 20 microM , which did not induce catalase activity .

The only exception was hydrogen peroxide at 20 microM , which did not induce <protein>catalase</protein> activity .


We conclude that : ( i ) suppression of specific transcription factor functions can potentially serve as a marker of exposure to oxidative stress and its effects on human lymphocytes ; ( ii ) lipid peroxidation is only detectable in human lymphocytes upon exposure to weak oxidative stress which does not induce catalase activity ; ( iii ) therefore , transcription factor DNA-binding activities are more sensitive to oxidative stress than lipid peroxidation .

We conclude that : ( i ) suppression of specific <protein>transcription factor</protein> functions can potentially serve as a marker of exposure to oxidative stress and its effects on <cell_type>human lymphocytes</cell_type> ; ( ii ) lipid peroxidation is only detectable in <cell_type>human lymphocytes</cell_type> upon exposure to weak oxidative stress which does not induce <protein>catalase</protein> activity ; ( iii ) therefore , <protein>transcription factor</protein> DNA-binding activities are more sensitive to oxidative stress than lipid peroxidation .


IL-4 -dependent regulation of TGF-alpha and TGF-beta1 expression in human eosinophils .

<protein>IL-4</protein> -dependent regulation of TGF-alpha and TGF-beta1 expression in <cell_type>human eosinophils</cell_type> .


TGFs play important roles in wound healing and carcinogenesis .

<protein>TGFs</protein> play important roles in wound healing and carcinogenesis .


We have previously demonstrated that eosinophils infiltrating into different pathologic processes elaborate TGF-alpha and TGF-beta1 .

We have previously demonstrated that <cell_type>eosinophils</cell_type> infiltrating into different pathologic processes elaborate <protein>TGF-alpha</protein> and <protein>TGF-beta1</protein> .


Eosinophils infiltrating hamster cutaneous wounds were found to express TGFs sequentially .

<cell_type>Eosinophils</cell_type> infiltrating hamster cutaneous wounds were found to express <protein>TGFs</protein> sequentially .


In this study , we examined the biologic mediators that may regulate the expression of TGF-alpha and -beta1 by eosinophils .

In this study , we examined the biologic mediators that may regulate the expression of <protein>TGF-alpha and -beta1</protein> by <cell_type>eosinophils</cell_type> .


Eosinophils were isolated from the peripheral blood of healthy donors and cultured in the absence or presence of IL-3 , IL-4 , and IL-5 .

<cell_type>Eosinophils</cell_type> were isolated from the peripheral blood of healthy donors and cultured in the absence or presence of <protein>IL-3</protein> , <protein>IL-4</protein> , and <protein>IL-5</protein> .


Cells were analyzed by in situ hybridization and immunohistochemistry .


Supernatants from these cultures were assayed for secreted TGF-alpha and TGF-beta1 using TGF -specific ELISAs .

Supernatants from these cultures were assayed for secreted <protein>TGF-alpha</protein> and <protein>TGF-beta1</protein> using <protein>TGF</protein> -specific ELISAs .


IL-3 , IL-4 , and IL-5 independently up-regulated TGF-beta1 mRNA and product expression by eosinophils in all donors .

<protein>IL-3</protein> , <protein>IL-4</protein> , and <protein>IL-5</protein> independently up-regulated <rna>TGF-beta1 mRNA</rna> and product expression by <cell_type>eosinophils</cell_type> in all donors .


Interestingly , TGF-alpha production by eosinophils was up-regulated by IL-3 and IL-5 but was down-regulated by IL-4 .

Interestingly , <protein>TGF-alpha</protein> production by <cell_type>eosinophils</cell_type> was up-regulated by <protein>IL-3</protein> and <protein>IL-5</protein> but was down-regulated by <protein>IL-4</protein> .


Consistent with the ability of IL-4 to regulate eosinophil responses , IL-4 signaling molecules are present in human eosinophils .

Consistent with the ability of <protein>IL-4</protein> to regulate eosinophil responses , <protein>IL-4 signaling molecules</protein> are present in <cell_type>human eosinophils</cell_type> .


The observation that IL-4 can differentially regulate the expression of TGF-alpha and TGF-beta1 suggests that IL-4 may serve as a physiologic molecular switch of TGF expression by the infiltrating eosinophils in wound healing and carcinogenesis .

The observation that <protein>IL-4</protein> can differentially regulate the expression of <protein>TGF-alpha</protein> and <protein>TGF-beta1</protein> suggests that <protein>IL-4</protein> may serve as a physiologic molecular switch of <protein>TGF</protein> expression by the infiltrating <cell_type>eosinophils</cell_type> in wound healing and carcinogenesis .


MLL and CALM are fused to AF10 in morphologically distinct subsets of acute leukemia with translocation t ( 10 ; 11 ) : both rearrangements are associated with a poor prognosis .

MLL and <dna>CALM</dna> are fused to <dna>AF10</dna> in morphologically distinct subsets of acute leukemia with <dna>translocation t ( 10 ; 11 )</dna> : both rearrangements are associated with a poor prognosis .


The translocation t ( 10 ; 11 ) ( p13 ; q14 ) has been observed in acute lymphoblastic leukemia ( ALL ) as well as acute myeloid leukemia ( AML ) .

The <dna>translocation t ( 10 ; 11 ) ( p13 ; q14 )</dna> has been observed in acute lymphoblastic leukemia ( ALL ) as well as acute myeloid leukemia ( AML ) .


A recent study showed a MLL/AF10 fusion in all cases of AML with t ( 10 ; 11 ) and various breakpoints on chromosome 11 ranging from q13 to q23 .

A recent study showed a <dna>MLL/AF10 fusion</dna> in all cases of AML with <dna>t ( 10 ; 11 )</dna> and various breakpoints on <dna>chromosome 11</dna> ranging from <dna>q13</dna> to <dna>q23</dna> .


We recently cloned CALM ( Clathrin Assembly Lymphoid Myeloid leukemia gene ) , the fusion partner of AF10 at 11q14 in the monocytic cell line U937 .

We recently cloned <dna>CALM</dna> ( <dna>Clathrin Assembly Lymphoid Myeloid leukemia gene</dna> ) , the fusion partner of <dna>AF10</dna> at <dna>11q14</dna> in the <cell_line>monocytic cell line U937</cell_line> .


To further define the role of these genes in acute leukemias , 10 cases ( 9 AML and 1 ALL ) with cytogenetically proven t ( 10 ; 11 ) ( p12-14 ; q13-21 ) and well-characterized morphology , immunophenotype , and clinical course were analyzed .

To further define the role of these genes in acute leukemias , 10 cases ( 9 AML and 1 ALL ) with cytogenetically proven <dna>t ( 10 ; 11 ) ( p12-14 ; q13-21 )</dna> and well-characterized morphology , immunophenotype , and clinical course were analyzed .


Interphase fluorescence in situ hybridization ( FISH ) was performed with 2 YACs flanking the CALM region , a YAC contig of the MLL region , and a YAC spanning the AF10 breakpoint .

Interphase fluorescence in situ hybridization ( FISH ) was performed with 2 <dna>YACs</dna> flanking the <dna>CALM region</dna> , a <dna>YAC contig</dna> of the <dna>MLL region</dna> , and a <dna>YAC</dna> spanning the <dna>AF10</dna> breakpoint .


Rearrangement of at least one of these genes was detected in all cases with balanced t ( 10 ; 11 ) .

Rearrangement of at least one of these genes was detected in all cases with balanced <dna>t ( 10 ; 11 )</dna> .


In 4 cases , including 3 AML with immature morphology ( 1 AML-M0 and 2 AML-M1 ) and 1 ALL , the signals of the CALM YACS were separated in interphase cells , indicating a translocation breakpoint within the CALM region .

In 4 cases , including 3 AML with immature morphology ( 1 AML-M0 and 2 AML-M1 ) and 1 ALL , the signals of the <dna>CALM YACS</dna> were separated in interphase cells , indicating a translocation breakpoint within the <dna>CALM region</dna> .


MLL was rearranged in 3 AML with myelomonocytic differentiation ( 2 AML-M2 and 1 AML-M5 ) , including 1 secondary AML .


In all 3 cases , a characteristic immunophenotype was identified ( CD4+ , CD13 - , CD33+ , CD65s+ ) .

In all 3 cases , a characteristic immunophenotype was identified ( <protein>CD4+</protein> , <protein>CD13</protein> - , CD33+ , CD65s+ ) .


AF-10 was involved in 5 of 6 evaluable cases , including 1 case without detectable CALM or MLL rearrangement .

AF-10 was involved in 5 of 6 evaluable cases , including 1 case without detectable <dna>CALM</dna> or MLL rearrangement .


In 2 complex translocations , none of the three genes was rearranged .


All cases had a remarkably poor prognosis , with a mean survival of 9.6 +/- 6.6 months .


For the 7 AML cases that were uniformly treated according to the AMLCG86/92 protocols , disease-free and overall survival was significantly worse than for the overall study group ( P = .03 and P = .01 , respectively ) .


We conclude that the t ( 10 ; 11 ) ( p13 ; q14 ) indicates CALM and MLL rearrangements in morphologically distinct subsets of acute leukemia and may be associated with a poor prognosis .

We conclude that the <dna>t ( 10 ; 11 ) ( p13 ; q14 )</dna> indicates <dna>CALM and MLL rearrangements</dna> in morphologically distinct subsets of acute leukemia and may be associated with a poor prognosis .


The role of E-proteins in B- and T-lymphocyte development .

The role of <protein>E-proteins</protein> in B- and T-lymphocyte development .


Department of lymphocytes from hematopoietic stem cells is controlled , in part , by the activity of transcriptional regulatory proteins .

Department of <cell_type>lymphocytes</cell_type> from <cell_type>hematopoietic stem cells</cell_type> is controlled , in part , by the activity of <protein>transcriptional regulatory proteins</protein> .


In particular , one class of helix-loop-helix proteins , termed E-proteins , have been implicated in the regulation of gene expression during B-cell development .

In particular , one class of <protein>helix-loop-helix proteins</protein> , termed <protein>E-proteins</protein> , have been implicated in the regulation of gene expression during B-cell development .


Recent analysis of gene-targeted mice has allowed a direct assessment of the functional roles of several E-protein family members in hematopoiesis .

Recent analysis of gene-targeted mice has allowed a direct assessment of the functional roles of <protein>several E-protein family members</protein> in hematopoiesis .


In this review we describe the defects in B- and T- lymphocyte development in mice carrying targeted mutations in the E-protein genes and discuss our current understanding of the role of these proteins in lymphoid development

In this review we describe the defects in B- and T- lymphocyte development in mice carrying targeted mutations in the <dna>E-protein genes</dna> and discuss our current understanding of the role of these proteins in lymphoid development


Epstein-Barr virus-transforming protein latent infection membrane protein 1 activates transcription factor NF-kappaB through a pathway that includes the NF-kappaB-inducing kinase and the IkappaB kinases IKKalpha and IKKbeta .

<protein>Epstein-Barr virus-transforming protein</protein> <protein>latent infection membrane protein 1</protein> activates <protein>transcription factor NF-kappaB</protein> through a pathway that includes the <protein>NF-kappaB-inducing kinase</protein> and the <protein>IkappaB kinases</protein> <protein>IKKalpha</protein> and <protein>IKKbeta</protein> .


The Epstein-Barr virus oncoprotein latent infection membrane protein 1 ( LMP1 ) is a constitutively aggregated pseudo-tumor necrosis factor receptor ( TNFR ) that activates transcription factor NF-kappaB through two sites in its C-terminal cytoplasmic domain .

The <protein>Epstein-Barr virus oncoprotein</protein> <protein>latent infection membrane protein 1</protein> ( <protein>LMP1</protein> ) is a constitutively aggregated <protein>pseudo-tumor necrosis factor receptor</protein> ( <protein>TNFR</protein> ) that activates <protein>transcription factor NF-kappaB</protein> through two sites in its <protein>C-terminal cytoplasmic domain</protein> .


One site is similar to activated TNFRII in associating with TNFR-associated factors TRAF1 and TRAF2 , and the second site is similar to TNFRI in associating with the TNFRI death domain interacting protein TRADD .

One site is similar to activated <protein>TNFRII</protein> in associating with <protein>TNFR-associated factors</protein> <protein>TRAF1</protein> and <protein>TRAF2</protein> , and the second site is similar to <protein>TNFRI</protein> in associating with the <protein>TNFRI death domain interacting protein</protein> <protein>TRADD</protein> .


TNFRI has been recently shown to activate NF-kappaB through association with TRADD , RIP , and TRAF2 ; activation of the NF-kappaB-inducing kinase ( NIK ) ; activation of the IkappaB alpha kinases ( IKKalpha and IKKbeta ) ; and phosphorylation of IkappaB alpha .

<protein>TNFRI</protein> has been recently shown to activate <protein>NF-kappaB</protein> through association with <protein>TRADD</protein> , <protein>RIP</protein> , and <protein>TRAF2</protein> ; activation of the <protein>NF-kappaB-inducing kinase</protein> ( <protein>NIK</protein> ) ; activation of the <protein>IkappaB alpha kinases</protein> ( <protein>IKKalpha</protein> and <protein>IKKbeta</protein> ) ; and phosphorylation of <protein>IkappaB alpha</protein> .


IkappaB alpha phosphorylation on Ser-32 and Ser-36 is followed by its degradation and NF-kappaB activation .

<protein>IkappaB alpha</protein> phosphorylation on Ser-32 and Ser-36 is followed by its degradation and <protein>NF-kappaB</protein> activation .