The M-CSF gene was expressed in 6 cases and the FMS and the EGR-1 genes were expressed in 2 of the latter cases .

The <dna>M-CSF gene</dna> was expressed in 6 cases and the <dna>FMS</dna> and the <dna>EGR-1 genes</dna> were expressed in 2 of the latter cases .


An autocrine mechanism of growth could be hypothesized only for the 2 patients whose cells expressed both the M-CSF and FMS genes .

An autocrine mechanism of growth could be hypothesized only for the 2 patients whose cells expressed both the <dna>M-CSF and FMS genes</dna> .


No germline changes or rearrangements were observed in any of the genes studied .


Thus , deregulation of genes encoding for certain hemopoietic growth factors or receptors does not seem to represent a major mechanism of MDS progression .

Thus , deregulation of genes encoding for certain <protein>hemopoietic growth factors</protein> or receptors does not seem to represent a major mechanism of MDS progression .


A novel human homeobox gene distantly related to proboscipedia is expressed in lymphoid and pancreatic tissues .

A novel <dna>human homeobox gene</dna> distantly related to <dna>proboscipedia</dna> is expressed in lymphoid and pancreatic tissues .


A novel human homeobox gene , HB9 , was isolated from a cDNA library prepared from in vitro stimulated human tonsil B lymphocytes and from a human genomic library .

A novel <dna>human homeobox gene</dna> , <dna>HB9</dna> , was isolated from a <dna>cDNA library</dna> prepared from in <cell_type>vitro stimulated human tonsil B lymphocytes</cell_type> and from a <dna>human genomic library</dna> .


The HB9 gene is composed of 3 exons spread over 6 kilobases of DNA .

The <dna>HB9 gene</dna> is composed of 3 <dna>exons</dna> spread over 6 kilobases of DNA .


An open reading frame of 1206 nucleotides is in frame with a diverged homeodomain .

An <dna>open reading frame</dna> of 1206 nucleotides is in frame with a <dna>diverged homeodomain</dna> .


The predicted HB9 protein has a molecular mass of 41 kilodaltons and is enriched for alanine , glycine , and leucine .

The predicted <protein>HB9 protein</protein> has a molecular mass of 41 kilodaltons and is enriched for alanine , glycine , and leucine .


The HB9 homeodomain is most similar to that of the Drosophila melanogaster homeobox gene proboscipedia .

The <dna>HB9 homeodomain</dna> is most similar to that of the <dna>Drosophila melanogaster homeobox gene</dna> <dna>proboscipedia</dna> .


Northern blot analysis of poly ( A ) RNA purified from the human B cell line RPMI 8226 and from activated T cells revealed a major mRNA transcript of 2.2 kilobases .

Northern blot analysis of <rna>poly ( A ) RNA</rna> purified from the <cell_line>human B cell line RPMI 8226</cell_line> and from <cell_type>activated T cells</cell_type> revealed a <rna>major mRNA transcript</rna> of 2.2 kilobases .


Similar analysis of poly ( A ) RNA from a variety of adult tissues demonstrated HB9 transcripts in pancreas , small intestine , and colon .

Similar analysis of <rna>poly ( A ) RNA</rna> from a variety of adult tissues demonstrated <dna>HB9</dna> transcripts in pancreas , small intestine , and colon .


Reverse transcriptase-polymerase chain reaction was used to examine HB9 RNA transcripts in hematopoietic cell lines .

Reverse transcriptase-polymerase chain reaction was used to examine <rna>HB9 RNA transcripts</rna> in <cell_line>hematopoietic cell lines</cell_line> .


HB9 RNA transcripts were most prevalent in several human B cell lines and K562 cells .

<rna>HB9 RNA transcripts</rna> were most prevalent in several <cell_line>human B cell lines</cell_line> and <cell_line>K562 cells</cell_line> .


In addition , transcripts were detected in RNA prepared from tonsil B cells and in situ hybridization studies localized them in the germinal center region of adult tonsil .

In addition , transcripts were detected in RNA prepared from <cell_type>tonsil B cells</cell_type> and in situ hybridization studies localized them in the germinal center region of adult tonsil .


These findings suggest the involvement of HB9 in regulating gene transcription in lymphoid and pancreatic tissues .

These findings suggest the involvement of <dna>HB9</dna> in regulating gene transcription in lymphoid and pancreatic tissues .


Human immunodeficiency virus type 1 Tat upregulates interleukin-2 secretion in activated T cells .

<protein>Human immunodeficiency virus type 1 Tat</protein> upregulates <protein>interleukin-2</protein> secretion in <cell_type>activated T cells</cell_type> .


Dysregulation of cytokines secreted by T cells may play an important role in the pathogenesis of AIDS .

Dysregulation of <protein>cytokines</protein> secreted by <cell_type>T cells</cell_type> may play an important role in the pathogenesis of AIDS .


To investigate the effects of human immunodeficiency virus type 1 ( HIV-1 ) Tat on interleukin-2 ( IL-2 ) expression , we used IL-2 promoter-chloramphenicol acetyltransferase constructs and IL-2-secreting Jurkat T cells as a model system .

To investigate the effects of <protein>human immunodeficiency virus type 1 ( HIV-1 ) Tat</protein> on <protein>interleukin-2</protein> ( <protein>IL-2</protein> ) expression , we used <dna>IL-2 promoter-chloramphenicol acetyltransferase constructs</dna> and <cell_line>IL-2-secreting Jurkat T cells</cell_line> as a model system .


Transient expression of HIV-1 Tat induced a five- to eightfold increase in IL-2 promoter activity in Jurkat T cells stimulated with phytohemagglutinin and phorbol myristate acetate .

Transient expression of <protein>HIV-1 Tat</protein> induced a five- to eightfold increase in <dna>IL-2 promoter</dna> activity in <cell_line>Jurkat T cells</cell_line> stimulated with <protein>phytohemagglutinin</protein> and phorbol myristate acetate .


IL-2 secretion was increased more than twofold in both Jurkat T cells and primary T cells stimulated by extracellular HIV-1 Tat protein .

<protein>IL-2</protein> secretion was increased more than twofold in both <cell_line>Jurkat T cells</cell_line> and <cell_line>primary T cells</cell_line> stimulated by <protein>extracellular HIV-1 Tat protein</protein> .


Analysis of mRNA suggested that Tat exerts its effect on IL-2 primarily at the transcriptional level .

Analysis of <rna>mRNA</rna> suggested that Tat exerts its effect on <protein>IL-2</protein> primarily at the transcriptional level .


The NF-kappa B site at positions -206 to -195 of the IL-2 promoter was required but not sufficient for the Tat effect .

The <dna>NF-kappa B site</dna> at positions <dna>-206 to -195</dna> of the <dna>IL-2 promoter</dna> was required but not sufficient for the <protein>Tat</protein> effect .


The Tat -mediated increase in IL-2 promoter activity could selectively be blocked by antisense tat or-unlike the analogous effect of human T-cell lymphotropic virus type 1 Tax -by cyclosporin A .

The <protein>Tat</protein> -mediated increase in <dna>IL-2 promoter</dna> activity could selectively be blocked by antisense tat or-unlike the analogous effect of <protein>human T-cell lymphotropic virus type 1 Tax</protein> -by cyclosporin A .


The observed increase in IL-2 levels might facilitate virus spread from or to T cells .

The observed increase in <protein>IL-2</protein> levels might facilitate virus spread from or to <cell_type>T cells</cell_type> .


Furthermore , it might contribute to the hypergammaglobulinemia or , together with other cytokines found to be dysregulated , the T-helper cell dysfunctions observed in AIDS patients .

Furthermore , it might contribute to the hypergammaglobulinemia or , together with other <protein>cytokines</protein> found to be dysregulated , the T-helper cell dysfunctions observed in AIDS patients .


Activation of nuclear factor kappa B in human lymphoblastoid cells by low-dose ionizing radiation .

Activation of <protein>nuclear factor kappa B</protein> in <cell_type>human lymphoblastoid cells</cell_type> by low-dose ionizing radiation .


Nuclear factor kappa B ( NF-kappa B ) is a pleiotropic transcription factor which is involved in the transcriptional regulation of several specific genes .

<protein>Nuclear factor kappa B</protein> ( <protein>NF-kappa B</protein> ) is a <protein>pleiotropic transcription factor</protein> which is involved in the transcriptional regulation of several specific genes .


Recent reports demonstrated that ionizing radiation in the dose range of 2-50 Gy results in expression of NF-kappa B in human KG-1 myeloid leukemia cells and human B-lymphocyte precursor cells ; the precise mechanism involved and the significance are not yet known .

Recent reports demonstrated that ionizing radiation in the dose range of 2-50 Gy results in expression of <protein>NF-kappa B</protein> in <cell_line>human KG-1 myeloid leukemia cells</cell_line> and <cell_type>human B-lymphocyte precursor cells</cell_type> ; the precise mechanism involved and the significance are not yet known .


The present report demonstrates that even lower doses of ionizing radiation , 0.25-2.0 Gy , are capable of inducing expression of NF-kappa B in EBV-transformed 244B human lymphoblastoid cells .

The present report demonstrates that even lower doses of ionizing radiation , 0.25-2.0 Gy , are capable of inducing expression of <protein>NF-kappa B</protein> in <cell_line>EBV-transformed 244B human lymphoblastoid cells</cell_line> .


These results are in a dose range where the viability of the cells remains very high .


After exposure to 137Cs gamma rays at a dose rate of 1.17 Gy/min , a maximum in expression of NF-kappa B was seen at 8 h after a 0.5-Gy exposure .

After exposure to 137Cs gamma rays at a dose rate of 1.17 Gy/min , a maximum in expression of <protein>NF-kappa B</protein> was seen at 8 h after a 0.5-Gy exposure .


Time-course studies revealed a biphasic time-dependent expression after 0.5- , 1- and 2-Gy exposures .


However , for each time examined , the expression of NF-kappa B was maximum after the 0.5-Gy exposure .

However , for each time examined , the expression of <protein>NF-kappa B</protein> was maximum after the 0.5-Gy exposure .


The expression of the p50 and p65 NF-kappa B subunits was also shown to be regulated differentially after exposures to 1.0 and 2.0 Gy .

The expression of the <protein>p50 and p65 NF-kappa B subunits</protein> was also shown to be regulated differentially after exposures to 1.0 and 2.0 Gy .


Alternative splicing of RNA transcripts encoded by the murine p105 NF-kappa B gene generates I kappa B gamma isoforms with different inhibitory activities .

Alternative splicing of <rna>RNA transcripts</rna> encoded by the <dna>murine p105 NF-kappa B gene</dna> generates <protein>I kappa B gamma isoforms</protein> with different inhibitory activities .


The gene encoding the 105-kDa protein ( p105 ) precursor of the p50 subunit of transcription factor NF-kappa B also encodes a p70 I kappa B protein , I kappa B gamma , which is identical to the C-terminal 607 amino acids of p105 .

The gene encoding the <protein>105-kDa protein ( p105 ) precursor</protein> of the <protein>p50</protein> subunit of transcription factor <protein>NF-kappa B</protein> also encodes a <protein>p70 I kappa B protein</protein> , <protein>I kappa B gamma</protein> , which is identical to the <protein>C-terminal 607 amino acids</protein> of <protein>p105</protein> .


Here we show that alternative RNA splicing generates I kappa B gamma isoforms with properties different from those of p70 .

Here we show that alternative RNA splicing generates <protein>I kappa B gamma isoforms</protein> with properties different from those of <protein>p70</protein> .


One 63-kDa isoform , termed I kappa B gamma-1 , which lacks 59 amino acids C-terminal to ankyrin repeat 7 , has a novel 35-amino acid C terminus encoded by an alternative reading frame of the p105 gene .

One <protein>63-kDa isoform</protein> , termed <protein>I kappa B gamma-1</protein> , which lacks 59 amino acids C-terminal to <protein>ankyrin repeat 7</protein> , has a novel <protein>35-amino acid C terminus</protein> encoded by an <dna>alternative reading frame</dna> of the <dna>p105 gene</dna> .


A 55-kDa isoform , I kappa B gamma-2 , lacks the 190 C-terminal amino acids of p70 I kappa B gamma .

A <protein>55-kDa isoform</protein> , <protein>I kappa B gamma-2</protein> , lacks the <protein>190 C-terminal amino acids</protein> of <protein>p70</protein> <protein>I kappa B gamma</protein> .


In contrast to p70 I kappa B gamma , which is a cytoplasmic protein , I kappa B gamma-1 is found in both the cytoplasm and nucleus , whereas I kappa B gamma-2 is predominantly nuclear .

In contrast to <protein>p70</protein> <protein>I kappa B gamma</protein> , which is a <protein>cytoplasmic protein</protein> , <protein>I kappa B gamma-1</protein> is found in both the cytoplasm and nucleus , whereas <protein>I kappa B gamma-2</protein> is predominantly nuclear .


The I kappa B gamma isoforms also display differences in specificity and affinity for Rel/NF-kappa B proteins .

The <protein>I kappa B gamma isoforms</protein> also display differences in specificity and affinity for <protein>Rel/NF-kappa B proteins</protein> .


While p70 I kappa B gamma inhibits p50- , p65- , and c-Rel-mediated transactivation and/or DNA binding , both I kappa B gamma-1 and I kappa B gamma-2 are specific for p50 and have different affinities for this subunit .

While <protein>p70</protein> <protein>I kappa B gamma</protein> inhibits p50- , p65- , and c-Rel-mediated transactivation and/or DNA binding , both <protein>I kappa B gamma-1</protein> and <protein>I kappa B gamma-2</protein> are specific for <protein>p50</protein> and have different affinities for this subunit .


The absence in I kappa B gamma-1 and I kappa B gamma-2 of a protein kinase A site whose phosphorylation modulates p70 I kappa B gamma inhibitory activity suggests that alternative RNA splicing may be used to generate I kappa B gamma isoforms that respond differently to intracellular signals .

The absence in <protein>I kappa B gamma-1</protein> and <protein>I kappa B gamma-2</protein> of a <dna>protein kinase A site</dna> whose phosphorylation modulates <protein>p70</protein> <protein>I kappa B gamma</protein> inhibitory activity suggests that alternative RNA splicing may be used to generate <protein>I kappa B gamma isoforms</protein> that respond differently to intracellular signals .


Structure and expression of the human GATA3 gene .

Structure and expression of the <dna>human GATA3 gene</dna> .


GATA3 , a member of the GATA family that is abundantly expressed in the T-lymphocyte lineage , is thought to participate in T-cell receptor gene activation through binding to enhancers .

<protein>GATA3</protein> , a member of the <protein>GATA family</protein> that is abundantly expressed in the <cell_type>T-lymphocyte lineage</cell_type> , is thought to participate in <protein>T-cell receptor</protein> gene activation through binding to <dna>enhancers</dna> .


To understand GATA3 gene regulation , we cloned the human gene and the 5 ' end of the mouse GATA3 gene .

To understand <dna>GATA3 gene</dna> regulation , we cloned the <dna>human gene</dna> and the <dna>5 ' end</dna> of the <dna>mouse GATA3 gene</dna> .


We show that the human GATA3 gene contains six exons distributed over 17 kb of DNA .

We show that the <dna>human GATA3 gene</dna> contains six <dna>exons</dna> distributed over 17 kb of DNA .


The two human GATA3 zinc fingers are encoded by two separate exons highly conserved with those of GATA1 , but no other structural homologies between these two genes can be found .

The two <protein>human GATA3 zinc fingers</protein> are encoded by two separate <dna>exons</dna> highly conserved with those of <protein>GATA1</protein> , but no other structural homologies between these two genes can be found .


The human and mouse GATA3 transcription units start at a major initiation site .

The <dna>human and mouse GATA3 transcription units</dna> start at a <dna>major initiation site</dna> .


The promoter sequence analysis of these two genes revealed that they are embedded within a CpG island and share structural features often found in the promoters of housekeeping genes .

The promoter sequence analysis of these two genes revealed that they are embedded within a <dna>CpG island</dna> and share structural features often found in the <dna>promoters</dna> of <dna>housekeeping genes</dna> .


Finally , we show that a DNA fragment containing the human GATA3 transcription unit , 3 kb upstream from the initiation site and 4 kb downstream from the polyadenylation site , displays T-cell specificity .

Finally , we show that a <dna>DNA fragment</dna> containing the <dna>human GATA3 transcription unit</dna> , <dna>3 kb upstream</dna> from the <dna>initiation site</dna> and <dna>4 kb downstream</dna> from the <dna>polyadenylation site</dna> , displays T-cell specificity .


Characterization of the human gene encoding LBR , an integral protein of the nuclear envelope inner membrane .

Characterization of the <dna>human gene</dna> encoding <protein>LBR</protein> , an <protein>integral protein</protein> of the nuclear envelope inner membrane .


We have characterized the human gene encoding LBR , an integral protein of the nuclear envelope inner membrane .

We have characterized the <dna>human gene</dna> encoding <protein>LBR</protein> , an <protein>integral protein</protein> of the nuclear envelope inner membrane .


Restriction mapping shows that the transcription unit spans approximately 35 kilobases .

Restriction mapping shows that the <dna>transcription unit</dna> spans approximately 35 kilobases .


A transcription start site is located approximately 4 kilobases 5 ' to the translation initiation codon , and an RNA splice of 3863 bases occurs in the 5'-untranslated region to generate mature HeLa cell mRNA .

A transcription start site is located approximately 4 kilobases 5 ' to the <dna>translation initiation codon</dna> , and an RNA splice of 3863 bases occurs in the <dna>5'-untranslated region</dna> to generate <rna>mature HeLa cell mRNA</rna> .


5 ' to the identified transcription start site are two CCAAT sequences and potential recognition sites for several transcription factors including Sp1 , AP-1 , AP-2 , and NF-kB .

5 ' to the identified <dna>transcription start site</dna> are two <dna>CCAAT sequences</dna> and <dna>potential recognition sites</dna> for several <protein>transcription factors</protein> including <protein>Sp1</protein> , <protein>AP-1</protein> , <protein>AP-2</protein> , and <protein>NF-kB</protein> .


There are 13 protein coding exons in the LBR gene .

There are 13 protein coding <dna>exons</dna> in the <dna>LBR gene</dna> .


LBR 's nucleoplasmic domain is encoded by exons 1-4 , and its hydrophobic domain , with eight putative transmembrane segments , is encoded by exons 5-13 .

<protein>LBR</protein> 's <protein>nucleoplasmic domain</protein> is encoded by <dna>exons 1-4</dna> , and its <protein>hydrophobic domain</protein> , with eight putative <protein>transmembrane segments</protein> , is encoded by <dna>exons 5-13</dna> .


The hydrophobic domain is homologous to three yeast polypeptides , suggesting that this higher eukaryotic gene could have evolved from recombination between a gene that encoded a soluble nuclear protein and a membrane protein gene similar to those in yeast .

The <protein>hydrophobic domain</protein> is homologous to three yeast polypeptides , suggesting that this <dna>higher eukaryotic gene</dna> could have evolved from recombination between a gene that encoded a <protein>soluble nuclear protein</protein> and a <dna>membrane protein gene</dna> similar to those in yeast .


These results are the first to demonstrate the structural organization of a vertebrate gene encoding an integral membrane protein of the nuclear envelope that may be a member of a family of polypeptides conserved in evolution .

These results are the first to demonstrate the structural organization of a <dna>vertebrate gene</dna> encoding an <protein>integral membrane protein</protein> of the nuclear envelope that may be a member of a family of polypeptides conserved in evolution .


Retinoic acid-induced expression of CD38 antigen in myeloid cells is mediated through retinoic acid receptor-alpha .

Retinoic acid-induced expression of <protein>CD38 antigen</protein> in <cell_type>myeloid cells</cell_type> is mediated through <protein>retinoic acid receptor-alpha</protein> .


CD38 is a leukocyte differentiation antigen that has been thought to be a phenotypic marker of different subpopulations of T- and B- lymphocytes .

<protein>CD38</protein> is a <protein>leukocyte differentiation antigen</protein> that has been thought to be a phenotypic marker of different subpopulations of <cell_type>T- and B- lymphocytes</cell_type> .


In myeloid cells , CD38 is expressed during early stages of differentiation .

In <cell_type>myeloid cells</cell_type> , <protein>CD38</protein> is expressed during early stages of differentiation .


Virtually no information is available on regulation and functions of CD38 .

Virtually no information is available on regulation and functions of <protein>CD38</protein> .


Recently we reported that all-trans-retinoic acid ( ATRA ) is a potent and highly specific inducer of CD38 expression in human promyelocytic leukemia cells .

Recently we reported that all-trans-retinoic acid ( ATRA ) is a potent and highly specific inducer of <protein>CD38</protein> expression in <cell_type>human promyelocytic leukemia cells</cell_type> .


Here we report that ATRA-induced expression of CD38 antigen in myeloid cells is mediated through retinoic acid-alpha receptor ( RAR alpha ) .

Here we report that ATRA-induced expression of <protein>CD38 antigen</protein> in <cell_type>myeloid cells</cell_type> is mediated through <protein>retinoic acid-alpha receptor</protein> ( <protein>RAR alpha</protein> ) .


ATRA failed to induce CD38 expression in a mutant subclone of the HL-60 myeloid leukemia cell line ( designated HL-60R ) that is relatively resistant to ATRA-induced granulocytic differentiation .

ATRA failed to induce <protein>CD38</protein> expression in a <cell_line>mutant subclone</cell_line> of the <cell_line>HL-60 myeloid leukemia cell line</cell_line> ( designated <cell_line>HL-60R</cell_line> ) that is relatively resistant to ATRA-induced granulocytic differentiation .


Retroviral vector-mediated transduction of RA receptor ( RAR alpha ) into this HL-60R subclone completely restored the sensitivity of these cells to ATRA in terms of their ability to express CD38 .

Retroviral vector-mediated transduction of <protein>RA receptor</protein> ( <protein>RAR alpha</protein> ) into this <cell_line>HL-60R subclone</cell_line> completely restored the sensitivity of these cells to ATRA in terms of their ability to express <protein>CD38</protein> .


In contrast , CD38 expression was not inducible by ATRA in HL-60R cells , transfected with a functional RAR beta , RAR gamma , or RXR alpha receptor .

In contrast , <protein>CD38</protein> expression was not inducible by ATRA in <cell_line>HL-60R cells</cell_line> , transfected with a functional <protein>RAR beta</protein> , <protein>RAR gamma</protein> , or <protein>RXR alpha receptor</protein> .


Induction of CD38 in acute promyelocytic and acute myeloblastic leukemia cells was independent of ATRA-induced cytodifferentiation .

Induction of <protein>CD38</protein> in <cell_type>acute promyelocytic and acute myeloblastic leukemia cells</cell_type> was independent of ATRA-induced cytodifferentiation .


Following culture with ATRA , increased CD38 protein levels were also observed in normal CD34+ bone marrow cells , but not on normal circulating granulocytes .

Following culture with ATRA , increased <protein>CD38</protein> protein levels were also observed in <cell_type>normal CD34+ bone marrow cells</cell_type> , but not on <cell_type>normal circulating granulocytes</cell_type> .


From these results , we conclude that CD38 is ATRA inducible in myeloid leukemia cells and normal CD34+ bone marrow cells .

From these results , we conclude that <protein>CD38</protein> is ATRA inducible in <cell_type>myeloid leukemia cells</cell_type> and <cell_type>normal CD34+ bone marrow cells</cell_type> .


This effect is independent of differentiation and is mediated by RAR alpha in HL-60 cells , suggesting a similar role for RAR alpha in CD38 expression in other hematopoietic cells .

This effect is independent of differentiation and is mediated by <protein>RAR alpha</protein> in <cell_type>HL-60 cells</cell_type> , suggesting a similar role for <protein>RAR alpha</protein> in <protein>CD38</protein> expression in other <cell_type>hematopoietic cells</cell_type> .


Some antioxidants inhibit , in a co-ordinate fashion , the production of tumor necrosis factor-alpha , IL-beta , and IL-6 by human peripheral blood mononuclear cells .

Some antioxidants inhibit , in a co-ordinate fashion , the production of <protein>tumor necrosis factor-alpha</protein> , <protein>IL-beta</protein> , and <protein>IL-6</protein> by <cell_type>human peripheral blood mononuclear cells</cell_type> .


Some antioxidants , including butylated hydroxyanisole ( BHA ) , tetrahydropapaveroline ( THP ) , nordihydroguiauretic acid , and 10 , 11-dihydroxyaporphine ( DHA ) , were found to be potent inhibitors of the production of tumor necrosis factor ( TNF ) -alpha , IL-1 beta , and IL-6 by human peripheral blood mononuclear cells ( PBMC ) stimulated by lipopolysaccharide ( LPS ) ( IC50s in the low micromolar range ) .

Some antioxidants , including butylated hydroxyanisole ( BHA ) , tetrahydropapaveroline ( THP ) , nordihydroguiauretic acid , and 10 , 11-dihydroxyaporphine ( DHA ) , were found to be potent inhibitors of the production of <protein>tumor necrosis factor ( TNF ) -alpha</protein> , <protein>IL-1 beta</protein> , and <protein>IL-6</protein> by <cell_type>human peripheral blood mononuclear cells</cell_type> ( <cell_type>PBMC</cell_type> ) stimulated by lipopolysaccharide ( LPS ) ( IC50s in the low micromolar range ) .


Inhibition of cytokine production was gene selective and not due to general effects on protein synthesis .

Inhibition of <protein>cytokine</protein> production was gene selective and not due to general effects on protein synthesis .


Inhibition of cytokine production by PBMC was observed also when other inducers were used ( staphylococci , silica , zymosan ) .

Inhibition of <protein>cytokine</protein> production by <cell_type>PBMC</cell_type> was observed also when other inducers were used ( staphylococci , silica , zymosan ) .


Much higher concentrations of other antioxidants -- including ascorbic acid , trolox , alpha-tocopherol , butylated hydroxytoluene , and the 5-lipoxygenase inhibitor zileuton -- did not affect the production of these cytokines .

Much higher concentrations of other antioxidants -- including ascorbic acid , trolox , alpha-tocopherol , butylated hydroxytoluene , and the 5-lipoxygenase inhibitor zileuton -- did not affect the production of these <protein>cytokines</protein> .


The active compounds did not inhibit IL-1 -induced production of IL-6 in fibroblasts , showing the cell selectivity of the effect .

The active compounds did not inhibit <protein>IL-1</protein> -induced production of <protein>IL-6</protein> in <cell_type>fibroblasts</cell_type> , showing the cell selectivity of the effect .


Antioxidant-mediated inhibition of cytokine production was correlated with low levels of the corresponding messenger RNAs .

Antioxidant-mediated inhibition of <protein>cytokine</protein> production was correlated with low levels of the corresponding <rna>messenger RNAs</rna> .


Nuclear run-on experiments showed that THP inhibited transcription of the IL-1 beta gene .

Nuclear run-on experiments showed that THP inhibited transcription of the <dna>IL-1 beta gene</dna> .


THP decreased the concentration of the transcription factors NF-kappa B and AP-1 detected in nuclear extracts of PBMC cultured in the presence or absence of LPS .

THP decreased the concentration of the <protein>transcription factors</protein> <protein>NF-kappa B</protein> and <protein>AP-1</protein> detected in nuclear extracts of <cell_type>PBMC</cell_type> cultured in the presence or absence of LPS .


THP and DHA markedly decreased the levels of TNF-alpha and IL-1 beta in the circulation of mice following LPS injection .

THP and DHA markedly decreased the levels of <protein>TNF-alpha</protein> and <protein>IL-1 beta</protein> in the circulation of mice following LPS injection .


Thus antioxidants vary widely in potency as inhibitors of the activation of transcription factors and of the transcription of genes for pro-inflammatory cytokines .

Thus antioxidants vary widely in potency as inhibitors of the activation of <protein>transcription factors</protein> and of the transcription of <dna>genes</dna> for <protein>pro-inflammatory cytokines</protein> .


Coordinate inhibition of the transcription of genes for inflammatory cytokines could provide a strategy for therapy of diseases with inflammatory pathogenesis and for septic shock .

Coordinate inhibition of the transcription of genes for inflammatory <protein>cytokines</protein> could provide a strategy for therapy of diseases with inflammatory pathogenesis and for septic shock .


An interleukin-4-induced transcription factor : IL-4 Stat .

An <protein>interleukin-4-induced transcription factor</protein> : <protein>IL-4 Stat</protein> .


Interleukin-4 ( IL-4 ) is an immunomodulatory cytokine secreted by activated T lymphocytes , basophils , and mast cells .

<protein>Interleukin-4</protein> ( <protein>IL-4</protein> ) is an immunomodulatory cytokine secreted by activated <cell_type>T lymphocytes</cell_type> , <cell_type>basophils</cell_type> , and <cell_type>mast cells</cell_type> .


It plays an important role in modulating the balance of T helper ( Th ) cell subsets , favoring expansion of the Th2 lineage relative to Th1 .

It plays an important role in modulating the balance of <cell_type>T helper ( Th ) cell subsets</cell_type> , favoring expansion of the <cell_type>Th2 lineage</cell_type> relative to <cell_type>Th1</cell_type> .


Imbalance of these T lymphocyte subsets has been implicated in immunological diseases including allergy , inflammation , and autoimmune disease .

Imbalance of these <cell_type>T lymphocyte subsets</cell_type> has been implicated in immunological diseases including allergy , inflammation , and autoimmune disease .


IL-4 may mediate its biological effects , at least in part , by activating a tyrosine-phosphorylated DNA binding protein .

<protein>IL-4</protein> may mediate its biological effects , at least in part , by activating a <protein>tyrosine-phosphorylated DNA binding protein</protein> .


This protein has now been purified and its encoding gene cloned .


Examination of the primary amino acid sequence of this protein indicates that it is a member of the signal transducers and activators of transcription ( Stat ) family of DNA binding proteins , hereby designated IL-4 Stat .

Examination of the primary amino acid sequence of this protein indicates that it is a member of the <protein>signal transducers and activators of transcription ( Stat ) family</protein> of <protein>DNA binding proteins</protein> , hereby designated <protein>IL-4 Stat</protein> .


Study of the inhibitory activities of phosphotyrosine-containing peptides derived from the intracellular domain of the IL-4 receptor provided evidence for direct coupling of receptor and transcription factor during the IL-4 Stat activation cycle .

Study of the inhibitory activities of phosphotyrosine-containing peptides derived from the <protein>intracellular domain of the IL-4 receptor</protein> provided evidence for direct coupling of <protein>receptor</protein> and <protein>transcription factor</protein> during the <protein>IL-4 Stat</protein> activation cycle .


Such observations indicate that IL-4 Stat has the same functional domain for both receptor coupling and dimerization .

Such observations indicate that <protein>IL-4 Stat</protein> has the same <protein>functional domain</protein> for both receptor coupling and dimerization .


Evaluation of the respiratory epithelium of normals and individuals with cystic fibrosis for the presence of adenovirus E1a sequences relevant to the use of E1a- adenovirus vectors for gene therapy for the respiratory manifestations of cystic fibrosis .

Evaluation of the respiratory epithelium of normals and individuals with cystic fibrosis for the presence of <dna>adenovirus E1a sequences</dna> relevant to the use of E1a- adenovirus vectors for gene therapy for the respiratory manifestations of cystic fibrosis .


Lung disease associated with disorders such as cystic fibrosis ( CF ) may be amenable to somatic gene therapy in which there is delivery of the normal gene directly to the respiratory epithelium using E1a- adenovirus ( Ad ) type 2- or 5-based vectors .

Lung disease associated with disorders such as cystic fibrosis ( CF ) may be amenable to somatic gene therapy in which there is delivery of the <dna>normal gene</dna> directly to the respiratory epithelium using E1a- adenovirus ( Ad ) type 2- or 5-based vectors .


For safety reasons , the Ad vectors are rendered replication deficient by deletion of the E1a region .

For safety reasons , the Ad vectors are rendered replication deficient by deletion of the <dna>E1a region</dna> .


Because there is the theoretical possibility of an E1a- replication-deficient vector replicating as a result of recombination or complementation with Ad 2/5 E1a sequences present in the target cell , this study is directed toward evaluating respiratory epithelium of normals and individuals with CF for the presence of E1a sequences .

Because there is the theoretical possibility of an E1a- replication-deficient vector replicating as a result of recombination or complementation with <dna>Ad 2/5 E1a sequences</dna> present in the target cell , this study is directed toward evaluating respiratory epithelium of normals and individuals with CF for the presence of E1a sequences .


Using Ad 2/5 E1a-specific primers and the polymerase chain reaction to evaluate DNA recovered from freshly isolated nasal and bronchial epithelium recovered by brushing , E1a sequences were detected in respiratory epithelium of 19 of 91 normals ( 21 % ) .

Using <dna>Ad 2/5 E1a-specific primers</dna> and the polymerase chain reaction to evaluate DNA recovered from freshly isolated nasal and bronchial epithelium recovered by brushing , <dna>E1a sequences</dna> were detected in respiratory epithelium of 19 of 91 normals ( 21 % ) .


In the E1a-positive samples , the average of E1a copy number was 55 +/- 18/10 ( 3 ) recovered cells .


In CF individuals , 7 of 52 ( 13 % ) had detectable E1a sequences in the respiratory epithelium , with E1a copy number in the positive samples of 80 +/- 21/10 ( 3 ) recovered cells .

In CF individuals , 7 of 52 ( 13 % ) had detectable <dna>E1a sequences</dna> in the respiratory epithelium , with E1a copy number in the positive samples of 80 +/- 21/10 ( 3 ) recovered cells .


These results demonstrate that there are detectable Ad 2/5 E1a sequences in the respiratory epithelium of a small percentage of normals and individuals with CF .

These results demonstrate that there are detectable <dna>Ad 2/5 E1a sequences</dna> in the respiratory epithelium of a small percentage of normals and individuals with CF .


Because of the theoretical potential of such sequences supporting replication of E1a- Ad vectors , human gene therapy protocols for CF utilizing such vectors should consider evaluating study individuals for the presence of Ad 2/5 E1a sequences in the respiratory epithelium .

Because of the theoretical potential of such sequences supporting replication of E1a- Ad vectors , human gene therapy protocols for CF utilizing such vectors should consider evaluating study individuals for the presence of <dna>Ad 2/5 E1a sequences</dna> in the respiratory epithelium .


Leiomyosarcoma of the vulva : report of a case .


A 52-year-old female presented with a progressively enlarging vulvar mass .


Pathological evaluation revealed a high-grade vulvar leiomyosarcoma .


Immunohistochemical and ultrastructural studies were performed to support the diagnosis .


In an effort to better understand the biology of this tumor additional immunohistochemical studies for the protein product of p53 tumor suppressor gene and estrogen receptor expression by tumor cells , as well as the type of immune cells infiltrating the tumor were performed .

In an effort to better understand the biology of this tumor additional immunohistochemical studies for the <protein>protein product</protein> of <dna>p53 tumor suppressor gene</dna> and <protein>estrogen receptor</protein> expression by <cell_type>tumor cells</cell_type> , as well as the type of <cell_type>immune cells</cell_type> infiltrating the tumor were performed .


Tumor cells showed an overexpression of p53 protein and were estrogen receptor -positive .

<cell_type>Tumor cells</cell_type> showed an overexpression of <protein>p53 protein</protein> and were <protein>estrogen receptor</protein> -positive .


Macrophages and T and B lymphocytes infiltrated the tumor in moderate numbers with occasional lymphoid aggregate formation .

<cell_type>Macrophages</cell_type> and <cell_type>T and B lymphocytes</cell_type> infiltrated the tumor in moderate numbers with occasional lymphoid aggregate formation .


This study is the first attempt to better understand the biology of these tumors .


Stimulation of HIV replication in mononuclear phagocytes by leukemia inhibitory factor .

Stimulation of HIV replication in <cell_type>mononuclear phagocytes</cell_type> by <protein>leukemia inhibitory factor</protein> .


This study examined the effects of leukemia inhibitory factor ( LIF ) on human immunodeficiency virus ( HIV ) replication in mononuclear phagocytes ( MNP ) .

This study examined the effects of <protein>leukemia inhibitory factor</protein> ( <protein>LIF</protein> ) on human immunodeficiency virus ( HIV ) replication in <cell_type>mononuclear phagocytes</cell_type> ( <cell_type>MNP</cell_type> ) .


LIF induced a dose-dependent increase in p24 antigen production in the chronically infected promonocytic cell line U1 .

<protein>LIF</protein> induced a dose-dependent increase in <protein>p24 antigen</protein> production in the <cell_line>chronically infected promonocytic cell line U1</cell_line> .


The magnitude and time kinetics of the LIF effects were similar to interleukin 1 ( IL-1 ) , IL-6 , and tumor necrosis factor ( TNF ) , other cytokines known to induce HIV replication in this cell line .

The magnitude and time kinetics of the <protein>LIF</protein> effects were similar to <protein>interleukin 1</protein> ( <protein>IL-1</protein> ) , <protein>IL-6</protein> , and <protein>tumor necrosis factor</protein> ( <protein>TNF</protein> ) , other cytokines known to induce HIV replication in this cell line .


To characterize mechanisms responsible for these LIF effects , levels of HIV mRNA , activation of the DNA binding protein nuclear factor ( NF ) -kB , signal transduction pathways , and potential interactions with other cytokines were analyzed .

To characterize mechanisms responsible for these <protein>LIF</protein> effects , levels of <rna>HIV mRNA</rna> , activation of the <protein>DNA binding protein nuclear factor</protein> <protein>( NF ) -kB</protein> , signal transduction pathways , and potential interactions with other <protein>cytokines</protein> were analyzed .


LIF increased steady-state levels of HIV mRNA at 2.0 , 4.3 , and 9.2 kB .

<protein>LIF</protein> increased steady-state levels of <rna>HIV mRNA</rna> at 2.0 , 4.3 , and 9.2 kB .


This was detectable by 24 h and persisted until 72 h .


The DNA binding protein NF-kB is a central mediator in cytokine activation of HIV transcription .

The <protein>DNA binding protein NF-kB</protein> is a central mediator in <protein>cytokine</protein> activation of HIV transcription .


NF-kB levels were higher in unstimulated U1 cells as compared to the parent cell line U937 .

<protein>NF-kB</protein> levels were higher in <cell_line>unstimulated U1 cells</cell_line> as compared to the <cell_line>parent cell line</cell_line> <cell_line>U937</cell_line> .


In both cell lines LIF increased NF-kB activity .

In both cell lines <protein>LIF</protein> increased <protein>NF-kB</protein> activity .


Induction of NF-kB and HIV replication by cytokines are at least in part dependent on reactive oxygen intermediates .

Induction of <protein>NF-kB</protein> and HIV replication by <protein>cytokines</protein> are at least in part dependent on reactive oxygen intermediates .


The oxygen radical scavenger N-acetyl-L-cysteine , but not an inhibitor of nitric oxide synthase , inhibited LIF -induced HIV replication .

The oxygen radical scavenger N-acetyl-L-cysteine , but not an inhibitor of <protein>nitric oxide synthase</protein> , inhibited <protein>LIF</protein> -induced HIV replication .


LIF induces the production of other cytokines in monocytes but its effects on HIV replication were not inhibited by antibodies to IL-1 , TNF , or IL-6 .

<protein>LIF</protein> induces the production of other <protein>cytokines</protein> in monocytes but its effects on HIV replication were not inhibited by <protein>antibodies</protein> to <protein>IL-1</protein> , <protein>TNF</protein> , or <protein>IL-6</protein> .


These results identify LIF as a stimulus of HIV replication .

These results identify <protein>LIF</protein> as a stimulus of HIV replication .


( ABSTRACT TRUNCATED AT 250 WORDS )


Mechanisms involved in the inhibition of growth of a human B lymphoma cell line , B104 , by anti-MHC class II antibodies .

Mechanisms involved in the inhibition of growth of a <cell_line>human B lymphoma cell line</cell_line> , <cell_line>B104</cell_line> , by <protein>anti-MHC class II antibodies</protein> .


The mechanisms involved in the inhibition of growth of a human B lymphoma cell line , B104 , by anti-MHC class II antibodies ( Ab ) were compared with those in anti-IgM Ab -induced B104 growth inhibition .

The mechanisms involved in the inhibition of growth of a <cell_line>human B lymphoma cell line</cell_line> , <cell_line>B104</cell_line> , by <protein>anti-MHC class II antibodies</protein> ( <protein>Ab</protein> ) were compared with those in <protein>anti-IgM Ab</protein> -induced B104 growth inhibition .


Two anti-MHC class II Ab , L227 and 2.06 , inhibited the growth of B104 cells , although 2.06 , but not L227 , needed to be further cross-linked with a goat anti-mouse IgG Ab ( GAM ) to show the effect .

Two <protein>anti-MHC class II Ab</protein> , <protein>L227</protein> and <protein>2.06</protein> , inhibited the growth of <cell_line>B104 cells</cell_line> , although <protein>2.06</protein> , but not <protein>L227</protein> , needed to be further cross-linked with a <protein>goat anti-mouse IgG Ab</protein> ( <protein>GAM</protein> ) to show the effect .


L227 induced an increase in intracellular free Ca2+ concentration ( [ Ca2+ ] i ) from the intracellular pool and little or no protein tyrosine phosphorylation , phosphatidyl inositol turnover , or expression of Egr-1 mRNA , whereas 2.06 plus GAM induced an increase in [ Ca2+ ] i from both the intracellular and , in particular , the extracellular pools .

<protein>L227</protein> induced an increase in intracellular free Ca2+ concentration ( [ Ca2+ ] i ) from the intracellular pool and little or no protein tyrosine phosphorylation , phosphatidyl inositol turnover , or expression of <rna>Egr-1 mRNA</rna> , whereas <protein>2.06</protein> plus <protein>GAM</protein> induced an increase in [ Ca2+ ] i from both the intracellular and , in particular , the extracellular pools .


The inhibition of B104 cell growth induced by anti-MHC class II Ab was Ca ( 2+ ) -independent and not inhibited by actinomycin D or cyclosporin A , and cell cycle arrest at the G2/M interphase was not observed .

The inhibition of <cell_line>B104 cell</cell_line> growth induced by <protein>anti-MHC class II Ab</protein> was Ca ( 2+ ) -independent and not inhibited by actinomycin D or cyclosporin A , and cell cycle arrest at the G2/M interphase was not observed .


These features are very different from those observed in B104 cell death induced by anti-IgM Ab .

These features are very different from those observed in <cell_line>B104 cell</cell_line> death induced by <protein>anti-IgM Ab</protein> .


Neither DNA fragmentation nor the morphology of apoptosis was observed .


These findings demonstrate that cross-linking of MHC class II molecules transduced the negative signals through intracellular mechanisms different from those present in the cross-linking of surface IgM .

These findings demonstrate that cross-linking of <protein>MHC class II molecules</protein> transduced the negative signals through intracellular mechanisms different from those present in the cross-linking of surface <protein>IgM</protein> .


Functional block for 1 alpha , 25-dihydroxyvitamin D3-mediated gene regulation in human B lymphocytes .

Functional block for 1 alpha , 25-dihydroxyvitamin D3-mediated gene regulation in <cell_type>human B lymphocytes</cell_type> .


Elements necessary for the steroid hormone 1 alpha , 25-dihydroxyvitamin D3 ( 1 alpha , 25- ( OH ) 2D3 ) to induce a biological response include the presence of specific intracellular receptors ( vitamin D3 receptors ( VDR ) ) and modulation of gene expression via hormone-activated receptor binding to regulatory regions of target genes .

Elements necessary for the steroid hormone 1 alpha , 25-dihydroxyvitamin D3 ( 1 alpha , 25- ( OH ) 2D3 ) to induce a biological response include the presence of specific <protein>intracellular receptors</protein> ( <protein>vitamin D3 receptors</protein> ( <protein>VDR</protein> ) ) and modulation of gene expression via hormone-activated receptor binding to <protein>regulatory regions</protein> of <dna>target genes</dna> .


These parameters were examined in normal and Epstein-Barr virus-immortalized human B cells and compared with 1 alpha , 25- ( OH ) 2D3-responsive cells of the T and monocytic lineages .

These parameters were examined in normal and <cell_line>Epstein-Barr virus-immortalized human B cells</cell_line> and compared with <cell_line>1 alpha , 25- ( OH ) 2D3-responsive cells</cell_line> of the <cell_type>T and monocytic lineages</cell_type> .


Although resting tonsillar B cells did not express VDR mRNA , activation of these cells with interleukin-4 induced VDR in the absence of exogenously supplemented 1 alpha , 25- ( OH ) 2D3 .

Although resting tonsillar B cells did not express <rna>VDR mRNA</rna> , activation of these cells with <protein>interleukin-4</protein> induced <protein>VDR</protein> in the absence of exogenously supplemented 1 alpha , 25- ( OH ) 2D3 .


As indicators of hormone-mediated gene regulation we analyzed modulation of CD23 , a common B cell/monocyte surface antigen , and 24-hydroxylase .

As indicators of hormone-mediated gene regulation we analyzed modulation of <protein>CD23</protein> , a <protein>common B cell/monocyte surface antigen</protein> , and <protein>24-hydroxylase</protein> .


1 alpha , 25- ( OH ) 2D3 inhibited CD23 expression in U937 cells , yet failed to modulate CD23 expression in B cells .

1 alpha , 25- ( OH ) 2D3 inhibited <protein>CD23</protein> expression in <cell_line>U937 cells</cell_line> , yet failed to modulate <protein>CD23</protein> expression in <cell_type>B cells</cell_type> .


Furthermore , 1 alpha , 25- ( OH ) 2D3 induced 24-hydroxylase mRNA expression and metabolic activity in both U937 cells and lectin-activated T cells , yet failed to induce 24-hydroxylase mRNA or its metabolic activity in B cells .

Furthermore , 1 alpha , 25- ( OH ) 2D3 induced <protein>24-hydroxylase</protein> mRNA expression and metabolic activity in both <cell_line>U937 cells</cell_line> and <cell_type>lectin-activated T cells</cell_type> , yet failed to induce <rna>24-hydroxylase mRNA</rna> or its metabolic activity in <cell_type>B cells</cell_type> .


These findings suggest that although human B lymphocytes can express VDR mRNA and protein , they exhibit a functional block for vitamin D-dependent gene regulation .

These findings suggest that although <cell_type>human B lymphocytes</cell_type> can express <rna>VDR mRNA</rna> and <protein>protein</protein> , they exhibit a functional block for vitamin D-dependent gene regulation .


Positive and negative regulation of IL-2 gene expression : role of multiple regulatory sites .

Positive and negative regulation of <protein>IL-2</protein> gene expression : role of multiple regulatory sites .


Interleukin 2 ( IL-2 ) is an important lymphokine required in the process of T cell activation , proliferation , clonal expansion and differentiation .

Interleukin 2 ( <protein>IL-2</protein> ) is an important <protein>lymphokine</protein> required in the process of T cell activation , proliferation , clonal expansion and differentiation .


The IL-2 gene displays both T cell specific and inducible expression : it is only expressed in CD4+ T cells after antigenic or mitogenic stimulation .

The <dna>IL-2 gene</dna> displays both T cell specific and inducible expression : it is only expressed in <cell_type>CD4+ T cells</cell_type> after antigenic or mitogenic stimulation .


Several cis-acting regulatory sites are required for induction of the IL-2 gene after stimulation .

Several <dna>cis-acting regulatory sites</dna> are required for induction of the <dna>IL-2 gene</dna> after stimulation .


In this study , we have analysed the function of these cis-acting regulatory sites in the context of the native IL-2 enhancer and promoter sequence .

In this study , we have analysed the function of these <dna>cis-acting regulatory sites</dna> in the context of the native <dna>IL-2 enhancer</dna> and <dna>promoter sequence</dna> .


The results of this study suggest that the NFAT ( -276 to -261 ) , the distal octamer ( -256 to -248 ) and the proximal octamer ( -75 to -66 ) sites not only act as enhancers of IL-2 gene transcription in the presence of cellular stimulation , but also have a silencing effect on IL-2 gene expression in resting cells .

The results of this study suggest that the <protein>NFAT</protein> ( <dna>-276 to -261</dna> ) , the <dna>distal octamer</dna> ( <dna>-256 to -248</dna> ) and the <dna>proximal octamer</dna> ( <dna>-75 to -66</dna> ) sites not only act as enhancers of <dna>IL-2 gene</dna> transcription in the presence of cellular stimulation , but also have a silencing effect on <dna>IL-2 gene</dna> expression in resting cells .


Two other sites display disparate effects on IL-2 gene expression in different T leukemia cell lines : the distal purine box ( -291 to -277 ) and the proximal purine box sites ( -145 to -128 ) .

Two other sites display disparate effects on <dna>IL-2 gene</dna> expression in different <cell_line>T leukemia cell lines</cell_line> : the <dna>distal purine box</dna> ( <dna>-291 to -277</dna> ) and the <dna>proximal purine box sites</dna> ( <dna>-145 to -128</dna> ) .


Finally , the AP-1 ( -186 to -176 ) and the kappa B sites ( -206 to -195 ) respond to different cellular activation in EL4 cells .

Finally , the <dna>AP-1 ( -186 to -176 ) and the kappa B sites</dna> ( <dna>-206 to -195</dna> ) respond to different cellular activation in <cell_line>EL4 cells</cell_line> .


The AP-1 site mediated the response to PMA stimulation while the kappa B site responded to IL-1 stimulation .

The <dna>AP-1 site</dna> mediated the response to PMA stimulation while the <dna>kappa B site</dna> responded to <protein>IL-1</protein> stimulation .


These data suggest that the regulation of IL-2 gene expression is a complex process and multiple cis-acting regulatory sites interact to exert different effects in T cells representative of alternative stages of differentiation .

These data suggest that the regulation of <dna>IL-2 gene</dna> expression is a complex process and multiple <dna>cis-acting regulatory sites</dna> interact to exert different effects in <cell_type>T cells</cell_type> representative of alternative stages of differentiation .


Sp1 is a critical factor for the monocytic specific expression of human CD14 .

<protein>Sp1</protein> is a critical factor for the monocytic specific expression of <protein>human CD14</protein> .


CD14 is a membrane glycoprotein expressed specifically on monocytes and macrophages , and its expression is markedly increased during the process of monocyte differentiation .

<protein>CD14</protein> is a <protein>membrane glycoprotein</protein> expressed specifically on <cell_type>monocytes</cell_type> and <cell_type>macrophages</cell_type> , and its expression is markedly increased during the process of <cell_type>monocyte</cell_type> differentiation .


In order to study CD14 gene regulation , the human CD14 gene was cloned from a partial EcoRI digested chromosome 5 library .

In order to study <dna>CD14 gene</dna> regulation , the <dna>human CD14 gene</dna> was cloned from a partial <dna>EcoRI digested chromosome 5 library</dna> .


A 5.5-kilobase genomic clone contained the full-length CD14 coding sequence and 4.2 kilobases of 5'-upstream sequence .

A <dna>5.5-kilobase genomic clone</dna> contained the full-length <dna>CD14 coding sequence</dna> and <dna>4.2 kilobases of 5'-upstream sequence</dna> .


One major and one minor transcription start site were identified 101 and 130 base pairs ( bp ) upstream , respectively , from the protein translation start ATG .

One <dna>major and one minor transcription start site</dna> were identified <dna>101 and 130 base pairs ( bp ) upstream</dna> , respectively , from the <dna>protein translation start ATG</dna> .


A DNA fragment containing 128 bp of upstream sequence had strong , monocyte-specific promoter activity in the CD14 positive monocytic cell line Mono Mac 6 as compared to the nonmonocytic cell lines HeLa and REX .

A <dna>DNA fragment</dna> containing <dna>128 bp of upstream sequence</dna> had strong , monocyte-specific promoter activity in the <cell_line>CD14 positive monocytic cell line</cell_line> <cell_line>Mono Mac 6</cell_line> as compared to the <cell_line>nonmonocytic cell lines</cell_line> <cell_line>HeLa</cell_line> and <cell_line>REX</cell_line> .


Four regions in this DNA fragment interact with nuclear proteins isolated from monocytic cells .

Four regions in this <dna>DNA fragment</dna> interact with <protein>nuclear proteins</protein> isolated from <cell_type>monocytic cells</cell_type> .


The Sp1 transcription factor bound to three different regions in the CD14 promoter .

The <protein>Sp1</protein> <protein>transcription factor</protein> bound to three different regions in the <dna>CD14 promoter</dna> .


Mutation of the major Sp1 binding site ( -110 bp ) decreased tissue-specific promoter activity , and these results , together with transactivation experiments , demonstrate that Sp1 plays a critical role in the tissue-specific expression of CD14 in monocytic cells .

Mutation of the <dna>major Sp1 binding site</dna> ( <dna>-110 bp</dna> ) decreased tissue-specific promoter activity , and these results , together with transactivation experiments , demonstrate that <protein>Sp1</protein> plays a critical role in the tissue-specific expression of <protein>CD14</protein> in <cell_type>monocytic cells</cell_type> .


CD14 Sp1 site oligonucleotides bound preferentially to a 105-kDa Sp1 species , which is present in higher relative levels in monocytic than non-monocytic cells , suggesting that modification of Sp1 , such as phosphorylation , may explain how the Sp1 site mediates monocytic specific promoter activity .

<protein>CD14</protein> <protein>Sp1</protein> site oligonucleotides bound preferentially to a <protein>105-kDa Sp1 species</protein> , which is present in higher relative levels in <cell_type>monocytic</cell_type> than <cell_type>non-monocytic cells</cell_type> , suggesting that modification of <protein>Sp1</protein> , such as phosphorylation , may explain how the <protein>Sp1</protein> site mediates monocytic specific promoter activity .


The interleukin-8 AP-1 and kappa B-like sites are genetic end targets of FK506-sensitive pathway accompanied by calcium mobilization .

The <dna>interleukin-8 AP-1 and kappa B-like sites</dna> are genetic end targets of FK506-sensitive pathway accompanied by calcium mobilization .


FK506 , an immunosuppressant , inhibits the production of several cytokines in T lymphocytes .

FK506 , an immunosuppressant , inhibits the production of several <protein>cytokines</protein> in <cell_type>T lymphocytes</cell_type> .


We observed that FK506 suppressed the transcription of a chemotactic cytokine , interleukin-8 ( IL-8 ) in a human T cell line , Jurkat cells , activated by phorbol 12-myristate 13-acetate ( PMA ) and calcium ( Ca2+ ) ionophore ( ionomycin ) .

We observed that FK506 suppressed the transcription of a <protein>chemotactic cytokine</protein> , <protein>interleukin-8</protein> ( <protein>IL-8</protein> ) in a <cell_line>human T cell line</cell_line> , <cell_line>Jurkat cells</cell_line> , activated by phorbol 12-myristate 13-acetate ( PMA ) and calcium ( Ca2+ ) ionophore ( ionomycin ) .


By deleted and mutated analysis of the IL-8 promoters , the AP-1 and kappa B-like sites were identified as the responsive elements for PMA and ionomycin .

By deleted and mutated analysis of the <dna>IL-8 promoters</dna> , the <dna>AP-1 and kappa B-like sites</dna> were identified as the responsive elements for PMA and ionomycin .


FK506 suppressed the transcriptions through the AP-1 or kappa B-like sites induced by PMA plus Ca ( 2+ ) -mobilizing agents , but not those induced by Ca ( 2+ ) -independent stimuli .

FK506 suppressed the transcriptions through the <dna>AP-1</dna> or <dna>kappa B-like sites</dna> induced by PMA plus Ca ( 2+ ) -mobilizing agents , but not those induced by Ca ( 2+ ) -independent stimuli .


In gel retardation analysis , FK506 had little effect on the binding to the AP-1 site of PMA/ionomycin-induced nuclear factors , which were recognized with anti-JunD or c-Fos antibody .

In gel retardation analysis , FK506 had little effect on the binding to the <dna>AP-1 site</dna> of PMA/ionomycin-induced nuclear factors , which were recognized with <protein>anti-JunD</protein> or <protein>c-Fos antibody</protein> .


In contrast , FK506 or EGTA ( Ca2+ chelator ) similarly affected the formation of kappa B-like site binding complexes , which were not recognized by any antibodies against the human Rel family proteins ( c-Rel , p65 , p50 , and p49 ) .

In contrast , FK506 or EGTA ( Ca2+ chelator ) similarly affected the formation of <protein>kappa B-like site binding complexes</protein> , which were not recognized by any <protein>antibodies</protein> against the <protein>human Rel family proteins</protein> ( <protein>c-Rel</protein> , <protein>p65</protein> , <protein>p50</protein> , and <protein>p49</protein> ) .


Furthermore , we confirmed the previous report that FK506 suppressed the PMA/ionomycin-induced activation through authentic kappa B site of immunoglobulin ( Ig ) gene , to which NF-kappa B binding was also decreased by FK506 , indicating that both IL-8 kappa B-like site and Ig kappa B site are FK506-sensitive in spite of the difference of binding factors .

Furthermore , we confirmed the previous report that FK506 suppressed the PMA/ionomycin-induced activation through authentic <dna>kappa B site</dna> of <dna>immunoglobulin ( Ig ) gene</dna> , to which <protein>NF-kappa B</protein> binding was also decreased by FK506 , indicating that both <dna>IL-8 kappa B-like site</dna> and Ig <dna>kappa B site</dna> are FK506-sensitive in spite of the difference of <protein>binding factors</protein> .


Our results indicate that not only the reported IL-2 NF-AT and NFIL-2A sites and Ig kappa B site , but also the IL-8 AP-1 and kappa B-like sites are terminals of FK506-sensitive pathway involving Ca2+ mobilization .

Our results indicate that not only the reported <dna>IL-2 NF-AT and NFIL-2A sites</dna> and Ig <dna>kappa B site</dna> , but also the <protein>IL-8</protein> <dna>AP-1</dna> and <dna>kappa B-like sites</dna> are terminals of FK506-sensitive pathway involving Ca2+ mobilization .


Androgen binding sites in peripheral human mononuclear leukocytes of healthy males and females .

<protein>Androgen binding sites</protein> in <cell_type>peripheral human mononuclear leukocytes</cell_type> of healthy males and females .


Androgen binding sites have been identified in circulating human mononuclear leukocytes of healthy donors of both sexes .

<protein>Androgen binding sites</protein> have been identified in <cell_type>circulating human mononuclear leukocytes</cell_type> of healthy donors of both sexes .


Cells were separated from blood samples on a Ficoll gradient and incubated with different concentrations of [ 3H ] testosterone in the presence or absence of a 400-fold excess of unlabelled testosterone .


Binding data were derived from Scatchard analysis .


The binding sites fulfil the required criteria for specific steroid binding sites however differ somewhat from the classic androgen receptors from genital skin fibroblast : in fertile adult males ( n = 20 ) the binding sites showed ( 1 ) a high affinity for testosterone ( 1.32 +/- 0.49 nM ; mean +/- SD ) , ( 2 ) a saturable capacity ( 184 +/- 52 binding sites per cell ; mean +/- SD ) , and ( 3 ) a characteristic competitive binding profile for other steroid hormones ( relative binding affinities : testosterone = dihydrotestosterone > 17 beta-estradiol > progesterone , whereas aldosterone , 17-hydroxy-progesterone and cortisol did not compete appreciably ) .

The <protein>binding sites</protein> fulfil the required criteria for specific <protein>steroid binding sites</protein> however differ somewhat from the classic <protein>androgen receptors</protein> from <cell_type>genital skin fibroblast</cell_type> : in fertile adult males ( n = 20 ) the <protein>binding sites</protein> showed ( 1 ) a high affinity for testosterone ( 1.32 +/- 0.49 nM ; mean +/- SD ) , ( 2 ) a saturable capacity ( 184 +/- 52 <protein>binding sites</protein> per cell ; mean +/- SD ) , and ( 3 ) a characteristic competitive binding profile for other steroid hormones ( relative binding affinities : testosterone = dihydrotestosterone > 17 beta-estradiol > progesterone , whereas aldosterone , 17-hydroxy-progesterone and cortisol did not compete appreciably ) .


Furthermore the number of binding sites determined using [ 3H ] dihydrotestosterone , [ 3H ] RU-1881 , or [ 3H ] testosterone were comparable .

Furthermore the number of <protein>binding sites</protein> determined using [ 3H ] dihydrotestosterone , [ 3H ] RU-1881 , or [ 3H ] testosterone were comparable .


This raises the possibility that androgen receptors in peripheral mononuclear leukocytes differ from those in genital skin fibroblasts .

This raises the possibility that <protein>androgen receptors</protein> in <cell_type>peripheral mononuclear leukocytes</cell_type> differ from those in <cell_type>genital skin fibroblasts</cell_type> .


There was no apparent correlation between serum testosterone concentrations and androgen binding sites .

There was no apparent correlation between serum testosterone concentrations and <protein>androgen binding sites</protein> .


In fertile women remarkable changes in androgen binding sites were seen in the course of the menstrual cycle , with a significant increase in the immediate preovulatory period .

In fertile women remarkable changes in <protein>androgen binding sites</protein> were seen in the course of the menstrual cycle , with a significant increase in the immediate preovulatory period .


The presence of androgen receptors in peripheral mononuclear leukocytes provides for the first time the experimental basis for an hypothesis of direct , receptor-mediated effects of androgens on mature immunocompetent cells .

The presence of <protein>androgen receptors</protein> in <cell_type>peripheral mononuclear leukocytes</cell_type> provides for the first time the experimental basis for an hypothesis of direct , receptor-mediated effects of androgens on <cell_type>mature immunocompetent cells</cell_type> .


The immunological implications of these results are discussed .


Induction of IL-8 expression in T cells uses the CD28 costimulatory pathway .

Induction of <protein>IL-8</protein> expression in <cell_type>T cells</cell_type> uses the <protein>CD28</protein> costimulatory pathway .


IL-8 , a potent chemotactic factor for neutrophil granulocytes and lymphocytes , is a proinflammatory cytokine secreted by a variety of cell types , including T cells .

<protein>IL-8</protein> , a potent <protein>chemotactic factor</protein> for <cell_type>neutrophil granulocytes</cell_type> and <cell_type>lymphocytes</cell_type> , is a proinflammatory cytokine secreted by a variety of cell types , including <cell_type>T cells</cell_type> .


Stimulation of the CD28 cell surface molecule delivers costimulatory signals essential for lymphokine production in activated T cells via a conserved sequence element found in the promoter of several lymphokine genes .

Stimulation of the <protein>CD28 cell surface molecule</protein> delivers costimulatory signals essential for <protein>lymphokine</protein> production in <cell_type>activated T cells</cell_type> via a <dna>conserved sequence element</dna> found in the <dna>promoter</dna> of several <dna>lymphokine genes</dna> .


Anti-CD28-stimulated T cells produced significant amounts of IL-8 ; additionally , costimulation with anti-CD3 and anti-CD28 Abs resulted in a synergistic induction of IL-8 secretion .

<cell_line>Anti-CD28-stimulated T cells</cell_line> produced significant amounts of <protein>IL-8</protein> ; additionally , costimulation with <protein>anti-CD3</protein> and <protein>anti-CD28 Abs</protein> resulted in a synergistic induction of <protein>IL-8</protein> secretion .


Sequence homology , single nucleotide mutations , and anti-CD28 Ab stimulation studies established that the NF-kappa B-like sequence in the promoter of the IL-8 gene functioned as a CD28 response element .

Sequence homology , single nucleotide mutations , and <protein>anti-CD28 Ab</protein> stimulation studies established that the <dna>NF-kappa B-like sequence</dna> in the <dna>promoter</dna> of the <dna>IL-8 gene</dna> functioned as a <dna>CD28 response element</dna> .


Furthermore , cyclosporin A , but not rapamycin , blocked the synergistic induction of IL-8 expression achieved with anti-CD3 and anti- CD28 costimulation .

Furthermore , cyclosporin A , but not rapamycin , blocked the synergistic induction of <protein>IL-8</protein> expression achieved with <protein>anti-CD3</protein> and anti- <protein>CD28</protein> costimulation .


The involvement of a CD28 response element in the induction of IL-8 expression in activated T cells may provide new insights into the pathogenesis and persistence of immune disorders characterized by increased levels of IL-8 , such as psoriasis and rheumatoid arthritis .

The involvement of a <dna>CD28 response element</dna> in the induction of <protein>IL-8</protein> expression in <cell_type>activated T cells</cell_type> may provide new insights into the pathogenesis and persistence of immune disorders characterized by increased levels of <protein>IL-8</protein> , such as psoriasis and rheumatoid arthritis .


MHC class II signaling in B-cell activation [ see comments ]

<protein>MHC class II</protein> signaling in <cell_type>B-cell</cell_type> activation [ see comments ]


The cognate interaction between T cells and antigen-presenting cells ( APCs ) , mediated by major histocompatibility complex ( MHC ) class II molecules , results in the delivery of activation signals to the APC .

The cognate interaction between <cell_type>T cells</cell_type> and <cell_type>antigen-presenting cells</cell_type> ( <cell_type>APCs</cell_type> ) , mediated by <protein>major histocompatibility complex ( MHC ) class II molecules</protein> , results in the delivery of activation signals to the <cell_type>APC</cell_type> .


These signals contribute to the expression of co-stimulatory activity by APCs and have important consequences for cell effector function .

These signals contribute to the expression of co-stimulatory activity by <cell_type>APCs</cell_type> and have important consequences for cell effector function .


MHC class II molecules also serve as receptors for B-cell stimulation by microbial superantigens .

<protein>MHC class II</protein> molecules also serve as receptors for <cell_type>B-cell</cell_type> stimulation by <protein>microbial superantigens</protein> .


In this review , Paul Scholl and Raif Geha discuss recent advances in our understanding of mechanisms of MHC class II signaling and analyse their role in human B-cell activation .

In this review , Paul Scholl and Raif Geha discuss recent advances in our understanding of mechanisms of <protein>MHC class II</protein> signaling and analyse their role in human <cell_type>B-cell</cell_type> activation .


Effects of glucocorticoids in rheumatoid arthritis .


Diminished glucocorticoid receptors do not result in glucocorticoid resistance .

Diminished <protein>glucocorticoid receptors</protein> do not result in glucocorticoid resistance .


OBJECTIVE .


Lymphocytes of patients with rheumatoid arthritis ( RA ) have diminished receptor density ; thus , patients with RA should show partial resistance to glucocorticoids .


We investigated the glucocorticoid sensitivity of lymphocytes in RA patients compared with healthy subjects .

We investigated the glucocorticoid sensitivity of <cell_type>lymphocytes</cell_type> in RA patients compared with healthy subjects .


METHODS .


We determined the effects of glucocorticoids on lymphocyte proliferation and cytokine release .

We determined the effects of glucocorticoids on <cell_type>lymphocyte</cell_type> proliferation and <protein>cytokine</protein> release .


RESULTS .


Proliferation and cytokine release were inhibited in RA patients to the same extent as in healthy controls .

Proliferation and <protein>cytokine</protein> release were inhibited in RA patients to the same extent as in healthy controls .


CONCLUSION .


Diminished receptor density in RA patients does not result in glucocorticoid resistance .


Arrested development : understanding v-abl .

Arrested development : understanding <dna>v-abl</dna> .


The protein tyrosine kinase activity of the v-abl oncogene has been demonstrated to subvert the normal second messenger systems used by lymphoid cells for growth and differentiation .

The <protein>protein tyrosine kinase</protein> activity of the <dna>v-abl oncogene</dna> has been demonstrated to subvert the normal second messenger systems used by <cell_type>lymphoid cells</cell_type> for growth and differentiation .


Transformation of bone marrow with the Abelson murine leukemia virus results in the appearance of B cell lineage cells arrested at the pre-B cell stage .

Transformation of bone marrow with the Abelson murine leukemia virus results in the appearance of <cell_type>B cell lineage cells</cell_type> arrested at the pre-B cell stage .


Recent reports have characterized these cells expressing high v-abl kinase activity as deficient in detectable NF-kappaB DNA binding activity and low level RAG gene expression .

Recent reports have characterized these cells expressing high <dna>v-abl</dna> kinase activity as deficient in detectable <protein>NF-kappaB</protein> DNA binding activity and low level <dna>RAG gene</dna> expression .


These observations suggest that v-abl may be inhibiting the differentiation of B cells by blocking these two crucial elements in the maturation pathway .

These observations suggest that <dna>v-abl</dna> may be inhibiting the differentiation of <cell_type>B cells</cell_type> by blocking these two crucial elements in the maturation pathway .


Corticosteroid receptors in lymphocytes : a possible marker of brain involution ?

<protein>Corticosteroid receptors</protein> in <cell_type>lymphocytes</cell_type> : a possible marker of brain involution ?


A similarity has recently been found between the regulation of corticosteroid receptors in brain and in lymphoid tissue .

A similarity has recently been found between the regulation of <protein>corticosteroid receptors</protein> in brain and in lymphoid tissue .


We have studied the regulation of corticosteroid receptors in human mononuclear leukocytes as a possible marker of brain involution .

We have studied the regulation of <protein>corticosteroid receptors</protein> in <cell_type>human mononuclear leukocytes</cell_type> as a possible marker of brain involution .


Type I corticosteroid receptors are down regulated by excess of mineralocorticoids ( primary and secondary hyperaldosteronism , pseudohyperaldosteronism ) and of glucocorticoids ( Cushing 's syndrome ) . Type II corticosteroid receptors are not reduced by excess of endogenous corticosteroids ( Cushing 's syndrome ) .

<protein>Type I corticosteroid receptors</protein> are down regulated by excess of mineralocorticoids ( primary and secondary hyperaldosteronism , pseudohyperaldosteronism ) and of glucocorticoids ( Cushing 's syndrome ) . <protein>Type II corticosteroid receptors</protein> are not reduced by excess of endogenous corticosteroids ( Cushing 's syndrome ) .


In normal adults there is a direct significant correlation between plasma cortisol and Type I and between plasma cortisol and Type II receptors in mononuclear leukocytes , while in Cushing 's syndrome the correlation is inverse between plasma cortisol at 8 a.m. and Type II receptors .

In normal adults there is a direct significant correlation between plasma cortisol and <protein>Type I</protein> and between plasma cortisol and <protein>Type II receptors</protein> in <cell_type>mononuclear leukocytes</cell_type> , while in Cushing 's syndrome the correlation is inverse between plasma cortisol at 8 a.m. and <protein>Type II receptors</protein> .


In an aged population the mean numbers of Type I and of Type II receptors are lower and plasma cortisol is higher than in adult controls , but the increase of plasma cortisol is not followed by a clinical picture of hypercorticism .

In an aged population the mean numbers of <protein>Type I</protein> and of <protein>Type II receptors</protein> are lower and plasma cortisol is higher than in adult controls , but the increase of plasma cortisol is not followed by a clinical picture of hypercorticism .


Corticosteroid Type I and Type II receptors are inversely correlated with age .

<protein>Corticosteroid Type I and Type II receptors</protein> are inversely correlated with age .


After dexamethasone suppression ( 1 mg at 11 p.m. ) Type I receptors always decrease in controls while the response of Type II is not homogeneous .

After dexamethasone suppression ( 1 mg at 11 p.m. ) <protein>Type I receptors</protein> always decrease in controls while the response of <protein>Type II</protein> is not homogeneous .


In an aged group of patients , both receptors are reduced by dexamethasone .


We conclude that the decrease with age of corticosteroid receptors is possibly related to a physiological involution of corticosteroid receptors and that this reduction does increase plasma cortisol concentration , without affecting the glucocorticoid effector mechanism .

We conclude that the decrease with age of <protein>corticosteroid receptors</protein> is possibly related to a physiological involution of <protein>corticosteroid receptors</protein> and that this reduction does increase plasma cortisol concentration , without affecting the glucocorticoid effector mechanism .


All-trans retinoic acid and 1 alpha , 25-dihydroxyvitamin D3 co-operate to promote differentiation of the human promyeloid leukemia cell line HL60 to monocytes .

All-trans retinoic acid and 1 alpha , 25-dihydroxyvitamin D3 co-operate to promote differentiation of the <cell_line>human promyeloid leukemia cell line</cell_line> <cell_line>HL60</cell_line> to <cell_type>monocytes</cell_type> .


A basis for differentiation therapy of leukemias is provided by knowledge of agents which induce specific lineage maturation .


All-trans retinoic acid ( RA ) induces differentiation of HL60 cells to neutrophils and is used to treat acute promyelocytic leukemia .

All-trans retinoic acid ( RA ) induces differentiation of <cell_line>HL60 cells</cell_line> to <cell_type>neutrophils</cell_type> and is used to treat acute promyelocytic leukemia .


We observed that RA did not induced neutrophil differentiation in serum-free grown HL60 cells whereas 50 nM 1 alpha , 25-dihydroxyvitamin D3 ( D3 ) induced maximal monocyte differentiation .

We observed that RA did not induced neutrophil differentiation in <cell_line>serum-free grown HL60 cells</cell_line> whereas 50 nM 1 alpha , 25-dihydroxyvitamin D3 ( D3 ) induced maximal monocyte differentiation .


Increasing RA concentrations reduced the D3 concentration required for monocyte differentiation .


Cells treated with 5 nM D3 showed little response , but differentiated maximally with 5 nM D3 and 10 nM RA .


The D3 analogs MC903 , EB1089 and KH1060 were more potent inducers of monocyte differentiation .


The extent to which analog activity was increased after cotreatment with RA was inversely related to potency .


Twenty-four hour treatment with 10 nM RA primed cells for response to 5 nM D3 ; the reverse sequence being ineffective .

Twenty-four hour treatment with 10 nM <cell_line>RA primed cells</cell_line> for response to 5 nM D3 ; the reverse sequence being ineffective .


Priming with 10 nM RA , or subsequent treatment with D3 ( 5 nM ) , did not alter expression of mRNAs encoding receptors for D3 ( VDR ) , RA ( RAR alpha ) or 9-CIS RA ( RXR alpha , beta , gamma ) .

Priming with 10 nM RA , or subsequent treatment with D3 ( 5 nM ) , did not alter expression of <rna>mRNAs</rna> encoding <protein>receptors</protein> for D3 ( <protein>VDR</protein> ) , RA ( <protein>RAR alpha</protein> ) or 9-CIS RA ( <protein>RXR alpha , beta , gamma</protein> ) .


That RA promotes both neutrophil and monocyte differentiation has implications for the use of RA and D3 in treatment of leukemias and provides insight into mechanisms whereby RAR , VDR and RXR facilitate monocyte differentiation .

That RA promotes both neutrophil and monocyte differentiation has implications for the use of RA and D3 in treatment of leukemias and provides insight into mechanisms whereby RAR , <protein>VDR</protein> and <protein>RXR</protein> facilitate monocyte differentiation .


[ An overexpression of retinoic acid receptor alpha blocks myeloid cell differentiation at the promyelocyte stage ]

[ An overexpression of <protein>retinoic acid receptor alpha</protein> blocks <cell_type>myeloid cell</cell_type> differentiation at the promyelocyte stage ]


Retinoic acid ( RA ) , a vitamin A derivative , exerts a wide range of biological effects related to cell proliferation and differentiation .


The pleiotropic effects of RA are thought to be mediated through specific nuclear RA receptors ( RARs ) .

The pleiotropic effects of RA are thought to be mediated through specific nuclear <protein>RA receptors</protein> ( <protein>RARs</protein> ) .


RARs are members of the steroid/thyroid hormone receptor superfamily and exhibit a molecular structure that possess discrete DNA-binding and RA ( ligand ) -binding domains .

<protein>RARs</protein> are members of the <protein>steroid/thyroid hormone receptor superfamily</protein> and exhibit a molecular structure that possess discrete DNA-binding and <protein>RA ( ligand ) -binding domains</protein> .


In hematopoietic system , RA and RARs , predominantly RAR alpha may play key roles for the proliferation and differentiation of hematopoietic progenitors .

In hematopoietic system , RA and <protein>RARs</protein> , predominantly <protein>RAR alpha</protein> may play key roles for the proliferation and differentiation of <cell_type>hematopoietic progenitors</cell_type> .


However , it is currently unknown how RA and RARs are involved in regulating normal hematopoietic differentiation .

However , it is currently unknown how RA and <protein>RARs</protein> are involved in regulating normal hematopoietic differentiation .


To make clear the roles of RA and RAR alpha in the normal hematopoiesis , I have introduced the construct of human RAR alpha ( hRAR alpha ) into murine bone marrow cells with retroviral vector , and selected infected cells with drug resistant marker ( Neo ( r ) ) cultured on the stroma cell line ( PA6-neo ) , and analyzed the behavior of infected cells .

To make clear the roles of RA and <protein>RAR alpha</protein> in the normal hematopoiesis , I have introduced the construct of <protein>human RAR alpha</protein> ( <protein>hRAR alpha</protein> ) into <cell_type>murine bone marrow cells</cell_type> with retroviral vector , and selected infected cells with <dna>drug resistant marker</dna> ( <dna>Neo ( r )</dna> ) cultured on the <cell_line>stroma cell line</cell_line> ( <cell_line>PA6-neo</cell_line> ) , and analyzed the behavior of <cell_type>infected cells</cell_type> .


All of procedure were done in vitro .


Most cells infected with hRAR alpha exhibited promyelocytic morphology and were thought to be blocked at the promyelocytic stage in their myeloid differentiation .

Most cells infected with <protein>hRAR alpha</protein> exhibited <cell_type>promyelocytic morphology</cell_type> and were thought to be blocked at the promyelocytic stage in their myeloid differentiation .


Furthermore , these immature cells differentiated terminally into mature granulocytes by adding with RA ( 10 ( -6 ) M ) .

Furthermore , these <cell_type>immature cells</cell_type> differentiated terminally into mature granulocytes by adding with RA ( 10 ( -6 ) M ) .


RAR alpha infected cells were also able to differentiate into mature macrophages in the both of long term culture and IL3 colony .

<protein>RAR alpha</protein> <cell_type>infected cells</cell_type> were also able to differentiate into <cell_type>mature macrophages</cell_type> in the both of long term culture and <cell_line>IL3 colony</cell_line> .


These observations suggest that an overexpression of RAR alpha alone is effective to suppress myeloid cell differentiation and RAR alpha plays a crucial role in the terminal differentiation of myeloid precursors .

These observations suggest that an overexpression of <protein>RAR alpha</protein> alone is effective to suppress <cell_type>myeloid cell</cell_type> differentiation and <protein>RAR alpha</protein> plays a crucial role in the terminal differentiation of myeloid precursors .


The system described here may serve as a model for studying the the essential genes for differentiation of normal bone marrow cells .

The system described here may serve as a model for studying the the <dna>essential genes</dna> for differentiation of <cell_type>normal bone marrow cells</cell_type> .


Hypoxic induction of interleukin-8 gene expression in human endothelial cells .

Hypoxic induction of <dna>interleukin-8 gene</dna> expression in <cell_type>human endothelial cells</cell_type> .


Because leukocyte-mediated tissue damage is an important component of the pathologic picture in ischemia/reperfusion , we have sought mechanisms by which PMNs are directed into hypoxic tissue .

Because leukocyte-mediated tissue damage is an important component of the pathologic picture in ischemia/reperfusion , we have sought mechanisms by which <cell_type>PMNs</cell_type> are directed into hypoxic tissue .


Incubation of human endothelial cells ( ECs ) in hypoxia , PO2 approximately 14-18 Torr , led to time-dependent release of IL-8 antigen into the conditioned medium ; this was accompanied by increased chemotactic activity for PMNs , blocked by antibody to IL-8 .

Incubation of <cell_type>human endothelial cells</cell_type> ( <cell_type>ECs</cell_type> ) in hypoxia , PO2 approximately 14-18 Torr , led to time-dependent release of <protein>IL-8</protein> antigen into the conditioned medium ; this was accompanied by increased chemotactic activity for <cell_type>PMNs</cell_type> , blocked by antibody to <protein>IL-8</protein> .


Production of IL-8 by hypoxic ECs occurred concomitantly with both increased levels of IL-8 mRNA , based on polymerase chain reaction analysis , and increased IL-8 transcription , based on nuclear run-on assays .

Production of <protein>IL-8</protein> by hypoxic <cell_type>ECs</cell_type> occurred concomitantly with both increased levels of <rna>IL-8 mRNA</rna> , based on polymerase chain reaction analysis , and increased <protein>IL-8</protein> transcription , based on nuclear run-on assays .


Northern analysis of mRNA from hypoxic ECs also demonstrated increased levels of mRNA for macrophage chemotactic protein-1 , another member of the chemokine superfamily of proinflammatory cytokines .

Northern analysis of mRNA from hypoxic <cell_type>ECs</cell_type> also demonstrated increased levels of <rna>mRNA</rna> for <protein>macrophage chemotactic protein-1</protein> , another member of the <protein>chemokine superfamily</protein> of <protein>proinflammatory cytokines</protein> .


IL-8 gene induction was associated with the presence of increased binding activity in nuclear extracts from hypoxic ECs for the NF-kB site .

<dna>IL-8 gene</dna> induction was associated with the presence of increased binding activity in nuclear extracts from <cell_type>hypoxic ECs</cell_type> for the NF-kB site .


Studies with human umbilical vein segments exposed to hypoxia also demonstrated increased elaboration of IL-8 antigen compared with normoxic controls .

Studies with human umbilical vein segments exposed to hypoxia also demonstrated increased elaboration of <protein>IL-8 antigen</protein> compared with normoxic controls .


In mice exposed to hypoxia ( PO2 approximately 30-40 Torr ) , there was increased pulmonary leukostasis , as evidenced by increased myeloperoxidase activity in tissue homogenates .

In mice exposed to hypoxia ( PO2 approximately 30-40 Torr ) , there was increased pulmonary leukostasis , as evidenced by increased <protein>myeloperoxidase</protein> activity in tissue homogenates .


In parallel , increased levels of transcripts for IP-10 , a murine homologue in the chemokine family related to IL-8 , were observed in hypoxic lung tissue .

In parallel , increased levels of transcripts for <protein>IP-10</protein> , a murine homologue in the <protein>chemokine family</protein> related to <protein>IL-8</protein> , were observed in hypoxic lung tissue .


Taken together , these data suggest that hypoxia constitutes a stimulus for leukocyte chemotaxis and tissue leukostasis .


Thrombin and thrombin receptor agonist peptide induce early events of T cell activation and synergize with TCR cross-linking for CD69 expression and interleukin 2 production .

<protein>Thrombin</protein> and <protein>thrombin receptor</protein> agonist peptide induce early events of T cell activation and synergize with <protein>TCR</protein> cross-linking for <protein>CD69</protein> expression and interleukin 2 production .


Thrombin stimulation of the T leukemic cell line Jurkat induced a transient increase in [ Ca2+ ] i .

<protein>Thrombin</protein> stimulation of the <cell_line>T leukemic cell line Jurkat</cell_line> induced a transient increase in [ Ca2+ ] i .


Proteolytic activity of the enzyme was required for this effect since diisopropyl fluorophosphate-thrombin failed to increase [ Ca2+ ] i .

Proteolytic activity of the <protein>enzyme</protein> was required for this effect since <protein>diisopropyl fluorophosphate-thrombin</protein> failed to increase [ Ca2+ ] i .


Furthermore , hirudin and anti-thrombin III inhibited the thrombin-induced [ Ca2+ ] i rise in Jurkat T cells .

Furthermore , hirudin and <protein>anti-thrombin III</protein> inhibited the thrombin-induced [ Ca2+ ] i rise in <cell_line>Jurkat T cells</cell_line> .


A synthetic thrombin receptor agonist peptide ( TRP ) of 7 residues ( SFLLRNP ) was found to be as effective as thrombin for [ Ca2+ ] i mobilization , and both agonists induced Ca2+ release exclusively from internal stores .

A synthetic <protein>thrombin receptor</protein> agonist peptide ( TRP ) of 7 residues ( SFLLRNP ) was found to be as effective as <protein>thrombin</protein> for [ Ca2+ ] i mobilization , and both agonists induced Ca2+ release exclusively from internal stores .


Thrombin stimulated tyrosine phosphorylation of several proteins of molecular mass 40 , 42 , 70 , 120 , and 130 kDa .

<protein>Thrombin</protein> stimulated tyrosine phosphorylation of several proteins of <protein>molecular mass 40 , 42 , 70 , 120 , and 130 kDa</protein> .


There was a good correlation between thrombin -induced tyrosine phosphorylation of the latter three proteins and Ca2+ mobilization .

There was a good correlation between <protein>thrombin</protein> -induced tyrosine phosphorylation of the latter three proteins and Ca2+ mobilization .


Thrombin and TRP also caused translocation of protein kinase C from the cytosol to the plasma membrane .

<protein>Thrombin</protein> and TRP also caused translocation of <protein>protein kinase C</protein> from the cytosol to the plasma membrane .


As a likely consequence of these events , thrombin activated the nuclear factor NF-kB .

As a likely consequence of these events , <protein>thrombin</protein> activated the <protein>nuclear factor NF-kB</protein> .


Several cell lines of hematopoietic origin including the leukemic T cell line HPB.ALL and the erythroleukemic cell line K562 were responsive to thrombin , whereas others such as THP1 , a myelomonocytic cell line , and BL2 , a Burkitt lymphoma were refractory to thrombin or TRP stimulation .

Several cell lines of hematopoietic origin including the <cell_line>leukemic T cell line HPB.ALL</cell_line> and the <cell_line>erythroleukemic cell line K562</cell_line> were responsive to <protein>thrombin</protein> , whereas others such as <cell_line>THP1</cell_line> , a <cell_line>myelomonocytic cell line</cell_line> , and <cell_line>BL2 ,</cell_line> a <cell_line>Burkitt lymphoma</cell_line> were refractory to <protein>thrombin</protein> or TRP stimulation .


The magnitude of the thrombin response in the different cell types paralleled the expression of the thrombin receptor mRNA .

The magnitude of the <protein>thrombin</protein> response in the different cell types paralleled the expression of the <rna>thrombin receptor mRNA</rna> .


We found that activation of Jurkat T cells by a combination of phytohemagglutinin and phorbol 12-myristate 13-acetate led to a dramatic inhibition of thrombin receptor mRNA expression and to a concomitant loss of the thrombin response .

We found that activation of <cell_line>Jurkat T cells</cell_line> by a combination of <protein>phytohemagglutinin</protein> and phorbol 12-myristate 13-acetate led to a dramatic inhibition of <rna>thrombin receptor mRNA</rna> expression and to a concomitant loss of the <protein>thrombin</protein> response .


Finally , we demonstrate that thrombin and TRP enhanced CD69 expression and interleukin 2 production induced by T cell receptor cross-linking in both Jurkat T cells and peripheral blood lymphocytes .

Finally , we demonstrate that <protein>thrombin</protein> and TRP enhanced <protein>CD69</protein> expression and <protein>interleukin 2</protein> production induced by <protein>T cell receptor</protein> cross-linking in both <cell_line>Jurkat T cells</cell_line> and peripheral blood <cell_type>lymphocytes</cell_type> .


These findings highlight the role of thrombin as a potential regulator of T lymphocyte activation .

These findings highlight the role of <protein>thrombin</protein> as a potential regulator of <cell_type>T lymphocyte</cell_type> activation .


Stress response of senescent T lymphocytes : reduced hsp70 is independent of the proliferative block .

Stress response of <cell_type>senescent T lymphocytes</cell_type> : reduced <protein>hsp70</protein> is independent of the proliferative block .


Senescent human T lymphocyte cultures are unable to undergo proliferation , but show no difference from early passage cells in cytotoxic function or surface antigenic profile .

<cell_line>Senescent human T lymphocyte cultures</cell_line> are unable to undergo proliferation , but show no difference from <cell_line>early passage cells</cell_line> in cytotoxic function or surface antigenic profile .


A second feature of senescent T cells is the dramatic reduction in hsp70 production in response to heat shock .

A second feature of <cell_line>senescent T cells</cell_line> is the dramatic reduction in <protein>hsp70</protein> production in response to heat shock .


This decline is associated with a decrease in binding of nuclear extracts to the consensus heat shock element .

This decline is associated with a decrease in binding of nuclear extracts to the <dna>consensus heat shock element</dna> .


Interestingly , the progressive decline in the heat shock response of cultured T cells correlates with the percent proliferative life span completed rather than with the actual proliferative activity at the time of heat shock .

Interestingly , the progressive decline in the heat shock response of <cell_line>cultured T cells</cell_line> correlates with the percent proliferative life span completed rather than with the actual proliferative activity at the time of heat shock .


This suggests that for senescent T cells the reduced ability to respond to heat shock by producing hsp70 , although possibly lying at the level of transcriptional control , may nevertheless be unrelated to the reduced DNA synthesis or the diminished proliferative activity also manifested by these cells .

This suggests that for <cell_type>senescent T cells</cell_type> the reduced ability to respond to heat shock by producing <protein>hsp70</protein> , although possibly lying at the level of transcriptional control , may nevertheless be unrelated to the reduced DNA synthesis or the diminished proliferative activity also manifested by these cells .


Involvement of phospholipase D in the activation of transcription factor AP-1 in human T lymphoid Jurkat cells .

Involvement of <protein>phospholipase D</protein> in the activation of <protein>transcription factor AP-1</protein> in <cell_line>human T lymphoid Jurkat cells</cell_line> .


The induction of the AP-1 transcription factor has been ascribed to the early events leading to T lymphocyte activation .

The induction of the <protein>AP-1</protein> <protein>transcription factor</protein> has been ascribed to the early events leading to T lymphocyte activation .


We have examined the possibility that stimulation of phospholipase D ( PLD ) may regulate activation of transcription factor AP-1 in human T cells by transfecting human T lymphocyte Jurkat cells with a plasmid containing an AP-1 enhancer element and a chloramphenicol acetyltransferase reporter gene .

We have examined the possibility that stimulation of <protein>phospholipase D</protein> ( <protein>PLD</protein> ) may regulate activation of <protein>transcription factor AP-1</protein> in human T cells by transfecting human <cell_line>T lymphocyte Jurkat cells</cell_line> with a plasmid containing an <dna>AP-1 enhancer element</dna> and a <dna>chloramphenicol acetyltransferase reporter gene</dna> .


We have detected activatable PLD in Jurkat cells , and we have found that addition of phosphatidic acid ( PA ) , the physiologic product of PLD action on phospholipids , is rapidly incorporated into Jurkat cells and leads to activation of transcription factor AP-1 .

We have detected activatable <protein>PLD</protein> in <cell_line>Jurkat cells</cell_line> , and we have found that addition of phosphatidic acid ( PA ) , the physiologic product of <protein>PLD</protein> action on phospholipids , is rapidly incorporated into <cell_line>Jurkat cells</cell_line> and leads to activation of <protein>transcription factor AP-1</protein> .


Treatment of Jurkat cells with anti-CD3 mAb activated both PLD and transcription factor AP-1 .

Treatment of <cell_line>Jurkat cells</cell_line> with <protein>anti-CD3 mAb</protein> activated both <protein>PLD</protein> and <protein>transcription factor AP-1</protein> .


Wortmannin , an inhibitor of receptor-coupled PLD activation , blocked the anti-CD3 -induced increases in both PLD activity and AP-1 enhancer activity .

Wortmannin , an inhibitor of receptor-coupled <protein>PLD</protein> activation , blocked the <protein>anti-CD3</protein> -induced increases in both <protein>PLD</protein> activity and <dna>AP-1 enhancer</dna> activity .


We found a good correlation in the transfected cells between PLD activation and induction of AP-1 enhancer activity under different experimental conditions .

We found a good correlation in the <cell_line>transfected cells</cell_line> between <protein>PLD</protein> activation and induction of <dna>AP-1 enhancer</dna> activity under different experimental conditions .


Furthermore , ethanol , an inhibitor of the PLD pathway , blocked the anti-CD3-stimulated AP-1 enhancer activity .

Furthermore , ethanol , an inhibitor of the <protein>PLD</protein> pathway , blocked the anti-CD3-stimulated <dna>AP-1 enhancer</dna> activity .


However , this anti-CD3 -mediated response was not inhibited by neomycin , an inhibitor of phosphoinositide hydrolysis .

However , this <protein>anti-CD3</protein> -mediated response was not inhibited by neomycin , an inhibitor of phosphoinositide hydrolysis .


The increases in AP-1 enhancer activity induced by PA or anti-CD3 mAb were efficiently abrogated by the presence of propranolol , an inhibitor of PA phosphohydrolase and protein kinase C ( PKC ) .

The increases in <dna>AP-1 enhancer</dna> activity induced by PA or <protein>anti-CD3 mAb</protein> were efficiently abrogated by the presence of propranolol , an inhibitor of PA <protein>phosphohydrolase</protein> and <protein>protein kinase C</protein> ( <protein>PKC</protein> ) .


Furthermore , the PA -and the anti-CD3 -induced increases in AP-1 enhancer activity were blocked by the presence of PKC inhibitors or by PKC down-regulation .

Furthermore , the PA -and the <protein>anti-CD3</protein> -induced increases in <dna>AP-1 enhancer</dna> activity were blocked by the presence of <protein>PKC</protein> inhibitors or by <protein>PKC</protein> down-regulation .