HIV-Nef enhances interleukin-2 production and phosphatidylinositol 3-kinase activity in a human T cell line
The Nef protein has a major influence on disease pathogenesis in HIV-infected individuals. The objective of the present study was to examine the effects of Nef on T lymphocyte activation and associated signalling events. A recombinant vaccinia expression system was used to express Nef in a human T c...
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Veröffentlicht in: | AIDS (London) 2000-08, Vol.14 (12), p.1701-1707 |
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description | The Nef protein has a major influence on disease pathogenesis in HIV-infected individuals. The objective of the present study was to examine the effects of Nef on T lymphocyte activation and associated signalling events.
A recombinant vaccinia expression system was used to express Nef in a human T cell line. Stimulation of these cells with anti-CD28 antibody, and either phorbol 12-myristate 13-acetate (PMA) or anti-CD3, activates signal transduction pathways and results in IL-2 production and IL-2 receptor alpha-chain (CD25) expression. Cellular responses were examined in cells expressing either Nef or an irrelevant control protein.
Activation of signalling was assessed by immunoblot analysis, or by in-vitro phosphatidylinositol 3-kinase (PI3K) assays. IL-2 production was measured by enzyme-linked immunosorbent assay, and CD25 cell surface expression was examined using flow cytometry.
Infection of cells with recombinant vaccinia expressing HIV-nef resulted in a marked increase in the production of IL-2 when cells were activated. The enhanced IL-2 response was accompanied by an increase in the level of PI3K activity. IL-2 production remained sensitive to inhibition with the PI3K competitive inhibitor Ly294002, and to the fungal macrolide, rapamycin. In contrast, CD25 expression was not affected, and there were no measurable changes to nuclear factor kappaB (NFkappaB) activation pathways.
Enhanced IL-2 production in stimulated T cells expressing HIV-Nef is associated with increased activation of PI3K-dependent signalling pathways. The results support a model in which Nef affects HIV disease progression by distorting T cell responses. |
doi_str_mv | 10.1097/00002030-200008180-00003 |
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A recombinant vaccinia expression system was used to express Nef in a human T cell line. Stimulation of these cells with anti-CD28 antibody, and either phorbol 12-myristate 13-acetate (PMA) or anti-CD3, activates signal transduction pathways and results in IL-2 production and IL-2 receptor alpha-chain (CD25) expression. Cellular responses were examined in cells expressing either Nef or an irrelevant control protein.
Activation of signalling was assessed by immunoblot analysis, or by in-vitro phosphatidylinositol 3-kinase (PI3K) assays. IL-2 production was measured by enzyme-linked immunosorbent assay, and CD25 cell surface expression was examined using flow cytometry.
Infection of cells with recombinant vaccinia expressing HIV-nef resulted in a marked increase in the production of IL-2 when cells were activated. The enhanced IL-2 response was accompanied by an increase in the level of PI3K activity. IL-2 production remained sensitive to inhibition with the PI3K competitive inhibitor Ly294002, and to the fungal macrolide, rapamycin. In contrast, CD25 expression was not affected, and there were no measurable changes to nuclear factor kappaB (NFkappaB) activation pathways.
Enhanced IL-2 production in stimulated T cells expressing HIV-Nef is associated with increased activation of PI3K-dependent signalling pathways. The results support a model in which Nef affects HIV disease progression by distorting T cell responses.</description><identifier>ISSN: 0269-9370</identifier><identifier>EISSN: 1473-5571</identifier><identifier>DOI: 10.1097/00002030-200008180-00003</identifier><identifier>PMID: 10985305</identifier><language>eng</language><publisher>Hagerstown, MD: Lippincott Williams & Wilkins</publisher><subject>AIDS/HIV ; Biological and medical sciences ; CD25 antigen ; CD28 Antigens ; Enzyme Activation ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Gene Expression Regulation, Viral ; Gene Products, nef - genetics ; Gene Products, nef - physiology ; Genes, nef - physiology ; HIV-1 - genetics ; Human immunodeficiency virus ; Human viral diseases ; Humans ; Immunoblotting ; Infectious diseases ; Interleukin-2 - biosynthesis ; Interleukin-2 - metabolism ; Jurkat Cells ; Medical sciences ; nef Gene Products, Human Immunodeficiency Virus ; Nef protein ; Phosphatidylinositol 3-Kinases - metabolism ; Receptors, Interleukin-2 - biosynthesis ; Receptors, Interleukin-2 - immunology ; Signal Transduction ; T-Lymphocytes - enzymology ; T-Lymphocytes - immunology ; Viral diseases ; Viral diseases of the lymphoid tissue and the blood. Aids</subject><ispartof>AIDS (London), 2000-08, Vol.14 (12), p.1701-1707</ispartof><rights>2000 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c421t-ff6471ade8ddad3ced35464c01b3a012683b3c714ae3fa62b9fbca36934372233</citedby><cites>FETCH-LOGICAL-c421t-ff6471ade8ddad3ced35464c01b3a012683b3c714ae3fa62b9fbca36934372233</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1512993$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10985305$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>SCHIBECI, S. D</creatorcontrib><creatorcontrib>CLEGG, A. O</creatorcontrib><creatorcontrib>BITI, R. A</creatorcontrib><creatorcontrib>SAGAWA, K</creatorcontrib><creatorcontrib>STEWART, G. J</creatorcontrib><creatorcontrib>WILLIAMSON, P</creatorcontrib><title>HIV-Nef enhances interleukin-2 production and phosphatidylinositol 3-kinase activity in a human T cell line</title><title>AIDS (London)</title><addtitle>AIDS</addtitle><description>The Nef protein has a major influence on disease pathogenesis in HIV-infected individuals. The objective of the present study was to examine the effects of Nef on T lymphocyte activation and associated signalling events.
A recombinant vaccinia expression system was used to express Nef in a human T cell line. Stimulation of these cells with anti-CD28 antibody, and either phorbol 12-myristate 13-acetate (PMA) or anti-CD3, activates signal transduction pathways and results in IL-2 production and IL-2 receptor alpha-chain (CD25) expression. Cellular responses were examined in cells expressing either Nef or an irrelevant control protein.
Activation of signalling was assessed by immunoblot analysis, or by in-vitro phosphatidylinositol 3-kinase (PI3K) assays. IL-2 production was measured by enzyme-linked immunosorbent assay, and CD25 cell surface expression was examined using flow cytometry.
Infection of cells with recombinant vaccinia expressing HIV-nef resulted in a marked increase in the production of IL-2 when cells were activated. The enhanced IL-2 response was accompanied by an increase in the level of PI3K activity. IL-2 production remained sensitive to inhibition with the PI3K competitive inhibitor Ly294002, and to the fungal macrolide, rapamycin. In contrast, CD25 expression was not affected, and there were no measurable changes to nuclear factor kappaB (NFkappaB) activation pathways.
Enhanced IL-2 production in stimulated T cells expressing HIV-Nef is associated with increased activation of PI3K-dependent signalling pathways. The results support a model in which Nef affects HIV disease progression by distorting T cell responses.</description><subject>AIDS/HIV</subject><subject>Biological and medical sciences</subject><subject>CD25 antigen</subject><subject>CD28 Antigens</subject><subject>Enzyme Activation</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Flow Cytometry</subject><subject>Gene Expression Regulation, Viral</subject><subject>Gene Products, nef - genetics</subject><subject>Gene Products, nef - physiology</subject><subject>Genes, nef - physiology</subject><subject>HIV-1 - genetics</subject><subject>Human immunodeficiency virus</subject><subject>Human viral diseases</subject><subject>Humans</subject><subject>Immunoblotting</subject><subject>Infectious diseases</subject><subject>Interleukin-2 - biosynthesis</subject><subject>Interleukin-2 - metabolism</subject><subject>Jurkat Cells</subject><subject>Medical sciences</subject><subject>nef Gene Products, Human Immunodeficiency Virus</subject><subject>Nef protein</subject><subject>Phosphatidylinositol 3-Kinases - metabolism</subject><subject>Receptors, Interleukin-2 - biosynthesis</subject><subject>Receptors, Interleukin-2 - immunology</subject><subject>Signal Transduction</subject><subject>T-Lymphocytes - enzymology</subject><subject>T-Lymphocytes - immunology</subject><subject>Viral diseases</subject><subject>Viral diseases of the lymphoid tissue and the blood. Aids</subject><issn>0269-9370</issn><issn>1473-5571</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkctKxDAUhoMoznh5BclC3EWTnF7SpQzqCINu1G05TVIm2knHphXm7U2d8bIzBHIIX_788BFCBb8UvMiveFySA2dynJRQnI0D7JGpSHJgaZqLfTLlMitYATmfkKMQXiORcqUOySSGqBR4OiVv8_sX9mBrav0SvbaBOt_brrHDm_NM0nXXmkH3rvUUvaHrZRvWS-yd2TTOt8H1bUOBRRaDpRjBD9dvYgZFuhxW6OkT1bZpaKTtCTmosQn2dHcek-fbm6fZnC0e7-5n1wumEyl6VtdZkgs0VhmDBrQ1kCZZormoALmQmYIKdC4StFBjJquirjRCVkACuZQAx-RimxvLvw829OXKhbEFetsOoYxQyiHu_0CRZypRRRZBtQV114bQ2bpcd26F3aYUvByNlN9Gyh8jX1djmbPdH0O1subPw62CCJzvAAwam7qLGlz45VIhiwLgE8iXk0g</recordid><startdate>20000818</startdate><enddate>20000818</enddate><creator>SCHIBECI, S. D</creator><creator>CLEGG, A. O</creator><creator>BITI, R. A</creator><creator>SAGAWA, K</creator><creator>STEWART, G. J</creator><creator>WILLIAMSON, P</creator><general>Lippincott Williams & Wilkins</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20000818</creationdate><title>HIV-Nef enhances interleukin-2 production and phosphatidylinositol 3-kinase activity in a human T cell line</title><author>SCHIBECI, S. D ; CLEGG, A. O ; BITI, R. A ; SAGAWA, K ; STEWART, G. J ; WILLIAMSON, P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c421t-ff6471ade8ddad3ced35464c01b3a012683b3c714ae3fa62b9fbca36934372233</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>AIDS/HIV</topic><topic>Biological and medical sciences</topic><topic>CD25 antigen</topic><topic>CD28 Antigens</topic><topic>Enzyme Activation</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Flow Cytometry</topic><topic>Gene Expression Regulation, Viral</topic><topic>Gene Products, nef - genetics</topic><topic>Gene Products, nef - physiology</topic><topic>Genes, nef - physiology</topic><topic>HIV-1 - genetics</topic><topic>Human immunodeficiency virus</topic><topic>Human viral diseases</topic><topic>Humans</topic><topic>Immunoblotting</topic><topic>Infectious diseases</topic><topic>Interleukin-2 - biosynthesis</topic><topic>Interleukin-2 - metabolism</topic><topic>Jurkat Cells</topic><topic>Medical sciences</topic><topic>nef Gene Products, Human Immunodeficiency Virus</topic><topic>Nef protein</topic><topic>Phosphatidylinositol 3-Kinases - metabolism</topic><topic>Receptors, Interleukin-2 - biosynthesis</topic><topic>Receptors, Interleukin-2 - immunology</topic><topic>Signal Transduction</topic><topic>T-Lymphocytes - enzymology</topic><topic>T-Lymphocytes - immunology</topic><topic>Viral diseases</topic><topic>Viral diseases of the lymphoid tissue and the blood. Aids</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>SCHIBECI, S. D</creatorcontrib><creatorcontrib>CLEGG, A. O</creatorcontrib><creatorcontrib>BITI, R. A</creatorcontrib><creatorcontrib>SAGAWA, K</creatorcontrib><creatorcontrib>STEWART, G. J</creatorcontrib><creatorcontrib>WILLIAMSON, P</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>AIDS (London)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>SCHIBECI, S. D</au><au>CLEGG, A. O</au><au>BITI, R. A</au><au>SAGAWA, K</au><au>STEWART, G. J</au><au>WILLIAMSON, P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>HIV-Nef enhances interleukin-2 production and phosphatidylinositol 3-kinase activity in a human T cell line</atitle><jtitle>AIDS (London)</jtitle><addtitle>AIDS</addtitle><date>2000-08-18</date><risdate>2000</risdate><volume>14</volume><issue>12</issue><spage>1701</spage><epage>1707</epage><pages>1701-1707</pages><issn>0269-9370</issn><eissn>1473-5571</eissn><abstract>The Nef protein has a major influence on disease pathogenesis in HIV-infected individuals. The objective of the present study was to examine the effects of Nef on T lymphocyte activation and associated signalling events.
A recombinant vaccinia expression system was used to express Nef in a human T cell line. Stimulation of these cells with anti-CD28 antibody, and either phorbol 12-myristate 13-acetate (PMA) or anti-CD3, activates signal transduction pathways and results in IL-2 production and IL-2 receptor alpha-chain (CD25) expression. Cellular responses were examined in cells expressing either Nef or an irrelevant control protein.
Activation of signalling was assessed by immunoblot analysis, or by in-vitro phosphatidylinositol 3-kinase (PI3K) assays. IL-2 production was measured by enzyme-linked immunosorbent assay, and CD25 cell surface expression was examined using flow cytometry.
Infection of cells with recombinant vaccinia expressing HIV-nef resulted in a marked increase in the production of IL-2 when cells were activated. The enhanced IL-2 response was accompanied by an increase in the level of PI3K activity. IL-2 production remained sensitive to inhibition with the PI3K competitive inhibitor Ly294002, and to the fungal macrolide, rapamycin. In contrast, CD25 expression was not affected, and there were no measurable changes to nuclear factor kappaB (NFkappaB) activation pathways.
Enhanced IL-2 production in stimulated T cells expressing HIV-Nef is associated with increased activation of PI3K-dependent signalling pathways. The results support a model in which Nef affects HIV disease progression by distorting T cell responses.</abstract><cop>Hagerstown, MD</cop><pub>Lippincott Williams & Wilkins</pub><pmid>10985305</pmid><doi>10.1097/00002030-200008180-00003</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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subjects | AIDS/HIV Biological and medical sciences CD25 antigen CD28 Antigens Enzyme Activation Enzyme-Linked Immunosorbent Assay Flow Cytometry Gene Expression Regulation, Viral Gene Products, nef - genetics Gene Products, nef - physiology Genes, nef - physiology HIV-1 - genetics Human immunodeficiency virus Human viral diseases Humans Immunoblotting Infectious diseases Interleukin-2 - biosynthesis Interleukin-2 - metabolism Jurkat Cells Medical sciences nef Gene Products, Human Immunodeficiency Virus Nef protein Phosphatidylinositol 3-Kinases - metabolism Receptors, Interleukin-2 - biosynthesis Receptors, Interleukin-2 - immunology Signal Transduction T-Lymphocytes - enzymology T-Lymphocytes - immunology Viral diseases Viral diseases of the lymphoid tissue and the blood. Aids |
title | HIV-Nef enhances interleukin-2 production and phosphatidylinositol 3-kinase activity in a human T cell line |
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