title: Costimulation of human CD4+ T cells with LFA-3 and B7 induce distinct effects on AP-1 and NF-kappa B transcription factors.
abstract: We have earlier shown that stimulation of human CD4+ T cells with SEA presented on Chinese hamster ovary (CHO)-DR transfectants coexpressing either B7 or LFA-3 resulted in distinct cytokine profiles. We now demonstrate that B7, but not LFA-3, strongly costimulated IL-2 transcription and mRNA expression in CD4+ T cells. Maximal increase in IL-2 transcription was recorded with CHO-DR/B7/LFA-3, suggesting a cooperative effect of B7 and LFA-3 at the transcriptional level. Gel-shift analysis demonstrated that stimulation of CD4+ T cells with CHO-DR and staphylococcal enterotoxin A was sufficient to induce significant amounts of NF-kappa B binding proteins, whereas induction of AP-1 binding proteins required costimulation. LFA-3 induced moderate levels of AP-1, but did not influence the levels of NF-kappa B, while B7 costimulation strongly induced both AP-1 and substantially enhanced NF-kappa B binding proteins. The CHO-DR/B7/LFA-3 triple transfectant induced a further increase in AP-1 and NF-kappa B binding proteins compared with the double transfectants. The level of Oct-1 binding proteins remained similar in all samples. Super-shift analysis revealed that the NF-kappa B complex of costimulated CD4+ T cells contained large amounts of p50, substantial amounts of p65, and marginal levels of c-Rel proteins. The AP-1 binding proteins contained c-Jun, Jun-D, and Fra-1, but marginal amounts of Jun-B and c-Fos. Our results indicate distinct effects of B7 and LFA-3 costimulation on the activity of AP-1 and NF-kappa B. These may partly account for the differential effects of B7 and LFA-3 costimulation on IL-2 expression.
authors.affiliation: Wallenberg Laboratory, Department of Tumor Immunology, University of Lund, Sweden.
authors.forename: E
authors.initials: E
authors.lastname: Parra
authors.forename: M
authors.initials: M
authors.lastname: Varga
authors.forename: M
authors.initials: M
authors.lastname: Sigvardsson
authors.forename: G
authors.initials: G
authors.lastname: Hedlund
authors.forename: T
authors.initials: T
authors.lastname: Kalland
authors.forename: T
authors.initials: T
authors.lastname: Leanderson
authors.forename: H O
authors.initials: HO
authors.lastname: Sjögren
authors.forename: M
authors.initials: M
authors.lastname: Dohlsten
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000502:physiology*; D018122:B7-1 Antigen / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018968:CD58 Antigens; D016466:CHO Cells; D002455:Cell Division; D016384:Consensus Sequence; D006224:Cricetinae; D004268:DNA-Binding Proteins / Q000378:metabolism; D004768:Enterotoxins / Q000276:immunology; D005786:Gene Expression Regulation / Q000502:physiology*; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology; D051863:Host Cell Factor C1; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D050811:Octamer Transcription Factor-1; D011994:Recombinant Proteins / Q000276:immunology; D015398:Signal Transduction / Q000502:physiology; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D015703:Antigens, CD; D018122:B7-1 Antigen; D018968:CD58 Antigens; D004268:DNA-Binding Proteins; D004768:Enterotoxins; C497692:HCFC1 protein, human; D006684:HLA-DR Antigens; D051863:Host Cell Factor C1; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D050811:Octamer Transcription Factor-1; C494165:POU2F1 protein, human; D011994:Recombinant Proteins; D018808:Transcription Factor AP-1; D014157:Transcription Factors; C049463:enterotoxin E, Staphylococcal; C026066:enterotoxin A, Staphylococcal