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The Journal of Immunology, Vol 155, Issue 3 1132-1140, Copyright © 1995 by American Association of Immunologists

ARTICLES

Costimulation of human CD4+ T cells with LFA-3 and B7 induce distinct effects on AP-1 and NF-kappa B transcription factors

E Parra, M Varga, M Sigvardsson, G Hedlund, T Kalland, T Leanderson, HO Sjogren and M Dohlsten
Wallenberg Laboratory, Department of Tumor Immunology, University of Lund, Sweden.

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.


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E. Parra, K. McGuire, G. Hedlund, and M. Dohlsten
Overexpression of p65 and c-Jun Substitutes for B7-1 Costimulation by Targeting the CD28RE Within the IL-2 Promoter
J. Immunol., June 1, 1998; 160(11): 5374 - 5381.
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