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The Journal of Immunology, Vol 156, Issue 5 1848-1855, Copyright © 1996 by American Association of Immunologists

ARTICLES

Mutations in human CD4 impair the functional interaction with different human and mouse class II isotypes and alleles

S Fleury, B Huang, A Zerbib, G Croteau, EO Long and RP Sekaly
Immunology Laboratory, Montreal Clinical Research Institute, Canada.

The structure-function of the CD4-class II MHC interaction was investigated. Two functional assays were used to assess the responses of the 3DT52.5.8 murine T cell hybridoma expressing human CD4 (h-CD4) or murine CD4 (m-CD4). First, we determined the responses of the CD4+ and CD4-effector cells toward DAP-3 cells co-expressing the cognate alloantigen H-2Dd together with several human (DRw52b, DR4-Dw4, DR2A, and DPw2) and murine (I-Ab, I-Ak, IA alpha b I-A beta k and I-Ek) class II alleles and isotypes. We found that h-CD4 and m-CD4 strongly enhance the T cell response to H-2Dd, demonstrating that interspecies CD4/class II interactions occur efficiently. Furthermore, mutations in h-CD4 at positions 19, 89, and 165 markedly reduced the interaction with both human class II and mouse class II, indicating that the structural features of this cross-species interaction are strongly conserved. This was further supported by the finding that a h-CD4 deletion mutant (deletion F43-S49) interacted with both human and murine class II. Moreover, as 3DT cells express the responsive V beta element for the bacterial superantigen staphylococcal enterotoxin B, a co-receptor assay was conducted. DAP-3 cells expressing only class II molecules were used as APCs to present staphylococcal enterotoxin B to h-CD4+ and m-CD4+ T cells. h-CD4 and m-CD4 were able to enhance the T cell response to staphylococcal enterotoxin B, further demonstrating the conservation of the CD4-class II MHC interaction.


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