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Hla variants of human lympho blastoid cell lines as targets for cyto toxic t cells analysis of cyto toxic t lymphocytes reactivity against 2 hla dr variants

Hla variants of human lympho blastoid cell lines as targets for cyto toxic t cells analysis of cyto toxic t lymphocytes reactivity against 2 hla dr variants

Journal of Immunology 125(6): 2515-2520

To test whether HLA-D-associated determinants can restrict cytotoxic activity, hapten-specific CTL [cytotoxic T lymphocytes] were raised by co-culture of peripheral blood lymphocytes raised by co-culture of peripheral with TNP[trinitrophenyl]-modified autologous cells. Effector cells were tested against autologous cells, the B lymphoblastoid cell line T5-1 and 2 HLA-DR variants of T5-1 derived by immunoselection. T5-1 has a normal complement of HLA-A, B, C antigens and expresses the DR1/ DR3 specificities. A partial HLA-D loss variant, 816, and a total HLA-D loss variant, 616, were tested along with the parent. T5-1, 616 and 816 were used as 51Cr-labeled targets or as unlabeled target competitors of the lysis of autologous cells. The 816 variant, deficient in Dr1 expression, was comparable to the parent as a target in cross-reactive lysis. The 616 variant, lacking membrane expression of all HLA-D-associated antigens, was on average only 70% as susceptible as the parent line. Effector cells from 31 individuals representing HLA-DR types 1-7, 10 and 11 were tested. Some individuals showed a much better ability than others to distinguish the parent and 616 variant. In ability to inhibit lysis of autologous T cell blasts, the toal HLA-D loss variant (616) was on average only 60% as effective as the parent cell line. Some individuals were better able than others to distinguish T5-1 from 616 but this ability did not appear to segregate with a particular HLA type. Controls showed that the variant was not resistant to T cell-mediated lysis and that the 3 B lymphoblastoid cell lines possessed equivalent levels of HLA A, B and C antigens. They all coupled with .apprx. 55 .times. 107 molecules of TNP/cell. Full expression of the 29,000/34,000-dalton HLA-D heterodimer required for maximal lysis and such specificity may explain some of the widespread hapten-specific CTL cross-reactivity.

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Accession: 005583304

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