To determine the specificity, if any, of cellular cytotoxicity against transmissible gastro-enteritis virus (TGEV) infected cells, we developed a test using B lymphoblasts from a MHC histocompatible (d/d haplotype) cell line (L14), as stimulating and target cells. These cells were previously infected with recombinant vaccinia virus including different TGEV structural genes, either the spike (vS), membrane (vM) or nucleoprotein gene (vN). Lymphocytes from a TGEV immunized (d/d) swine developed a cytotoxic activity after secondary in vitro stimulation in the presence of vS, vM or vN infected L14 cells. The cytotoxic activity was induced and directed against the homologous vS and vM infected cells but no cytotoxic activity occurred at all against vN infected cells. While vM infected cells induced a cytotoxic activity against vM infected cells only, vS infected cells stimulated a cross-reactive cytotoxic activity against vM and vN infected cells in addition to that against vS infected cells. This latter cytotoxicity may be due to an increase in a non-specific background of Natural Killer or lymphocyte activated killer activity, which is seen also after coculture with wild type vaccinia virus (vW) infected cells. Thus these results are of practical importance in two respects. First, lymphoïd B cell lines represent an excellent tool for determining which viral antigens are recognized by cytotoxic lymphocytes and second, they indicate the need to incorporate the M and S genes into a TGEV vaccine to induce cellular immunity against TGEV.

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