We previously observed that pseudorabies virus (PRV)-induced, cell-mediated cytolysis in pigs includes killing by natural killer (NK) cells. We also observed that IL-2 stimulation of naive PBMC expands porcine NK cells. The purpose of this study was to compare the phenotypes of the cytolytic subsets stimulated by PRV and by IL-2. PBMC were isolated from blood of PRV-immune and naive pigs and stimulated with PRV or IL-2. After 6 days, the frequency of various lymphocyte subsets in these cultured PBMC was determined by flow cytometry: the cells were separated with a magnet-activated cell sorter and the cytolytic activity of the separated populations was determined. When lymphocytes were separated and analysed with FACScan, the following lymphocyte subsets were discriminated: CD6 CD8 CD4 (CTL phenotype), CD6 CD8 CD4 (the fraction containing memory T helper cells), CD6 CD8 CD4 (T helper cell phenotype), CD6 CD8 CD4 γδ-T ( γδ-T cell phenotype), CD6 CD8 CD4 γδ-T (NK phenotype) and CD6 CD8 CD4 γδ-T or  γδ-T. Flow cytometry analysis demonstrated that PRV stimulation of immune PBMC resulted in the occurrence of more CD6 CD8 and CD4 CD8 and fewer CD6 CD8 and γδ-T CD8 lymphocytes than IL-2 stimulation of naive PBMC (<0·05). It was demonstrated further that killing by PRV-stimulated PBMC was mediated mainly by CD6 CD8 T lymphocytes. Killing by IL-2-stimulated PBMC was mediated mainly by CD6 CD8 T lymphocytes. These results demonstrate that both natural killing and killing by classical PRV-specific CTL were detected in PRV-immune pigs, whereas IL-2 stimulation of PBMC isolated from naive pigs mainly induced natural killing.


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