Objective To analyze the functional differences between virus-specific CD4+T cells and CD8+T cells in patients infected with Epstein-Barr virus(EBV)who develop liver injury and those who do not.Methods 45 cases of EBV infections were enrolled,including 28 cases developing liver injuries and 17 that did not.Mononuclear cells from peripheral blood were isolated.CD4+T cells and CD8+T cells were purified and cultured using recombinant EBV core antigen 2(EBNA2)for 96 h with stimulation.The CCK-8 method was used to detect cell proliferation.Flow cytometry was used to detect the proportion of CD4+T cells and CD8+T cells.An enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of CD4+T cells secreting cytokines and CD8+T cells secreting molecular toxicity.Real-time quantitative PCR was used to detect the mRNA levels of transcription factors and molecular toxicity in CD4+T cell subsets.Flow cytometry was used to detect the immune checkpoints at molecular levels in CD8+T cells.The inter-group comparison was performed using a t-test or Mann-Whitney test.Results There was no statistically significant difference(P＞0.05)in the proliferation proportion of peripheral blood mononuclear cells,CD4+T cells,and CD8+T cells after stimulation with recombinant EBNA2 between the EBV-infected non-liver injury group and the infected liver injury group(P＞0.05).There was no statistically significant difference in the proportion of CD4+T cells secreting related cytokines and the mRNA levels of transcription factors after stimulation with recombinant EBNA2 between the EBV-infected non-liver injury group and the infected liver injury group(P＞0.05).The levels of perforin secreted by CD8+T cells and granzyme B after stimulation with recombinant EBNA2 were higher in the EBV infection-induced liver injury group than those in the non-liver injury group[(75.51±23.33)pg/ml vs.(58.99±18.39)pg/ml,P=0.017][(117.8±44.55)pg/ml vs.(90.22±34.21)pg/ml,P=0.034].The mRNA levels of Fas ligand and tumor necrosis factor-related apoptosis-inducing ligand in CD8+T cells in the liver injury group caused by EBV infection were approximately 1.5 and 1.2 times higher than those in the non-liver injury group,respectively,and the difference was statistically significant(P＜0.001),but there was no statistically significant difference in the proportional expression of programmed cell death-1 and cytotoxic T lymphocyte-associated antigen-4 in CD8+T cells between the EBV-infected non-liver injury group and infected liver injury group(P＞0.05)Conclusion Patients with liver injury caused by EBV infection have strong virus-specific CD8+T cell toxic effects,which may mediate EBV-induced liver injury.
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