Functional analysis of virus-specific CD4 +T cells and CD8 +T cells in patients with liver injury caused by Epstein-Barr virus infection
10.3760/cma.j.cn501113-20231122-00214
- VernacularTitle:EB病毒感染所致肝脏损伤患者病毒特异性CD4 +T细胞和CD8 +T细胞功能分析
- Author:
Yu LI
1
;
Fengyu XI
;
Weihua ZHANG
;
Ying GAO
Author Information
1. 陕西省人民医院感染性疾病科，西安 710068
- Keywords:
Liver injury;
Epstein-Barr virus;
CD4 + T lymphocytes;
CD8 + T lymphocytes
- From:
Chinese Journal of Hepatology
2024;32(1):64-71
- CountryChina
- Language:Chinese
-
Abstract:
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.
