Ⅲ-Ⅳa期鼻咽癌LMP特异性T细胞免疫反应与临床分析

Analysis of LMP specific T cell immune response and clinical factors in stage III-IVa nasopharyngeal carcinoma

目的探讨Ⅲ-Ⅳa期鼻咽癌EB病毒潜伏膜蛋白(LMP)1、LMP2特异性T细胞免疫反应及临床意义,为鼻咽癌免疫治疗提供思路和依据。方法收集新疆医科大学附属肿瘤医院2018年2月至2020年10月初治的59例鼻咽癌患者,应用LMP抗原刺激外周血单核细胞,胞内细胞因子染色及流式细胞术研究CD4^(+)T细胞、CD8^(+)T细胞白细胞介素(IL)-2、IL-13、干扰素γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)4种细胞因子的表达,并结合临床资料分析。结果鼻咽癌患者外周血T细胞对LMP1、LMP2反应阳性率存在差异。T_(3)-T_(4)期鼻咽癌患者LMP1特异性CD4^(+)T细胞阳性率明显高于T_(1)-T_(2)期患者(51.0%∶10.0%,P=0.042),LMP1和LMP2、CD4^(+)T细胞和CD8^(+)T细胞间部分细胞因子表达也存在差异。生存分析显示,2、3年OS率为91.5%、88.2%,2、3年PFS率为83.3%、75.3%;单因素分析提示吸烟史、男性、LMP1刺激CD4^(+)T细胞IL-13阳性表达是疾病的进展的危险因素(P=0.026、0.045、0.006);多因素分析显示LMP1刺激CD4^(+)T细胞IL-13阳性表达与吸烟史是局部晚期鼻咽癌患者PFS的独立危险因素(P=0.017、0.019)。结论LMP在鼻咽癌患者外周血中产生特异性T细胞免疫反应,不同T细胞亚群表达存在差异。LMP1、LMP2特异性T细胞免疫反应与原发肿瘤大小、转移淋巴结体积有关。LMP1刺激CD4^(+)T细胞IL-13阳性表达和吸烟史影响疾病的进展。

Objective To explore the Epstein-Barr virus(EBV)latent infection membrane protein(LMP)1 or LMP2 specific T cell immune response and clinical significance in stage III-IVa nasopharyngeal carcinoma(NPC),aiming to provide ideas and evidence for immunotherapy in NPC.Methods Fifty-nine NPC patients admitted to the Affiliated Tumor Hospital of Xinjiang Medical University from February 2018 to October 2020 for primary treatment were collected.Peripheral blood monocytes(PBMCs)were stimulated by LMP antigen.Intracellular cytokine staining and flow cytometry were applied to study the expression levels of IL-2,IL-13,interferon-γ(IFN-γ)and tumor necrosis factor-α(TNF-α)from CD4^(+)T and CD8^(+)T cells,and then analyzed in conjunction with clinical factors.Results The positive rates of total PBMCs to LMP1 and LMP2 in NPC patients were different.The positive rate of LMP1 specific CD4^(+)T cells was statistically higher in stage T_(3)-T_(4) NPC than that in stage T_(1)-T_(2)(51.0%vs.10.0%,P=0.042).There were also differences in the expression of cytokines between LMP1 and LMP2,CD4^(+)T cells and CD8^(+)T cells.Survival analysis showed the 2-year and 3-year overall survival(OS)rates were 91.5%and 88.2%,and the 2-year and 3-year progression-free survival(PFS)rates were 83.3%and 75.3%.Univariate analysis suggested that smoking history,male and LMP1 stimulated IL-13 positive expression in CD4^(+)T cells affected the disease progression(P=0.026,0.045 and 0.006);multivariate analysis showed LMP1 stimulated IL-13 positive expression in CD4^(+)T cells and smoking history were the independent prognostic factors affecting PFS(P=0.017,0.019).Conclusions LMP1 and LMP2 generate specific T-cell immune response in PBMCs of NPC patients,with differential expression in two T-cell subsets.LMP1 and LMP2 specific T cell immune response is associated with primary tumor size and metastatic lymph node volume.LMP1 stimulated IL-13 positive expression in CD4^(+)T cells and smoking history affects the disease progression.