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目的：评价食管癌患者外周血恶性黑素瘤相关抗原（melanoma-associated antigen, MAGE)-C2抗原特异性CD8+T细胞的功能，并分析其与临床病理参数的相关性。方法：密度梯度离心法获取130例食管癌初治患者(收集自2014年11月至2015年11月郑州大学第一附属医院胸外科）的PBMC，流式细胞术、RT-PCR方法筛选HLA-A2*0201和MAGE-C2共阳性的患者，其PBMC培养7、14 d后加MAGE-C2(336-344)抗原肽1 μg/ml、IL-2 100 μg/ml 和CD3/CD28 Dynabeads 2 μl处理，诱导扩增抗原特异性CD8+ T细胞，流式细胞术检测CD8+ T细胞表面CD107a和胞内IFN-γ 的水平，并分析其与临床参数的相关性。结果：筛选出52例HLA-A2*0201和MAGE-C2共阳性的患者，MAGE-C2(336-344)处理其PBMC后，MAGE-C2(336-344)处理组CD8+T细胞中CD107a+IFN-γ+,CD107a+IFN-γ-和CD107a-IFN-γ+细胞的比例明显升高（P<0.05）；CD107a的表达在早期、未发生淋巴结转移和高分化的食管癌患者明显高于晚期、发生淋巴结转移和低分化的癌症患者（P<0.05）；IFN-γ 的表达在高分化的食管癌患者明显高于低分化的食管癌患者（P<0.05）。结论：成功诱导扩增MAGE-C2抗原特异性CD8+ T细胞，并发现其功能与食管癌的分期和淋巴结转移相关，似可作为一种理想的免疫治疗靶点。
OBJECTIVE: To evaluate the function of melanoma-associated antigen (MAGE) -C2 antigen-specific CD8 + T cells in peripheral blood of patients with esophageal cancer and analyze its correlation with clinicopathological parameters. Methods: PBMC from 130 patients with newly diagnosed esophageal cancer (collected from the Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University from November 2014 to November 2015) were obtained by density gradient centrifugation. The expression of HLA- A2 * 0201 and MAGE-C2 were co-positive. PBMCs were cultured for 7 and 14 days and then added with 1 μg / ml MAGE-C2 (336-344) antigen peptide, 100 μg / ml IL- The antigen-specific CD8 + T cells were induced to proliferate. The levels of CD107a and IFN-γ on the surface of CD8 + T cells were detected by flow cytometry. The correlations were analyzed with clinical parameters. Results: Fifty-two patients with positive co-positive HLA-A2 * 0201 and MAGE-C2 were screened for CD107a + CD8 + T cells in MAGE-C2 (336-344) The proportion of IFN-γ +, CD107a + IFN-γ- and CD107a-IFN-γ + cells was significantly increased (P <0.05). The expression of CD107a was significantly higher in patients with early stage esophageal cancer without lymph node metastasis and well- In advanced stage, patients with lymph node metastasis and poorly differentiated cancer (P <0.05); IFN-γ expression was significantly higher in poorly differentiated esophageal cancer patients with well-differentiated esophageal cancer (P <0.05). Conclusion: The successful induction of MAGE-C2 antigen-specific CD8 + T cells, and found that its function and esophageal cancer staging and lymph node metastasis may seem as an ideal target for immunotherapy.