目的:研究淋巴细胞亚群比例在食管癌患者肿瘤引流淋巴结(tumor—draininglymphnode,TDLN)中的变化及其在肿瘤进展中的意义。方法:收集在河北医科大学第四医院行食管癌切除术患者的引流淋巴结样本70例,根据转移情况将样本分为转移组和未转移组,并根据患者肿瘤临床分期标准分为早期组和晚期组,无菌分离TDLN内淋巴细胞并应用流式细胞术检测以下亚群在总淋巴细胞中所占比例:CD3+T细胞、CD3+CD4+T细胞、CD3+CD8+T细胞、CD3-CD19+B细胞、CD3-CDl6+CD56+NK细胞和CD4+CD25+Treg细胞。采用Pearson积差相关分析Treg细胞与其他淋巴细胞比例之间的关系,采用t检验或Mann—WhitneyU检验分析各组间TDLN淋巴细胞亚群的差异。结果:食管癌患者TDLN中的Treg细胞与T细胞、CIM+T细胞比例显著相关(P=0.002,P=0.003),与CD8+T细胞、B细胞、NK细胞比例无相关性(P〉0.05)。与未转移组淋巴结相比,转移组的T细胞、CD4+T细胞、CD8+T细胞和NK细胞比例显著降低(P=0.000,P=0.000,P=0.016,P=0.038),B细胞、Treg细胞比例显著提高(P=0.000,P=0.018),CD4+/CD8+T细胞比值差异没有统计学意义(P=0.687)。与早期食管癌组淋巴结相比,晚期食管癌组的T细胞、CD4+T细胞、CD8+T细胞和NK细胞比例显著降低(P=0.000,P=0.008,P=0.027,P=0.022),B细胞、Treg细胞比例显著提高(P=0.000,P=0.043),CD4+/CD8+T细胞比值差异没有统计学意义(P=0.770)。结论:食管癌患者TDLN内淋巴细胞亚群分布紊乱,是促进肿瘤向淋巴结转移并提高临床分期的重要因素之一。
Objective: To investigate changes in lymphocyte subsets in tumor-draining lymph nodes (TDLNs) of esophageal cancer patients and in association with tumor progression. Methods: A total of 70 TDLN specimens were col- lected from esophageal cancer patients who underwent esophagectomy in the Fourth Hospital of Hebei Medical University. They were categorized as metastatic and non-metastatic based on their metastatic statuses, and as early stage and advanced stage according the TNM staging. Lymphocytes were isolated sterilely from these TDLN specimens. Proportions of CD3 + T cell, CD3 + CD4 + T cell, CD3 + CD8 + T cell, CD3 - CD19 + B cell, CD3 - CD16 + CD56 + NK cell and CD4 + CD25 + Treg in the mixed lymphocyte preparations were analyzed by flow cytometry. The relationships between Treg and other lympho- cyte subsets were examined by Pearson' s correlation test. The differences in lymphocyte subset proportions were analyzed by student t test or Mann-Whitney U test. Results: The proportion of Treg was correlated with proportions of T cells ( P = 0. 002) and CD4 + T cells (P = 0. 003 ) but not with proportions of CD8 + T cells, B ceils and NK cells ( P 〉 0.05 ). Compared to the non-metastatic group, proportions of T ceils ( P 〈 0. 001 ), CD4 + T cells ( P 〈 0. 001 ), CD8 + T cells(P =0. 016) and NK cells (P =0.038) in the metastatic group were significantly decreased, while proportions of B cells (P 〈0.001 ) and Treg (P =0.018) were significantly increased and the CD4+/CD8 + T cell ratio was not significant differ- ent (P =0.687). Proportions of T cells (P 〈0.001), CD4 + T cells (P =0.008), CD8 + T cells (P =0.027) and NK cells ( P = 0.022) were significantly lower specimens but B cell (P 〈 0.001 ) and Treg ( P = 0.043 ) proportions were significantly higher in the advanced stage specimens than in the early stage specimens, while difference in the CD4+/CD8 + T cell ratio was not statistically