肿瘤相关中性粒细胞在胃癌中的分布、表型及其免疫抑制功能研究

Distribution,phenotype and immunosuppressive function of tumor-associated neutrophils in gastric cancer

目的分析胃癌患者外周血中及不同类型胃组织中的中性粒细胞的分布情况,并研究其表型,探讨胃癌组织培养上清诱导的条件性中性粒细胞对CD3+T细胞功能的影响。方法采用流式细胞术检测并分析42例胃癌患者的胃癌组织、癌旁组织、正常胃组织及外周血中CD66b+中性粒细胞的比例及表型,同时采用免疫组织化学染色技术针对中性粒细胞表面标志CD15原位分析不同类型胃组织石蜡切片中中性粒细胞的浸润情况,最后用胃癌组织培养上清诱导的条件性中性粒细胞与CD3+T细胞共培养,检测其对T细胞增殖能力的影响。结果胃癌患者外周血中性粒细胞比例明显高于健康成人(P<0.01),胃癌组织中的中性粒细胞的频率也高于癌旁组织(P<0.05)和正常胃组织(P<0.05);在胃癌患者外周血及不同类型胃组织浸润的中性粒细胞可能主要为成熟的分叶核粒细胞;相较于正常胃组织,胃癌组织培养上清诱导的条件性中性粒细胞能显著抑制CD3+T细胞增殖(P<0.01)。结论中性粒细胞在胃癌微环境中浸润增高,其可能通过抑制CD3+T细胞增殖而促进胃癌进展。

Objective To analyze the distribution and phenotype of neutrophils in periphery blood and different types of gastric tissues from gastric cancer （GC） patients, and to investigate the influence of GC tissue culture supernatants induced-conditioned neutrophils （GCSI-CNs） on CD3 ＋ T cells. Methods Flow cytome-try was employed to detect and analyze the ratio and phenotype of CD66b ＋ neutrophils in tumoral, peritumoral and normal gastric tissues and peripheral blood of 42 GC patients. Then, paraffin-embedded tissues of GC patients were investigated immunohistochemically with anti-human CD15 antibody to analyze the infiltration of neutrophils in situ. Finally, GCSI-CNs were cocuhured with CD3 ＋ T cells, and the proliferation of CD3 ＋ T cells was analyzed by flow cytometry. Results The percentage of neutrophils in periphery blood of GC patients was much higher than that in healthy donors （P 〈 0. 01 ）. The frequency of neutrophils was significantly higher in tumoral gastric tissues than in peritumoral （P 〈 0. 05 ） or in normal gastric tissues （P 〈 0. 05 ）. Meanwhile, neutrophils infiltrated in different types of gastric tissues and in periphery blood of GC patients were probably mainly mature segmented neutrophils. In addition, GCSI-CNs significantly inhibited the proliferation of CD3 ＋ T cells. Conclusion Neutrophils are accumulated in GC microenvironment and may facilitate the progression of GC by inhibiting the proliferation of CD3 ＋ T cells.