基于单细胞质谱流式技术分析膀胱肿瘤微环境中免疫细胞的组成

Analysis of the composition of immune cells in the bladder tumor microenvironment based on single-cell mass cytometry

目的探讨单细胞质谱流式技术(single-cell mass cytometry,CyTOF)分析膀胱癌组织与癌旁组织浸润性免疫细胞亚群特征的效果,为膀胱癌异质性研究提供新的策略。方法收集4例膀胱癌患者新鲜癌组织及其癌旁组织,通过酶解法联合机械法解离制备成单细胞悬液。采用CyTOF检测肿瘤浸润性免疫细胞中26种免疫相关标志物的表达情况。结果通过细胞相似性分析,将膀胱CD45^+免疫细胞分成25个亚群(CD4^+T细胞簇7个、CD8^+T细胞簇8个、巨噬细胞簇4个、B细胞簇2个、树突状细胞簇1个和其他类型的免疫细胞簇3个)。膀胱癌组织与癌旁组织浸润性免疫细胞亚群相对含量存在差异,膀胱癌组织中T细胞相对富集,占比为76.73%;癌旁组织中B细胞和巨噬细胞相对富集,占比分别为15.20%和23.82%。膀胱癌组织中CD4^+Treg细胞的平均占比高于癌旁组织(2.20%vs 0.81%,P=0.016)。结论CyTOF可有效区分膀胱肿瘤微环境中的免疫细胞亚群,实现对肿瘤异质性的检测,可为膀胱癌的精准治疗提供依据。

Objective To investigate the effect of single-cell mass cytometry(Cy TOF)analysis on the characteristics of infiltrating immune cell subsets in bladder cancer tissues and paracancerous tissues,providing a new strategy for the study of bladder cancer heterogeneity.Methods The cancer tissues and paracancerous tissues of 4 bladder cancer patients were collected and prepared into single-cell suspensions by enzymatic hydrolysis and mechanical dissociation.CyTOF was used to detect the expression of 26 immune-related markers in tumorinfiltrating immune cells.Results The bladder CD45^+immune cells were divided into 25 subgroups(7 CD4^+T cell clusters,8 CD8^+T cell clusters,4 macrophage clusters,2 B cell clusters,1 dendritic cell cluster,and 3 other types immune cell clusters)by cell similarity analysis.The contents of infiltrating immune cell subpopulations in bladder cancer tissues and paracancerous tissues were different.T cells were more abundant in bladder cancer tissues at 76.73%,whereas B cells and macrophages were more abundant in paracancerous tissues,accounting for 15.20%and 23.82%,respectively.The average proportion of CD4^+Treg cells in bladder cancer tissues was higher than that in paracancerous tissues(2.20%vs 0.81%,P=0.016).Conclusions CyTOF can effectively distinguish the immune cell subsets in the bladder tumor microenvironment and detect tumor heterogeneity,providing a basis for the accurate treatment of bladder cancer.