Chiba M, Ohta H, Masamune O, Yoshida Y
Gastroenterol Jpn. 1986 Dec;21(6):584-93. doi: 10.1007/BF02774486.
Spontaneous cell-mediated cytotoxicity (SCMC) and the marker of natural killer (NK) cells mediating SCMC of the human large intestine were studied. Lamina proprial lymphoid cells (LPL) were isolated by sequential dithiothreitol-EDTA-collagenase treatment of the gut specimen. SCMC was measured by the chromium release method. Target cells included P4788 in monolayer, a cell line derived from colon cancer, Chang cells in monolayer, and K562 in suspension. Target cells in monolayer including colon cancer cell line were chosen because they were thought to be more appropriate to assess SCMC for lymphoid cells in the solid organ. While lower compared to cytotoxicities (CT) by peripheral blood lymphoid cells (PBL), define CT were observed in LPL against all three targets. NK cells marker was studied both on LPL by an indirect fluorescent antibody method and on the gut tissue by indirect immunoperoxidase staining using anti HNK-1 monoclonal antibody which defines virtually all NK cells. HNK-1 positive (HNK-1 +) cells were identified in both methods. HNK-1 + cells were observed in the epithelium, lamina propria, and lymph follicle with or without germinal centers. These results clearly demonstrated the presence of SCMC and HNK-1 + cells in the human large bowel.
对人结肠自发细胞介导的细胞毒性(SCMC)以及介导SCMC的自然杀伤(NK)细胞标志物进行了研究。通过对肠道标本依次进行二硫苏糖醇-乙二胺四乙酸-胶原酶处理来分离固有层淋巴细胞(LPL)。采用铬释放法测定SCMC。靶细胞包括单层培养的源自结肠癌的细胞系P4788、单层培养的Chang细胞以及悬浮培养的K562。选择包括结肠癌细胞系在内的单层培养靶细胞是因为它们被认为更适合评估实体器官中淋巴细胞的SCMC。虽然与外周血淋巴细胞(PBL)的细胞毒性(CT)相比更低,但在LPL中观察到针对所有三种靶细胞的明确CT。通过间接荧光抗体法在LPL上以及使用定义几乎所有NK细胞的抗HNK-1单克隆抗体通过间接免疫过氧化物酶染色在肠道组织上研究NK细胞标志物。在两种方法中均鉴定出HNK-1阳性(HNK-1 +)细胞。在有或无生发中心的上皮、固有层和淋巴滤泡中均观察到HNK-1 +细胞。这些结果清楚地证明了人结肠中存在SCMC和HNK-1 +细胞。