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具有针对肿瘤靶细胞的自然杀伤细胞活性的鸡肠道上皮内淋巴细胞的分离及功能特性研究

Isolation and functional characterization of chicken intestinal intra-epithelial lymphocytes showing natural killer cell activity against tumour target cells.

作者信息

Chai J Y, Lillehoj H S

机构信息

Protozoan Diseases Laboratory, U.S. Department of Agriculture, Beltsville, Maryland 20705.

出版信息

Immunology. 1988 Jan;63(1):111-7.

Abstract

Intestinal intra-epithelial lymphocytes (IEL) of SC or FP chickens were isolated and examined for their natural killer (NK)-cell activity against chicken tumour cell lines, LSCC-RP9 (RP9), LSCC-RP12 (RP12), MDCC-MSB-1 (MSB-1) and MDCC-CU36 (CU36). In general, IEL of satisfactory yield and of good viability were obtained with EDTA treatment of the gut tissues, followed by rapid passages of the resultant cells through nylon-wool columns and centrifugation on two-step Percoll density gradients (45% and 80%). In 4-hr and 16-hr 51Cr-release assays, the NK-cell activity of chicken IEL depended not only upon the type of target cells but also upon the incubation time and the host genetic background. RP9, MSB-1 and CU36 were susceptible to NK lysis by IEL and by spleen cells, while RP12 was resistant to lysis even after a prolonged incubation. In kinetic studies the cytotoxicity was detactable from 2 hr after incubation and progressively increased up to 16 or 18 hr. The IEL of SC chickens revealed significantly higher levels of NK-cell activity against RP9 than FP-strain chickens, whereas their splenic NK-cell activity was not significantly different. Against MSB-1 targets, however, IEL of SC and FP chickens showed similar levels of NK-cell activity while their spleens did not (being higher in FP). When tested in FP chickens, IEL NK-cell activity was inhibited by the addition of unlabelled homologous target cells. In general, NK-cell activity was higher in the jejunum and ileum than in the duodenum and caecum. Efforts to enrich IEL NK-effector cells by discontinuous Percoll gradients were not successful. The results of the present study show that IEL of chicken intestine contain effector cells that can mediate NK-cell activity against chicken tumour cells.

摘要

分离了SC或FP品系鸡的肠道上皮内淋巴细胞(IEL),并检测其对鸡肿瘤细胞系LSCC-RP9(RP9)、LSCC-RP12(RP12)、MDCC-MSB-1(MSB-1)和MDCC-CU36(CU36)的自然杀伤(NK)细胞活性。一般来说,通过用EDTA处理肠道组织,然后将所得细胞快速通过尼龙毛柱并在两步Percoll密度梯度(45%和80%)上离心,可获得产量令人满意且活力良好的IEL。在4小时和16小时的51Cr释放试验中,鸡IEL的NK细胞活性不仅取决于靶细胞的类型,还取决于孵育时间和宿主遗传背景。RP9、MSB-1和CU36易被IEL和脾细胞进行NK裂解,而RP12即使经过长时间孵育也对裂解具有抗性。在动力学研究中,孵育2小时后即可检测到细胞毒性,并逐渐增加至16或18小时。SC品系鸡的IEL对RP9显示出比FP品系鸡显著更高水平的NK细胞活性,而它们的脾NK细胞活性没有显著差异。然而,对于MSB-1靶细胞,SC和FP品系鸡的IEL显示出相似水平的NK细胞活性,而它们的脾脏则不然(FP品系鸡的脾脏活性更高)。在用FP品系鸡进行测试时,加入未标记的同源靶细胞会抑制IEL的NK细胞活性。一般来说,空肠和回肠中的NK细胞活性高于十二指肠和盲肠。通过不连续Percoll梯度富集IEL NK效应细胞的努力未成功。本研究结果表明,鸡肠道的IEL含有可介导针对鸡肿瘤细胞的NK细胞活性的效应细胞。

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