肿瘤来源的IgG抑制脐带血中T细胞的增殖
发布时间：2018-10-10 19:05
【摘要】：目的:探究肿瘤来源的Ig G是否对T细胞活化具有抑制作用。方法:首先从卵巢癌肿瘤组织纯化了肿瘤来源的Ig G(tumor derived Ig G,t Ig G),并从脐带血中分离出单个核细胞(cord blood mononuclear cells,CBMC)及淋巴细胞(cord blood lymphocyte,CBL),用植物血球凝集素(phytohaemagg lutinin,PHA)刺激单个核细胞和淋巴细胞,使单个核细胞与淋巴细胞处于增殖状态,用荧光染料羟基荧光素二醋酸盐琥珀酰亚胺酯(carboxyfluorescein succinimidyl amino ester,CFSE)染色,CFSE对细胞无毒性,可以穿透细胞膜,在活细胞内与胞内蛋白结合,其荧光强度会随着细胞的分裂增殖而逐级递减,从而可以检测细胞增殖的情况。把从卵巢癌肿瘤组织中纯化得到的t Ig G分为1 mg/L、10 mg/L和100 mg/L组,分别加入到单个核细胞和淋巴细胞中,静脉注射用免疫球蛋白(intravenous immunoglobulin,IVIG)作为阳性对照,也分为1 mg/L、10 mg/L和100 mg/L组,磷酸缓冲盐溶液(phosphate buffer saline,PBS)作为阴性对照,用细胞流式术检测CD4+及CD8+T细胞的增殖能力,并分别在64 h和86 h两个时间点检测细胞的增殖情况。结果:在CBMC体系中加入的t Ig G可以明显地抑制CBMC中T细胞的增殖,发挥免疫抑制作用;同样,在CBL体系中加入的t Ig G对T细胞也有抑制作用,且加入t Ig G刺激的T细胞比加入IVIG刺激的T细胞的抑制增殖作用更强;在CBL体系中,t Ig G的1 mg/L组和10 mg/L组对T细胞发挥的抑制增殖作用比100 mg/L组弱。结论:t Ig G可以抑制T细胞的增殖,促使肿瘤发挥肿瘤免疫逃逸效应。
[Abstract]:Objective: to investigate whether tumor derived Ig G can inhibit T cell activation. Methods: Ig G (tumor derived Ig Glut Ig G), was purified from ovarian cancer tissue and mononuclear cells (cord blood mononuclear cells,CBMC) and lymphocytes (cord blood lymphocyte,CBL) were isolated from umbilical cord blood. Mononuclear cells and lymphocytes were stimulated with phytohemagglutinin (phytohaemagg lutinin,PHA). When mononuclear cells and lymphocytes were proliferated and stained with fluorescent dye hydroxyfluorescein diacetate succinimide (carboxyfluorescein succinimidyl amino ester,CFSE), CFSE was nontoxic to cells and could penetrate the cell membrane and bind to intracellular proteins in living cells. The fluorescence intensity decreases with cell division and proliferation, which can detect cell proliferation. T Ig G purified from ovarian cancer tissue was divided into 1 mg/L,10 mg/L group and 100 mg/L group, which were added to mononuclear cells and lymphocytes, respectively. Immunoglobulin (intravenous immunoglobulin,IVIG) was used as positive control for intravenous injection, and was also divided into 1 mg/L,10 mg/L and 100 mg/L groups. Phosphoric acid buffer solution (phosphate buffer saline,PBS) was used as negative control. The proliferation of CD4 and CD8 T cells was detected by flow cytometry, and the proliferation of CD4 and CD8 T cells was detected at 64 h and 86 h, respectively. Results: the addition of t Ig G in CBMC system could obviously inhibit the proliferation of T cells in CBMC and play an immunosuppressive role, and the addition of t Ig G in CBL system could also inhibit the proliferation of T cells. The inhibitory effect of T cells stimulated by t Ig G was stronger than that of T cells stimulated by IVIG, and the inhibitory effect of 1 mg/L group and 10 mg/L group of, t Ig G on T cell proliferation was weaker than that of 100 mg/L group in CBL system. Conclusion: t Ig G can inhibit T cell proliferation and promote tumor immune escape.
【作者单位】： 北京大学基础医学院免疫学系;北京积水潭医院妇产科;北京大学人民医院妇产科;
【基金】：国家自然科学基金(81272237)资助~~
【分类号】：R730.3
本文编号：2262908
[Abstract]:Objective: to investigate whether tumor derived Ig G can inhibit T cell activation. Methods: Ig G (tumor derived Ig Glut Ig G), was purified from ovarian cancer tissue and mononuclear cells (cord blood mononuclear cells,CBMC) and lymphocytes (cord blood lymphocyte,CBL) were isolated from umbilical cord blood. Mononuclear cells and lymphocytes were stimulated with phytohemagglutinin (phytohaemagg lutinin,PHA). When mononuclear cells and lymphocytes were proliferated and stained with fluorescent dye hydroxyfluorescein diacetate succinimide (carboxyfluorescein succinimidyl amino ester,CFSE), CFSE was nontoxic to cells and could penetrate the cell membrane and bind to intracellular proteins in living cells. The fluorescence intensity decreases with cell division and proliferation, which can detect cell proliferation. T Ig G purified from ovarian cancer tissue was divided into 1 mg/L,10 mg/L group and 100 mg/L group, which were added to mononuclear cells and lymphocytes, respectively. Immunoglobulin (intravenous immunoglobulin,IVIG) was used as positive control for intravenous injection, and was also divided into 1 mg/L,10 mg/L and 100 mg/L groups. Phosphoric acid buffer solution (phosphate buffer saline,PBS) was used as negative control. The proliferation of CD4 and CD8 T cells was detected by flow cytometry, and the proliferation of CD4 and CD8 T cells was detected at 64 h and 86 h, respectively. Results: the addition of t Ig G in CBMC system could obviously inhibit the proliferation of T cells in CBMC and play an immunosuppressive role, and the addition of t Ig G in CBL system could also inhibit the proliferation of T cells. The inhibitory effect of T cells stimulated by t Ig G was stronger than that of T cells stimulated by IVIG, and the inhibitory effect of 1 mg/L group and 10 mg/L group of, t Ig G on T cell proliferation was weaker than that of 100 mg/L group in CBL system. Conclusion: t Ig G can inhibit T cell proliferation and promote tumor immune escape.
【作者单位】： 北京大学基础医学院免疫学系;北京积水潭医院妇产科;北京大学人民医院妇产科;
【基金】：国家自然科学基金(81272237)资助~~
【分类号】：R730.3
【相似文献】
相关期刊论文 前4条
1 王锴佳;唐建华;;肿瘤来源的外体及其在肿瘤中的作用[J];中南大学学报(医学版);2010年12期
2 孙继军,姜广水,孙善珍;肿瘤来源的外体的研究进展[J];山东大学基础医学院学报;2005年01期
3 王春梅;;肿瘤来源的COX-2诱导MSC精氨酸酶Ⅰ的产生[J];中国肿瘤生物治疗杂志;2007年01期
4 ;循环中肿瘤来源膜片段与子宫内膜癌的分期有关[J];中华医学信息导报;2002年10期
，本文编号：2262908
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2262908.html
