【摘要】：目的:探究腫瘤來源的Ig G是否對T細(xì)胞活化具有抑制作用。方法:首先從卵巢癌腫瘤組織純化了腫瘤來源的Ig G(tumor derived Ig G,t Ig G),并從臍帶血中分離出單個核細(xì)胞(cord blood mononuclear cells,CBMC)及淋巴細(xì)胞(cord blood lymphocyte,CBL),用植物血球凝集素(phytohaemagg lutinin,PHA)刺激單個核細(xì)胞和淋巴細(xì)胞,使單個核細(xì)胞與淋巴細(xì)胞處于增殖狀態(tài),用熒光染料羥基熒光素二醋酸鹽琥珀酰亞胺酯(carboxyfluorescein succinimidyl amino ester,CFSE)染色,CFSE對細(xì)胞無毒性,可以穿透細(xì)胞膜,在活細(xì)胞內(nèi)與胞內(nèi)蛋白結(jié)合,其熒光強(qiáng)度會隨著細(xì)胞的分裂增殖而逐級遞減,從而可以檢測細(xì)胞增殖的情況。把從卵巢癌腫瘤組織中純化得到的t Ig G分為1 mg/L、10 mg/L和100 mg/L組,分別加入到單個核細(xì)胞和淋巴細(xì)胞中,靜脈注射用免疫球蛋白(intravenous immunoglobulin,IVIG)作為陽性對照,也分為1 mg/L、10 mg/L和100 mg/L組,磷酸緩沖鹽溶液(phosphate buffer saline,PBS)作為陰性對照,用細(xì)胞流式術(shù)檢測CD4+及CD8+T細(xì)胞的增殖能力,并分別在64 h和86 h兩個時間點(diǎn)檢測細(xì)胞的增殖情況。結(jié)果:在CBMC體系中加入的t Ig G可以明顯地抑制CBMC中T細(xì)胞的增殖,發(fā)揮免疫抑制作用;同樣,在CBL體系中加入的t Ig G對T細(xì)胞也有抑制作用,且加入t Ig G刺激的T細(xì)胞比加入IVIG刺激的T細(xì)胞的抑制增殖作用更強(qiáng);在CBL體系中,t Ig G的1 mg/L組和10 mg/L組對T細(xì)胞發(fā)揮的抑制增殖作用比100 mg/L組弱。結(jié)論:t Ig G可以抑制T細(xì)胞的增殖,促使腫瘤發(fā)揮腫瘤免疫逃逸效應(yīng)。
[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.
【作者單位】： 北京大學(xué)基礎(chǔ)醫(yī)學(xué)院免疫學(xué)系;北京積水潭醫(yī)院婦產(chǎn)科;北京大學(xué)人民醫(yī)院婦產(chǎn)科;
【基金】：國家自然科學(xué)基金(81272237)資助~~
【分類號】：R730.3

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