CAJ | 학술논문

目的：研究卵巢癌患者腹水内CD4+CD25+调节性T细胞（Treg）免疫抑制功能与患者临床病理特点的关系及其与患者初治及复发状态是否相关，并进一步探讨卵巢癌腹水内CD4+CD25+调节性T细胞（Treg）发挥免疫调节性作用的具体机制。方法：应用免疫磁珠分选法分选28例卵巢癌患者腹水内CD4+CD25+调节性T细胞及CD4+CD25-T细胞，采用羧基荧光素二醋酸盐琥珀酰亚胺酯（carboxyfluorescein succinimidylester，CFSE）标记CD4+CD25-T细胞，CD4+CD25+Treg与自体CFSE标记CD4+CD25-T细胞以1∶1比例共培养，经流式检测CFSE表达及Modfit软件分析CD4+CD25-T增殖指数（PI），计算各标本内Treg对自体CD4+CD25-T细胞增殖抑制率。应用中合性抗IL-10抗体及中合性抗TGF-β1抗体探究腹水内CD4+CD25+Treg介导免疫逃逸作用是否通过抑制性细胞因子IL-10或TGF-β1发挥作用。结果：Ⅲ～Ⅳ期卵巢癌腹水内CD4+CD25+Treg对自体CD4+CD25-T细胞增殖抑制率为（75.72±17.04）%，较Ⅰ～Ⅱ期Treg抑制率（59.61±16.97）%显著增强（P<0.05）。复发卵巢癌患者腹水内Treg对自体CD4+CD25-T细胞增殖抑制率为（85.89±7.07）%，较初治卵巢癌患者腹水Treg抑制率（52.82±8.18）%显著增强（P=0.0001）。共培养体系内加入中合性抗IL-10抗体或/及中和性抗TGF-β1抗体，Treg对自体CD4+CD25-T细胞增殖抑制率较对照组均显著降低（P<0.05）。结论：卵巢癌腹水内CD4+CD25+Treg介导免疫逃逸能力与肿瘤分期及复发、初治状态相关，且发挥免疫逃逸作用可能是与分泌抑制性细胞因子IL-10及TGF-β1有关。
목적：연구란소암환자복수내CD4+CD25+조절성T세포（Treg）면역억제공능여환자림상병리특점적관계급기여환자초치급복발상태시부상관，병진일보탐토란소암복수내CD4+CD25+조절성T세포（Treg）발휘면역조절성작용적구체궤제。방법：응용면역자주분선법분선28례란소암환자복수내CD4+CD25+조절성T세포급CD4+CD25-T세포，채용최기형광소이작산염호박선아알지（carboxyfluorescein succinimidylester，CFSE）표기CD4+CD25-T세포，CD4+CD25+Treg여자체CFSE표기CD4+CD25-T세포이1∶1비례공배양，경류식검측CFSE표체급Modfit연건분석CD4+CD25-T증식지수（PI），계산각표본내Treg대자체CD4+CD25-T세포증식억제솔。응용중합성항IL-10항체급중합성항TGF-β1항체탐구복수내CD4+CD25+Treg개도면역도일작용시부통과억제성세포인자IL-10혹TGF-β1발휘작용。결과：Ⅲ～Ⅳ기란소암복수내CD4+CD25+Treg대자체CD4+CD25-T세포증식억제솔위（75.72±17.04）%，교Ⅰ～Ⅱ기Treg억제솔（59.61±16.97）%현저증강（P<0.05）。복발란소암환자복수내Treg대자체CD4+CD25-T세포증식억제솔위（85.89±7.07）%，교초치란소암환자복수Treg억제솔（52.82±8.18）%현저증강（P=0.0001）。공배양체계내가입중합성항IL-10항체혹/급중화성항TGF-β1항체，Treg대자체CD4+CD25-T세포증식억제솔교대조조균현저강저（P<0.05）。결론：란소암복수내CD4+CD25+Treg개도면역도일능력여종류분기급복발、초치상태상관，차발휘면역도일작용가능시여분비억제성세포인자IL-10급TGF-β1유관。
Objective: This research explores the relationship between the immuno-suppression function of regulatory T cells (Treg) in the ascites of ovarian cancer (OC) patients, the clinico-pathologic features of these patients, and the correlation of the function of Treg with initial treatment and relapse status of the patients to further investigate the specific mechanism of immuno-regulatory func-tion of CD4+ CD25+ Treg in the ascites of OC. Methods: Immuno-magnetic activated cell sorting (MACS) was conducted to sort CD4+CD25+Treg and autologous CD4+CD25-Treg from the ascites of 28 OC patients. Carboxyfluorescein-diacetate succinimidyl ester (CFSE) was used to label the autologous CD4+CD25-Treg. These labeled cells were then used as controls and co-cultured with autologous CD4+CD25+Treg at the ratio of 1∶1 or 1∶2. The mean inhibition ratio of Treg in specimens to the proliferation of autolo-gous CD4+ CD25-Treg was calculated after the flow cytometry of the CFSE expression and Modfit software analysis of the CD4+CD25-Treg proliferation index (PI) were performed. Anti-IL-10 and/or anti-TGF-β1 antibodies were neutralized to investigate whether the CD4+CD25+Treg-mediated immuno-suppression escaped through the ascites can produce a marked effect by the inhibitory cyto-kine IL-10 or TGF-β1. Results: The mean inhibition ratio of CD4+ CD25- Treg in the ascites of stage Ⅲ to Ⅳ OC patients was (75.72±17.04)%, which is significantly higher than that of stageⅠtoⅡOC patients (59.61±16.97)%;P<0.05. In addition, Treg in the as-cites of OC patients with recurrent disease showed a significantly higher inhibition ratio than that of patients with primary disease;P<0.001. Moreover, Treg in groups added into neutralizing anti-IL-10 and/or anti-TGF-β1 antibodies displayed significantly lower depres-sant effect than the control group;P<0.05. Conclusion:The immuno-suppression of CD4+CD25+Treg in the ascites of OC patients is correlated with the tumor staging and status of the primary or recurrent diseases. Moreover, Treg may indicate a suppressor function by secreting cytokine IL-10 and TGF-β1.