CAJ | 학술논문

目的：研究卵巢癌患者腹水及外周血单个核细胞（PBMC）中CD4+CD25+调节性T细胞（Treg）的表达差异、免疫调节功能及其含量与化疗、复发的关系，探究其在卵巢癌腹腔微环境中发挥免疫调节的具体作用及意义。方法：采用流式细胞术分别检测27例卵巢癌患者腹水及28例卵巢癌患者PBMC中CD4+CD25+/CD4+T细胞百分比，并根据收集标本时患者临床特征进行分组，比较处于初治（PD）、化疗后（AC）及复发（RD）3个阶段卵巢癌患者腹水及PBMC中Treg含量的差异。应用免疫磁珠分选卵巢癌患者腹水及外周血Treg，与经羧基荧光素二醋酸盐琥珀酰亚胺酯（carboxyfluorescein succinimidylester，CFSE）标记的自体CD4+CD25-T细胞按不同比例（0：1，1：1，1：2及1：4）共培养，检测Treg免疫抑制功能。结果：卵巢癌患者腹水中CD4+CD25+/CD4+T细胞百分比（28.25±14.06）%较PBMC中（14.6±4.74）%显著增高（P<0.0001）。卵巢癌患者PD、AC及RD 3个阶段腹水及PBMC中Treg含量均显示为AC>RD>PD，且均有显著统计学意义（P<0.0001）。体外实验结果显示，卵巢癌患者腹水中CD4+CD25+Treg可有效抑制CFSE标记的自体CD4+CD25-T细胞增殖，且抑制功能较外周血显著增强（P<0.01）。结论：卵巢癌腹腔微环境中存在CD4+CD25+Treg，且其含量及免疫抑制功能较外周血PBMC显著增高，提示卵巢癌腹腔内更易发生免疫逃逸作用。卵巢癌患者化疗后及复发阶段腹水及PBMC中CD4+CD25+Treg含量均大于原发阶段，提示化疗可能促进卵巢癌患者体内Treg含量升高，而Treg升高可能参与促进肿瘤复发。
목적：연구란소암환자복수급외주혈단개핵세포（PBMC）중CD4+CD25+조절성T세포（Treg）적표체차이、면역조절공능급기함량여화료、복발적관계，탐구기재란소암복강미배경중발휘면역조절적구체작용급의의。방법：채용류식세포술분별검측27례란소암환자복수급28례란소암환자PBMC중CD4+CD25+/CD4+T세포백분비，병근거수집표본시환자림상특정진행분조，비교처우초치（PD）、화료후（AC）급복발（RD）3개계단란소암환자복수급PBMC중Treg함량적차이。응용면역자주분선란소암환자복수급외주혈Treg，여경최기형광소이작산염호박선아알지（carboxyfluorescein succinimidylester，CFSE）표기적자체CD4+CD25-T세포안불동비례（0：1，1：1，1：2급1：4）공배양，검측Treg면역억제공능。결과：란소암환자복수중CD4+CD25+/CD4+T세포백분비（28.25±14.06）%교PBMC중（14.6±4.74）%현저증고（P<0.0001）。란소암환자PD、AC급RD 3개계단복수급PBMC중Treg함량균현시위AC>RD>PD，차균유현저통계학의의（P<0.0001）。체외실험결과현시，란소암환자복수중CD4+CD25+Treg가유효억제CFSE표기적자체CD4+CD25-T세포증식，차억제공능교외주혈현저증강（P<0.01）。결론：란소암복강미배경중존재CD4+CD25+Treg，차기함량급면역억제공능교외주혈PBMC현저증고，제시란소암복강내경역발생면역도일작용。란소암환자화료후급복발계단복수급PBMC중CD4+CD25+Treg함량균대우원발계단，제시화료가능촉진란소암환자체내Treg함량승고，이Treg승고가능삼여촉진종류복발。
Objective:CD4+CD25+regulatory T cells (Treg) may contribute to tumor progression by suppressing antitumor im-munity. The function of Treg in antitumor immunity regulation in the peritoneal microenvironment of ovarian cancer (OC) was investi-gated and compared with the circulating Treg to elucidate OC immune escape. Methods: Flow cytometry was used to determine the proportion of CD4+CD25+T cells in CD4+T cells in ascites of 27 patients with OC and in peripheral blood lymphocytes of 28 patients with OC. The samples were analyzed and classified in three stages:primary disease (PD), after chemotherapy (AC), and recurrence dis-ease (RD), according to the clinical conditions of the OC patients upon donating the samples. The percentage of Treg in the three groups was determined in ascites and blood. CD4+CD25+T cells were isolated from ascites and peripheral blood of patients with OC us-ing magnetic sorting (MACS) system. The cells were then tested for regulatory function through coculture with carboxyfluorescein diac-etate succinimidyl ester-labeled autologous CD4+ CD25- responder cells. Results:The proportion of CD4+ CD25+T cells in CD4+T cells significantly increased in ascites (28.25%± 14.06%) compared with that in blood (14.6%± 4.74%;P<0.0001). The Treg in ascites and blood in AC showed higher proportion (P<0.0001) than those in the PD and RD;the proportion in RD was higher than that in PD (P<0.0001). Moreover, the Treg in ascites mediated a significantly higher suppression compared with the Treg in peripheral blood (P<0.001). Conclusion:The frequency and suppressor function of Treg were significantly higher in ascites than in peripheral blood. This finding suggests more possibility for escape immune surveillance in the peritoneal microenvironment. Moreover, the proportion of Treg in AC was higher than that in PD or RD;the proportion in RD was higher than that in the PD. Chemotherapy may favor the expansion of Treg, which may promote the recurrence of cancer.