中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2015年
4期
241-244
,共4页
邵立虹%金霖霖%左斯尧%魏巍%董卓%张海芹%韩海玲%高辉%金顺子
邵立虹%金霖霖%左斯堯%魏巍%董卓%張海芹%韓海玲%高輝%金順子
소립홍%금림림%좌사요%위외%동탁%장해근%한해령%고휘%금순자
电离辐射%A549细胞系%肿瘤微环境%Treg细胞%免疫耐受
電離輻射%A549細胞繫%腫瘤微環境%Treg細胞%免疫耐受
전리복사%A549세포계%종류미배경%Treg세포%면역내수
Radiation%A549 cell line%Tumor microenvironment%Regulatory T cells%Immune tolerance
目的 检测照射后与肺腺癌A549细胞共培养的淋巴细胞中调节性T细胞(Treg细胞)及其相关细胞因子和共刺激分子表达的变化,探讨辐射对肿瘤微环境中免疫耐受机制的影响.方法 分离人外周血淋巴细胞单独培养或与肺腺癌A549细胞进行共培养一段时间后,以6GyX射线进行照射,继续共培养一段时间后,以流式细胞术检测淋巴细胞中CD4+ CD25+、CD4+ CD25+FoxP3+、CD4+ CD25+ NrP1+不同亚组的百分数、共刺激分子ICOS和CTLA-4的表达情况;ELISA法检测培养体系上清中IL-10、TGF-β、IL-17的分泌量.结果 与照射前后单独培养组相比,相应条件下共培养组中CD4+ CD25+、CD4+ CD25+ FoxP3+、CD4+ CD25+ NrP1+、ICOS、CTLA-4、IL-10和TGF-β比例均增加(t=2.78 ～40.61,P＜0.05).而与照射前相比,照射后单独培养组和共培养组的CD4+ CD25+和CTLA-4比例增加(t=2.96、2.94、4.47、2.77,P＜0.05).细胞因子IL-17的表达量在肺腺癌A549细胞和X射线分别作用下无明显变化,但共同作用后会明显增加(t=4.00、4.71,P ＜0.05).结论 辐射刺激肺癌细胞肿瘤微环境中的CD4+ CD25+Treg细胞亚组及其相关CTLA-4和IL-17的表达,诱导肿瘤微环境中的免疫耐受作用.
目的 檢測照射後與肺腺癌A549細胞共培養的淋巴細胞中調節性T細胞(Treg細胞)及其相關細胞因子和共刺激分子錶達的變化,探討輻射對腫瘤微環境中免疫耐受機製的影響.方法 分離人外週血淋巴細胞單獨培養或與肺腺癌A549細胞進行共培養一段時間後,以6GyX射線進行照射,繼續共培養一段時間後,以流式細胞術檢測淋巴細胞中CD4+ CD25+、CD4+ CD25+FoxP3+、CD4+ CD25+ NrP1+不同亞組的百分數、共刺激分子ICOS和CTLA-4的錶達情況;ELISA法檢測培養體繫上清中IL-10、TGF-β、IL-17的分泌量.結果 與照射前後單獨培養組相比,相應條件下共培養組中CD4+ CD25+、CD4+ CD25+ FoxP3+、CD4+ CD25+ NrP1+、ICOS、CTLA-4、IL-10和TGF-β比例均增加(t=2.78 ～40.61,P＜0.05).而與照射前相比,照射後單獨培養組和共培養組的CD4+ CD25+和CTLA-4比例增加(t=2.96、2.94、4.47、2.77,P＜0.05).細胞因子IL-17的錶達量在肺腺癌A549細胞和X射線分彆作用下無明顯變化,但共同作用後會明顯增加(t=4.00、4.71,P ＜0.05).結論 輻射刺激肺癌細胞腫瘤微環境中的CD4+ CD25+Treg細胞亞組及其相關CTLA-4和IL-17的錶達,誘導腫瘤微環境中的免疫耐受作用.
목적 검측조사후여폐선암A549세포공배양적림파세포중조절성T세포(Treg세포)급기상관세포인자화공자격분자표체적변화,탐토복사대종류미배경중면역내수궤제적영향.방법 분리인외주혈림파세포단독배양혹여폐선암A549세포진행공배양일단시간후,이6GyX사선진행조사,계속공배양일단시간후,이류식세포술검측림파세포중CD4+ CD25+、CD4+ CD25+FoxP3+、CD4+ CD25+ NrP1+불동아조적백분수、공자격분자ICOS화CTLA-4적표체정황;ELISA법검측배양체계상청중IL-10、TGF-β、IL-17적분비량.결과 여조사전후단독배양조상비,상응조건하공배양조중CD4+ CD25+、CD4+ CD25+ FoxP3+、CD4+ CD25+ NrP1+、ICOS、CTLA-4、IL-10화TGF-β비례균증가(t=2.78 ～40.61,P＜0.05).이여조사전상비,조사후단독배양조화공배양조적CD4+ CD25+화CTLA-4비례증가(t=2.96、2.94、4.47、2.77,P＜0.05).세포인자IL-17적표체량재폐선암A549세포화X사선분별작용하무명현변화,단공동작용후회명현증가(t=4.00、4.71,P ＜0.05).결론 복사자격폐암세포종류미배경중적CD4+ CD25+Treg세포아조급기상관CTLA-4화IL-17적표체,유도종류미배경중적면역내수작용.
Objective To explore the immune tolerance in tumor microenvironment,we detected the expressions of CD receptors and related cytokines in Treg cells after being co-cultured with irradiated lung cancer cells.Methods Lymphocytes were isolated from human peripheral blood and cultured with lung adenocarcinoma cells A549,after irradiation with 6 Gy X-rays,the expression of CD4 + CD25 +,CD4 + CD25 + FoxP3+,CD4 + CD25 + NrP1+ and ICOS,CTLA-4 in the lymphocytes were tested by flow cytometry,and the contents of IL-10,TGF-β and IL-17 in the supernatant were detected by ELISA.Results Compared with lymphocytes alone culture,the expressions of CD4 + CD25 +,CD4 + CD25 + FoxP3+,CD4 + CD25 + NrP1 +,ICOS,CTLA-4,IL-10 and TGF-β in the co-culture groups with and without irradiation were all increased (t =2.78-40.61,P ＜ 0.05),the CD4 + CD25 + and CTLA-4 in the lymphocytes were increased after irradiation (t =2.96,2.94,4.47,2.77,P ＜ 0.05).The content of IL-17 in the supernant were increased after co-cuture with irradiated A549 cells (t =4.00,4.71,P ＜ 0.05),but it had no obvious changes after irradiation alone.Conclusions The immune tolerance of lung cancer cells microenvironment under irradiation might contribute to the up-promotion of CD4 + CD25 + Treg cells,CTLA-4 and IL-17 although the molecular mechanisms still need to be further investigated.
