CAJ | 학술논문

目的：观察白细胞介素-37(IL-37)在健康人外周血 CD4+CD25+调节性 T细胞(Treg)中的表达情况。方法：采用免疫磁性分离法分离人外周血 CD4+CD25+Treg,流式细胞技术分析其分离纯度,观察刺激剂组(每1×105个 CD4+CD25+Treg细胞加入20μl Dynabeads? Human Treg Expander)、非刺激剂组、空白对照组(单纯10%FCS-RPMI 1640培养)CD4+CD25+Treg细胞增殖活性及其差异。分别采用 Western blot和免疫荧光共聚焦技术检测 IL-37的表达水平与定位改变。结果：MACS分离人外周血 CD4+CD25+Treg 细胞纯度为93%,台盼蓝染色细胞活性为98%。与非刺激剂组和空白对照组相比,刺激剂组 CD4+CD25+Treg 增殖活性随时间延长而逐渐增加；非刺激剂组与空白对照组其增殖活性差异无显著性。Western blot 结果显示,正常CD4+CD25+Treg细胞表达 IL-37,在刺激剂作用下随作用时间延长 IL-37表达量增加。免疫荧光显微镜和免疫荧光共聚焦图像显示,刺激72 h后 IL-37在 CD4+CD25+Treg细胞内明显表达,并且在细胞质中表达丰富,近胞膜处表达密度较高。结论：IL-37在健康人外周血 CD4+CD25+Treg 细胞中表达,当受到有效刺激后 IL-37表达明显上升。
목적：관찰백세포개소-37(IL-37)재건강인외주혈 CD4+CD25+조절성 T세포(Treg)중적표체정황。방법：채용면역자성분리법분리인외주혈 CD4+CD25+Treg,류식세포기술분석기분리순도,관찰자격제조(매1×105개 CD4+CD25+Treg세포가입20μl Dynabeads? Human Treg Expander)、비자격제조、공백대조조(단순10%FCS-RPMI 1640배양)CD4+CD25+Treg세포증식활성급기차이。분별채용 Western blot화면역형광공취초기술검측 IL-37적표체수평여정위개변。결과：MACS분리인외주혈 CD4+CD25+Treg 세포순도위93%,태반람염색세포활성위98%。여비자격제조화공백대조조상비,자격제조 CD4+CD25+Treg 증식활성수시간연장이축점증가；비자격제조여공백대조조기증식활성차이무현저성。Western blot 결과현시,정상CD4+CD25+Treg세포표체 IL-37,재자격제작용하수작용시간연장 IL-37표체량증가。면역형광현미경화면역형광공취초도상현시,자격72 h후 IL-37재 CD4+CD25+Treg세포내명현표체,병차재세포질중표체봉부,근포막처표체밀도교고。결론：IL-37재건강인외주혈 CD4+CD25+Treg 세포중표체,당수도유효자격후 IL-37표체명현상승。
Objective:To investigate the expression of interleukin-37 (IL-37)in healthy human peripheral blood CD4+CD25+ regulatory T cells (Tregs).Methods:Magnetic Activated Cell Sorting (MACS)kit was used to seg-regate CD4+CD25+Tregs in human peripheral blood,and the purity of CD4+CD25+Tregs was determined by flow cytometry. Proliferation activity of CD4+CD25+Tregs was measured in expander group (1×105 CD4+CD25+Treg were cultured with 20μl Dynabeads? Human Treg expander),non-expander group,and blank control group(cells were cultured with 10%FCS-RPMI 1640).Meanwhile,the expression of IL-37 in CD4+CD25+Tregs was analyzed by laser scanning confocal microscope and Western blot,respectively. Results:The purity of CD4+CD25+Tregs was over 93% after being isolated twice by magnetic beads,and the activity of CD4+CD25+Tregs exceeded 98%. In comparison with non-expander and blank control groups,proliferation activity in expander group was signifi-cantly increased along with the prolongation of stimulation. However,the non-expander group and blank control goup showed no significant proliferation. With laser scanning confocal microscope and Western blot,it was identi-fied the expression of IL-37 in human CD4+CD25+ Tregs. Under the stimulation by expander,the expression of IL-37 in CD4+CD25+Tregs was markedly enhanced. In addition,expression of IL-37 was obvious in the cyto-plasm, and its density was found to be high near the cell membrane under laser scanning confocal microscopy.Conclusions:In healthy human peripheral blood,IL-37 can positively express in CD4+CD25+Tregs. With the stimulation by expander,the expression of IL-37 might further be enhanced in CD4+CD25+Tregs.