中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
1期
66-70
,共5页
索拉非尼%T淋巴细胞
索拉非尼%T淋巴細胞
색랍비니%T림파세포
Sorafenib%T cells
目的 阐明索拉非尼抑制正常人外周血T淋巴细胞的分子机制.方法 羧基荧光素二醋酸盐琥珀酰亚胺酯(5,6-carboxyfluorescein diacetate succinimidyl ester,CFSE)增殖试验和MTS[3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲酯基)-2-(4-磺苯基)-2H-四唑(金翁),内盐]法检测索拉非尼对T细胞的增殖和生存能力的影响;Annexin V和PI双染法检测T细胞的凋亡;FACS检测细胞周期和CD25、CD69的表达,Western blot检测细胞周期蛋白的表达;ELISA法检测IL-2的水平;采用苦基氯(picryl chloride,PCL)诱导的迟发性过敏反应(DTH)模型,检测索拉非尼对小鼠体内T细胞免疫应答的影响.结果 索拉非尼剂量依赖性的抑制T淋巴细胞的增殖,却不影响T细胞的凋亡,它可以使T细胞周期阻滞在G_0/G_1期.索拉非尼可以抑制T细胞CD25、CD69的表达,抑制IL-2的分泌,在体内索拉非尼能够抑制PCL诱导DTH模型.结论 索拉非尼可以抑制外周血T淋巴细胞的增殖和活化,临床上长期用药时可能会导致机体的免疫抑制作用.
目的 闡明索拉非尼抑製正常人外週血T淋巴細胞的分子機製.方法 羧基熒光素二醋痠鹽琥珀酰亞胺酯(5,6-carboxyfluorescein diacetate succinimidyl ester,CFSE)增殖試驗和MTS[3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲酯基)-2-(4-磺苯基)-2H-四唑(金翁),內鹽]法檢測索拉非尼對T細胞的增殖和生存能力的影響;Annexin V和PI雙染法檢測T細胞的凋亡;FACS檢測細胞週期和CD25、CD69的錶達,Western blot檢測細胞週期蛋白的錶達;ELISA法檢測IL-2的水平;採用苦基氯(picryl chloride,PCL)誘導的遲髮性過敏反應(DTH)模型,檢測索拉非尼對小鼠體內T細胞免疫應答的影響.結果 索拉非尼劑量依賴性的抑製T淋巴細胞的增殖,卻不影響T細胞的凋亡,它可以使T細胞週期阻滯在G_0/G_1期.索拉非尼可以抑製T細胞CD25、CD69的錶達,抑製IL-2的分泌,在體內索拉非尼能夠抑製PCL誘導DTH模型.結論 索拉非尼可以抑製外週血T淋巴細胞的增殖和活化,臨床上長期用藥時可能會導緻機體的免疫抑製作用.
목적 천명색랍비니억제정상인외주혈T림파세포적분자궤제.방법 최기형광소이작산염호박선아알지(5,6-carboxyfluorescein diacetate succinimidyl ester,CFSE)증식시험화MTS[3-(4,5-이갑기새서-2-기)-5-(3-최갑지기)-2-(4-광분기)-2H-사서(금옹),내염]법검측색랍비니대T세포적증식화생존능력적영향;Annexin V화PI쌍염법검측T세포적조망;FACS검측세포주기화CD25、CD69적표체,Western blot검측세포주기단백적표체;ELISA법검측IL-2적수평;채용고기록(picryl chloride,PCL)유도적지발성과민반응(DTH)모형,검측색랍비니대소서체내T세포면역응답적영향.결과 색랍비니제량의뢰성적억제T림파세포적증식,각불영향T세포적조망,타가이사T세포주기조체재G_0/G_1기.색랍비니가이억제T세포CD25、CD69적표체,억제IL-2적분비,재체내색랍비니능구억제PCL유도DTH모형.결론 색랍비니가이억제외주혈T림파세포적증식화활화,림상상장기용약시가능회도치궤체적면역억제작용.
Objective To elucidate the molecular mechanisms of sorafenib inhibitting human pe-ripheral blood T cells. Methods CFSE(5, 6-carboxyfluorescein diacetate succinimidyl ester) proliferation assay and MTS [3-(4, 5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl) -2-( 4-sulfophenyl) -2H-etrazoli-um, inner salt] assay were used to examine the proliferation and the viability of T cells; Annexin V-FITC and PI staining was used to detect apoptosis; Flow cytometry was used to detect the expression of CD25, CD69; Western blot was used to detect the expression of cell cycle proteins; ELISA was used to detect the level of IL-2; Picryl chloride-induced delayed-type hypersensitivity model to be used to for the evaluation of in vivo immunocompetency. Results Sorafenib inhibited proliferation of human peripheral blood T cells in-duced by phytohemagglutinin(PHA) in a dose-dependent manner without inducing their apeptosis. Sorafenib caused human blood T cells arrest in the G_0/G_1 phase of the ceLl cycle. Sorafenib decreased CD25 and CD69 expressions and IL-2 production in human T cells. Sorafenib inhibited picryl chloride-induced delayed-type hypersensitivity in mice. Conclusion Sorafenib could inhibit proliferation and activation of peripheral blood T cells. These finding indicated that long term administration of sorafenib might lead to immunosuppressive effects.
