医学信息
醫學信息
의학신식
MEDICAL INFORMATION
2013年
27期
230-232
,共3页
周红娟%蔡龙%郑红霞%施华萍%商学钗%马纪林%张风卫
週紅娟%蔡龍%鄭紅霞%施華萍%商學釵%馬紀林%張風衛
주홍연%채룡%정홍하%시화평%상학차%마기림%장풍위
系统性红斑狼疮%树突状细胞%调节性%T细胞
繫統性紅斑狼瘡%樹突狀細胞%調節性%T細胞
계통성홍반랑창%수돌상세포%조절성%T세포
Systemic lupus erythematosus%Dendritic cells%Regulatory%T cells
目的：探讨系统性红斑狼疮（SLE）患者树突状细胞（DC）对CD4+CD25+调节性 T细胞分化的影响以及两者之间的关系。方法淋巴细胞分离液分离获得SLE患者外周血单个核细胞（PBMC），免疫磁珠阳性分选CD14+细胞，细胞因子诱导获得 iDCs和 mDCs。在培养的第9d，流式细胞术检测表型，细胞增殖/毒性(CCK-8)试剂盒分析iDCs和mDCs分别刺激同种异体淋巴细胞活化增殖的能力。分选健康对照组 CD4+CD25-T细胞，分别与 iDCs和 mDCs共培养，5d后流式检测CD4+CD25+Foxp3+细胞所占的比例。酶联免疫吸附试验（ELISA）方法检测混合培养上清中 IL-10和 TGF-β的浓度。结果表型分析发现，各组 mDCs表面标志物的表达均高于iDC组；SLE活动组mDCs表面标志物 CD80、CD86的表达较健康对照组 mDCs有明显上升；iDCs表面标志物的表达较健康对照组iDCs也有明显上升。分别与健康对照mDCs和iDCs比较，SLE活动组刺激同种异体淋巴细胞活化增殖能力均增强；SLE活动组 mDCs明显强于 iDCs。健康对照组CD4+CD25-T细胞与各组DCs共培养后，SLE活动组 iDCs 的 CD4+CD25+Foxp3+比例较健康对照组 iDCs下降，mDC组的比例较健康对照组 mDC组上升。 SLE活动组iDCs和mDCs分泌的IL-10和TGF-β均低于健康对照组，SLE非活动组 iDC组分泌的 IL-10和 TGF-β也均低于健康对照组 iDC组。 SLE活动组iDC组分泌的IL-10、TGF-β和mDC组分泌的TGF-β均低于非活动组对应组。分泌的细胞因子进行组内之间比较发现，健康对照组分泌的 IL-10和TGF-β、SLE活动组和非活动组分泌的 TGF-β，其mDC组均低于 iDC组。结论 SLE活动组 mDCs表面标志物 CD80、CD86的表达升高，其 iDCs和 mDCs刺激同种异体淋巴细胞活化增殖能力增强。健康对照组CD4+CD25-T细胞与 SLE活动组 iDCs 共培养后，CD4+CD25+Foxp3+比例下降，mDC组的比例上升。SLE活动组iDCs和mDCs分泌的IL-10和TGF-β均降低。此结果表明 DCs与 Treg的相互影响在 DCs诱导免疫耐受机制中扮演重要角色，对于 SLE 的发病和治疗机制可能会有突破性进展。
目的：探討繫統性紅斑狼瘡（SLE）患者樹突狀細胞（DC）對CD4+CD25+調節性 T細胞分化的影響以及兩者之間的關繫。方法淋巴細胞分離液分離穫得SLE患者外週血單箇覈細胞（PBMC），免疫磁珠暘性分選CD14+細胞，細胞因子誘導穫得 iDCs和 mDCs。在培養的第9d，流式細胞術檢測錶型，細胞增殖/毒性(CCK-8)試劑盒分析iDCs和mDCs分彆刺激同種異體淋巴細胞活化增殖的能力。分選健康對照組 CD4+CD25-T細胞，分彆與 iDCs和 mDCs共培養，5d後流式檢測CD4+CD25+Foxp3+細胞所佔的比例。酶聯免疫吸附試驗（ELISA）方法檢測混閤培養上清中 IL-10和 TGF-β的濃度。結果錶型分析髮現，各組 mDCs錶麵標誌物的錶達均高于iDC組；SLE活動組mDCs錶麵標誌物 CD80、CD86的錶達較健康對照組 mDCs有明顯上升；iDCs錶麵標誌物的錶達較健康對照組iDCs也有明顯上升。分彆與健康對照mDCs和iDCs比較，SLE活動組刺激同種異體淋巴細胞活化增殖能力均增彊；SLE活動組 mDCs明顯彊于 iDCs。健康對照組CD4+CD25-T細胞與各組DCs共培養後，SLE活動組 iDCs 的 CD4+CD25+Foxp3+比例較健康對照組 iDCs下降，mDC組的比例較健康對照組 mDC組上升。 SLE活動組iDCs和mDCs分泌的IL-10和TGF-β均低于健康對照組，SLE非活動組 iDC組分泌的 IL-10和 TGF-β也均低于健康對照組 iDC組。 SLE活動組iDC組分泌的IL-10、TGF-β和mDC組分泌的TGF-β均低于非活動組對應組。分泌的細胞因子進行組內之間比較髮現，健康對照組分泌的 IL-10和TGF-β、SLE活動組和非活動組分泌的 TGF-β，其mDC組均低于 iDC組。結論 SLE活動組 mDCs錶麵標誌物 CD80、CD86的錶達升高，其 iDCs和 mDCs刺激同種異體淋巴細胞活化增殖能力增彊。健康對照組CD4+CD25-T細胞與 SLE活動組 iDCs 共培養後，CD4+CD25+Foxp3+比例下降，mDC組的比例上升。SLE活動組iDCs和mDCs分泌的IL-10和TGF-β均降低。此結果錶明 DCs與 Treg的相互影響在 DCs誘導免疫耐受機製中扮縯重要角色，對于 SLE 的髮病和治療機製可能會有突破性進展。
목적：탐토계통성홍반랑창（SLE）환자수돌상세포（DC）대CD4+CD25+조절성 T세포분화적영향이급량자지간적관계。방법림파세포분리액분리획득SLE환자외주혈단개핵세포（PBMC），면역자주양성분선CD14+세포，세포인자유도획득 iDCs화 mDCs。재배양적제9d，류식세포술검측표형，세포증식/독성(CCK-8)시제합분석iDCs화mDCs분별자격동충이체림파세포활화증식적능력。분선건강대조조 CD4+CD25-T세포，분별여 iDCs화 mDCs공배양，5d후류식검측CD4+CD25+Foxp3+세포소점적비례。매련면역흡부시험（ELISA）방법검측혼합배양상청중 IL-10화 TGF-β적농도。결과표형분석발현，각조 mDCs표면표지물적표체균고우iDC조；SLE활동조mDCs표면표지물 CD80、CD86적표체교건강대조조 mDCs유명현상승；iDCs표면표지물적표체교건강대조조iDCs야유명현상승。분별여건강대조mDCs화iDCs비교，SLE활동조자격동충이체림파세포활화증식능력균증강；SLE활동조 mDCs명현강우 iDCs。건강대조조CD4+CD25-T세포여각조DCs공배양후，SLE활동조 iDCs 적 CD4+CD25+Foxp3+비례교건강대조조 iDCs하강，mDC조적비례교건강대조조 mDC조상승。 SLE활동조iDCs화mDCs분비적IL-10화TGF-β균저우건강대조조，SLE비활동조 iDC조분비적 IL-10화 TGF-β야균저우건강대조조 iDC조。 SLE활동조iDC조분비적IL-10、TGF-β화mDC조분비적TGF-β균저우비활동조대응조。분비적세포인자진행조내지간비교발현，건강대조조분비적 IL-10화TGF-β、SLE활동조화비활동조분비적 TGF-β，기mDC조균저우 iDC조。결론 SLE활동조 mDCs표면표지물 CD80、CD86적표체승고，기 iDCs화 mDCs자격동충이체림파세포활화증식능력증강。건강대조조CD4+CD25-T세포여 SLE활동조 iDCs 공배양후，CD4+CD25+Foxp3+비례하강，mDC조적비례상승。SLE활동조iDCs화mDCs분비적IL-10화TGF-β균강저。차결과표명 DCs여 Treg적상호영향재 DCs유도면역내수궤제중분연중요각색，대우 SLE 적발병화치료궤제가능회유돌파성진전。
Objective to investigate the functions of dendritic cells ( DC) and its influences on the CD4 +CD25+Treg dif erentiation in patients with systemic lupus erythematosus. Methods Peripheral blood mononuclear cells (PBMC) were isolated and purified by a high gradient magnetic cellsorting system (MACS),then matured DCs (mDCs) and immatured DCs(iDCs) were induced by cytokines. After 9 days, the phenotype of DCs was analyzed by the immunofluorescence staining and flow cytometry. The ability of inducing proliferation of al ogenic lymphocytes was examined by cellcounting Kit-8 (CCK-8). The CD4+CD25-cells of normal group NG were cultured with iDCs and mDCs respectively, and then the ratio of CD4+CD25+Foxp3+ cells was analyzed by flow cytometry. The IL-10 and TGF-β expression levels of mixed culture supernatant were measured by ELISA. Results The expresses of the surface markers of mDCs were exceed to the iDCs. The expresses of CD80, CD86 of mDCs cultured from SLE-active patients were higher than the mDCs of NG, similarly the iDCs. Compare to mDCs and iDCs of NG, the abilities of inducing proliferation of al ogenic lymphocytes were strengthened, the mDCs were stronger to the iDCs. The SLE iDCs were cultured with the CD4+CD25-cells of NG;the ratio of the CD4+CD25+Foxp3+was decreased. But the mDCs group went up. The levels of IL-10 and TGF-β were decreased in SLE-active group, and that of the SLE-inactive group were also lower than NG group. Conclusion The expresses of CD80, CD86 of mDCs cultured from SLE-active patients were increased, and the abilities of inducing proliferation of al ogenic lymphocytes were strengthened. The SLE iDCs were cultured with The CD4+CD25-cells of normal group NG, the ratio of the CD4+CD25+Foxp3+was decreased. But the mDCs group went up. The levels of IL-10 and TGF-β were decreased in SLE-active group. The results show that the interaction between DCs and Treg may play an important role in the mechanism of immune tolerance induced in SLE, which may have a breakthrough in the pathogenesis and treatment of SLE.
