中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2011年
12期
834-838
,共5页
宗明%张慧%孙立山%金中淦%范列英
宗明%張慧%孫立山%金中淦%範列英
종명%장혜%손립산%금중감%범렬영
波形蛋白%树突细胞%关节炎,类风湿%瓜氨酸化
波形蛋白%樹突細胞%關節炎,類風濕%瓜氨痠化
파형단백%수돌세포%관절염,류풍습%과안산화
Vimentin%Dendritic cells%Arthritis,rheumatoid%Citruilination
目的 探讨波形蛋白瓜氨酸化前后在体外对类风湿关节炎(RA)患者外周血来源树突状细胞( DCs)的细胞形态、表型和功能的影响.方法 分离外周血单个核细胞(PBMCs),粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素(IL)-4体外培养制备未成熟DCs(imDCs),分别用脂多糖、瓜氨酸化波形蛋白(cVim)和波形蛋白刺激培养的imDCs,磷酸盐缓冲液(PBS)作为阴性对照,流式细胞仪检测DCs表面标志CD14、CD80、CD83、CD86、主要组织相容性复合体(MHC)Ⅰ、MHCⅡ表达变化.将实验组获得的DCs和同种异体T细胞进行混合淋巴细胞反应,通过单溶液细胞增殖分析(MTS)法检测T细胞的增殖情况.采用t检验.结果 以阴性对照组imDCs的各个免疫表型阳性率作为1,脂多糖能显著诱导RAimDCs表面MHCⅡ、CD80、CD83、CD86表达(1.07±0.14,1.25±0.13,1.90±1.08,2.44±0.65,P均＜0.05)；cVim诱导MHCⅡ(1.18±0.09)、CD83( 1.97±0.99)表达水平上调(P＜0.05,P＜0.01)；波形蛋白对MHCⅡ及CD83、CD86表达无影响(0.950.11,0.90±0.29,1.04±0.06),仅抑制CD80表达(0.82±0.18,P＜0.01).与脂多糖组相比,cVim组的MHCⅡ表达水平显著增高(P＜0.05),而CD80、CD86(0.83±0.36、1.32±0.15)的表达水平则低于脂多糖组(P＜0.01,P＜0.05)；cVim组MHCⅡ和CD83的表达水平显著高于波形蛋白组(P均＜0.05).混合淋巴细胞反应显示经脂多糖和cVim诱生的DCs对同种异体T细胞具有明显刺激增殖作用,且随着DCs细胞浓度的增加而作用增强；波形蛋白诱生的DCs则无刺激增殖作用.结论cVim可诱导imDCs成熟,并且能够提高细胞表面共刺激分子的表达.
目的 探討波形蛋白瓜氨痠化前後在體外對類風濕關節炎(RA)患者外週血來源樹突狀細胞( DCs)的細胞形態、錶型和功能的影響.方法 分離外週血單箇覈細胞(PBMCs),粒細胞-巨噬細胞集落刺激因子(GM-CSF)和白細胞介素(IL)-4體外培養製備未成熟DCs(imDCs),分彆用脂多糖、瓜氨痠化波形蛋白(cVim)和波形蛋白刺激培養的imDCs,燐痠鹽緩遲液(PBS)作為陰性對照,流式細胞儀檢測DCs錶麵標誌CD14、CD80、CD83、CD86、主要組織相容性複閤體(MHC)Ⅰ、MHCⅡ錶達變化.將實驗組穫得的DCs和同種異體T細胞進行混閤淋巴細胞反應,通過單溶液細胞增殖分析(MTS)法檢測T細胞的增殖情況.採用t檢驗.結果 以陰性對照組imDCs的各箇免疫錶型暘性率作為1,脂多糖能顯著誘導RAimDCs錶麵MHCⅡ、CD80、CD83、CD86錶達(1.07±0.14,1.25±0.13,1.90±1.08,2.44±0.65,P均＜0.05)；cVim誘導MHCⅡ(1.18±0.09)、CD83( 1.97±0.99)錶達水平上調(P＜0.05,P＜0.01)；波形蛋白對MHCⅡ及CD83、CD86錶達無影響(0.950.11,0.90±0.29,1.04±0.06),僅抑製CD80錶達(0.82±0.18,P＜0.01).與脂多糖組相比,cVim組的MHCⅡ錶達水平顯著增高(P＜0.05),而CD80、CD86(0.83±0.36、1.32±0.15)的錶達水平則低于脂多糖組(P＜0.01,P＜0.05)；cVim組MHCⅡ和CD83的錶達水平顯著高于波形蛋白組(P均＜0.05).混閤淋巴細胞反應顯示經脂多糖和cVim誘生的DCs對同種異體T細胞具有明顯刺激增殖作用,且隨著DCs細胞濃度的增加而作用增彊；波形蛋白誘生的DCs則無刺激增殖作用.結論cVim可誘導imDCs成熟,併且能夠提高細胞錶麵共刺激分子的錶達.
목적 탐토파형단백과안산화전후재체외대류풍습관절염(RA)환자외주혈래원수돌상세포( DCs)적세포형태、표형화공능적영향.방법 분리외주혈단개핵세포(PBMCs),립세포-거서세포집락자격인자(GM-CSF)화백세포개소(IL)-4체외배양제비미성숙DCs(imDCs),분별용지다당、과안산화파형단백(cVim)화파형단백자격배양적imDCs,린산염완충액(PBS)작위음성대조,류식세포의검측DCs표면표지CD14、CD80、CD83、CD86、주요조직상용성복합체(MHC)Ⅰ、MHCⅡ표체변화.장실험조획득적DCs화동충이체T세포진행혼합림파세포반응,통과단용액세포증식분석(MTS)법검측T세포적증식정황.채용t검험.결과 이음성대조조imDCs적각개면역표형양성솔작위1,지다당능현저유도RAimDCs표면MHCⅡ、CD80、CD83、CD86표체(1.07±0.14,1.25±0.13,1.90±1.08,2.44±0.65,P균＜0.05)；cVim유도MHCⅡ(1.18±0.09)、CD83( 1.97±0.99)표체수평상조(P＜0.05,P＜0.01)；파형단백대MHCⅡ급CD83、CD86표체무영향(0.950.11,0.90±0.29,1.04±0.06),부억제CD80표체(0.82±0.18,P＜0.01).여지다당조상비,cVim조적MHCⅡ표체수평현저증고(P＜0.05),이CD80、CD86(0.83±0.36、1.32±0.15)적표체수평칙저우지다당조(P＜0.01,P＜0.05)；cVim조MHCⅡ화CD83적표체수평현저고우파형단백조(P균＜0.05).혼합림파세포반응현시경지다당화cVim유생적DCs대동충이체T세포구유명현자격증식작용,차수착DCs세포농도적증가이작용증강；파형단백유생적DCs칙무자격증식작용.결론cVim가유도imDCs성숙,병차능구제고세포표면공자격분자적표체.
Objective To study the effects of citrullinated vimentin (cVim) on the maturation and immunologic function of dendritic cells (DCs) from rheumatoid arthritis (RA) peripheral blood.Methods In the present study,mononuclear cells were isolated from the peripheral blood of patients with RA and cultivated in media containing GM-CSF and IL-4 to generate immature DCs (imDCs).The imDCs generated were stimulated with citrullinated vimentin and vimentin.LPS was used as the positive control and PBS was used as the negative control.The expression of surface molecules on the DCs,such as CD14,CD80,CD83,CD86,MHC Ⅰ and MHC Ⅱ were analyzed with FACS.The capability of the stimulatory activity of the DCs on allogeneic T cells in mixed reaction was tested by MTS.t-test was used for statistical analysis.Results Compared to untreated DCs,DCs treated with LPS increased the expression levels of MHC Ⅱ,CD80,CD83 and CD86 (1.07±0.14,1.25±0.13,1.90±1.08,2.44±0.65,P＜0.05),while cVim increased the expression levels of MHC Ⅱ ( 1.18±0.09,P＜0.05) and CD83 ( 1.97±0.99,P＜0.01 ),and Vim decreased the expression levels of CD80 (0.82±0.18,P＜0.01 ).It was demonstrated that the expression levels of MHC Ⅱ on DCs pulsed with cVim were significantly higher than that of the DCs with LPS,but the expression levels of CD80 and CD86 were not significantly different.The expression levels of MHC Ⅱ and CD83 on DCs pulsed with cVim were significantly higher than that of the DCs with Vim.The mixed lymphocyte reaction showed that the DCs induced by LPS and cVim trigerred the proli-feration of allogenic T cells obviously.Conclusion This result suggests that cVim could promote the phenotypic maturation of DCs and increase the expression of costimulatory molecules.
