他克莫司体外诱导狼疮性肾炎患者耐受性树突状细胞形成  

In Vitro Effect of Tacrolimus on Inducing Tolerogenic Dendritic Cells in Lupus Nephritis Patients

作　　者：刘冠贤[1] 石咏军[1] 

机构地区：[1]广东省惠州市中心医院肾内科,惠州516001

出　　处：《中国中西医结合肾病杂志》2006年第7期382-385,共4页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：广东省自然科学基金资助项目(No.04300905)

摘　　要：目的:观察他克莫司(tacrolimus)处理前后狼疮性肾炎(LN)患者外周血树突状细胞(dendriticcells,DCs)表面标志及功能的改变。方法:分离LN患者外周血单个核细胞,用GM-CSF、IL-4等细胞因子诱导DCs成熟,他克莫司组在加入上述细胞因子前先加入他克莫司(5μg/ml)培养。培养第9d收集DCs细胞,流式细胞仪检测CD80、CD86和HLA-DR的表达。MTT法检测DCs刺激淋巴细胞增殖能力,ELISA法检测混合淋巴细胞反应培养上清IL-10和IFN-γ水平。结果:他克莫司处理后的DCs表达CD80、CD86和HLA-DR百分数较对照组均明显降低(52.70±1.77vs78.36±4.80,63.50±14.06vs83.91±9.81,70.41±12.51vs90.51±8.63),P均<0.01;他克莫司处理后的DCs与T细胞混合培养,其刺激T细胞增殖相应的OD值明显降低(DC∶TC=1∶10时,0.294±0.094vs0.582±0.123;DC∶TC=1∶50时,0.325±0.099vs0.458±0.080),P均<0.01。其混合培养的上清液中IL-10水平较无他克莫司处理的DCs与T细胞的混合培养上清液明显降低[(195.0±36.9)pg/mlvs(423.6±93.2)pg/ml,P<0.01],而IFN-γ两者间无统计学意义[(88.2±11.6)pg/mlvs(86.9±12.7)pg/ml,P>0.05]。结论:他克莫司在体外可抑制LN患者外周血DCs的成熟,且他克莫司处理后的DCs能明显抑制T细胞增殖及T细胞向Th2细胞转化。Objective： To determine the effects of tacrolimus on the phenotype and immune function of peripheral blood dendritic cells （DCs） in lupus nephritis（LN） patients. Methods： The mononuclear cells were isolated from peripheral blood in LN patients and developed into DCs with cytokines such as GM-CSF and IL-4. In tacrolimus group, tacrolimus （5μg/ml）was added before the above cytokines was used. DCs were harvested after 9 days culture. CD80, CD86 and HLA-DR surface markers on DCs were detected by flow cytometry. MTT assay detected mixed lymphocyte reaction （MLR）. IL- 10 and IFN-γ in the supermatant of MLR were detected by ELISA. Results： The percentage of CD80 ,CD86 and HLA-DR expression on the DCs treated with tacrolimus were lower than them on the DCs without tacrolimus-treated（ （52.70±1.77 vs 78.36±4.80, 63.50± 14.06 vs 83.91±9.81,70.41±12.51 vs 90.51±8.63）,all P〈0.01. The OD value related with T lymphocytes proliferation were lower in the MLR with the with treated with tacrolimus than in the MLR with the without tacrolimus-treated（at DC：TC= 1：10, 0.29±0.094 vs 0.582±0. 123：at DC：TC= 1：50,0.325＋0.099 vs 0. 458±0. 080）,both P value〈0.01. IL-10 level in the MLR supermatant of the group treated with tacrolimus was lower than in the control group（ 195.0 ±36.9 pg/ml vs 423.6±93.2 pg/ml, P〈 0.01）., but IFN-γ level in both MLR supernatant have no significant difference（88.2±11.6 pg/ml vs 86.9±12.7 pg/ml, P 〉0.05）. Conclusion： Tacrolimus can inhibits can in maturation, and then the immature then can also inhibit T cells proliferation and refrain T cells from differentiating into Th2 subtype.

关 键 词：树突状细胞 T淋巴细胞 他克莫司 狼疮性肾炎 

分 类 号：R593.242[医药卫生—内科学]