Zhang Xiao, Zhou Hua, Dong Guang-fu, Shi Yun-zhen
Department of Rheumatology, Guangdong Provincial People's Hospital, Guangzhou 510080, China.
Zhonghua Nei Ke Za Zhi. 2005 May;44(5):370-3.
To detect the effects of leflunomide to phenotype and function of dendritic cells (DCs) in systemic lupus erythematosus (SLE) patients, reveal the effective mechanism inducing remission of SLE and lay a research foundation for using 'inhibit' DCs to treat SLE in future.
The monocytes were isolated from peripheral blood of SLE patients and cultivated into DCs with cytokines such as GM-CSF and IL-4. A771726 (active metabolite) was added in with cytokines in leflunomide group, but not in control. DCs were harvested after 9 days culture. CD(80), CD(83), CD(86) and HLA-DR surface markers on DCs were detected by flow cytometry (FACS). The ability of DCs stimulating lymphocytes proliferation was detected by MTT assay. IL-10 and IFNgamma level in the supernatant of MLR were detected by ELISA and T cell subtype after MLR was detected by FACS.
The DCs treated with A771726 showed a lower percentage expression of CD(83), CD(86) and HLA-DR phenotype (CD(83): 72.70 +/- 1.77 vs. 79.36 +/- 4.80, CD(86): 63.50 +/- 14.06 vs. 83.91 +/- 9.81, HLA-DR: 80.44 +/- 12.56 vs. 90.51 +/- 8.63, all P < 0.01), a weaker ability to stimulating T lymphocytes proliferation (at DC:TC = 1:10, 0.285 +/- 0.079 vs. 0.458 +/- 0.100; at DC:TC = 1:50, 0.194 +/- 0.054 vs. 0.382 +/- 0.023, all P < 0.01) and a lower secretive level of IL-10 in the MLR supernatant [(195.0 +/- 36.9) microg/L vs. (423.6 +/- 93.2) microg/L, P < 0.01], exclude those it could still increase amount of a new T cell subtype--CD(4)(+)CD(25)(+)CTLA(4)(+) T cell (12.00% & 6.23%).
A771726 can inhibit DCs maturation, the immature DCs can inhibit T cells proliferation and refrain T cells from dividing into Th(2) subtype, and also the immature DCs can induce a sort of regulate T cell (CD(4)(+)CD(25)(+)CTLA(4)(+) T cell) production. Through that LEF may correct part over humor immune dysfunction and get a new immune balance in SLE.
检测来氟米特对系统性红斑狼疮（SLE）患者树突状细胞（DCs）表型及功能的影响，揭示其诱导SLE病情缓解的作用机制，为今后应用“抑制”DCs治疗SLE奠定研究基础。
从SLE患者外周血中分离单核细胞，用GM-CSF和IL-4等细胞因子培养成DCs。来氟米特组在细胞因子中加入A771726（活性代谢产物），对照组不加。培养9天后收获DCs。采用流式细胞术（FACS）检测DCs表面CD(80)、CD(83)、CD(86)和HLA-DR标志物。采用MTT法检测DCs刺激淋巴细胞增殖的能力。采用ELISA法检测混合淋巴细胞反应（MLR）上清液中IL-10和IFNγ水平，采用FACS检测MLR后T细胞亚群。
用A771726处理的DCs显示CD(83)、CD(86)和HLA-DR表型的表达百分比降低（CD(83)：72.70±1.77对79.36±4.80，CD(86)：63.50±14.06对83.91±9.81，HLA-DR：80.44±12.56对90.51±8.63，均P<0.01），刺激T淋巴细胞增殖的能力较弱（在DC:TC=1:10时，0.285±0.079对0.458±0.100；在DC:TC=1:50时，0.194±0.054对0.382±0.023，均P<0.01），MLR上清液中IL-10的分泌水平较低[（195.0±36.9）μg/L对（423.6±93.2）μg/L，P<0.01]，除此之外，它仍可增加一种新的T细胞亚群——CD(4)(+)CD(25)(+)CTLA(4)(+)T细胞的数量（12.00%和6.23%）。
A771726可抑制DCs成熟，未成熟的DCs可抑制T细胞增殖并阻止T细胞分化为Th(2)亚群，且未成熟的DCs可诱导产生一种调节性T细胞（CD(4)(+)CD(25)(+)CTLA(4)(+)T细胞）。由此，来氟米特可能纠正SLE部分过度的体液免疫功能紊乱并建立新的免疫平衡。
