CAJ | 학술논문

　　目的探讨系统性红斑狼狼疮（SLE）患者树突状细胞对Th1/Th2细胞亚群分化的影响。方法淋巴细胞分离液分离获得SLE患者外周血单个核细胞,免疫磁珠阳性分选CD14细胞,联合应用rhGM- CSF、rhIL-4和rhTNF-α诱导分化树突状细胞成熟。在培养的第9天，流式细胞术检测表型,细胞增殖/毒性剂盒分析树突状细胞刺激同种异体淋巴细胞增殖的能力。ELISA法检测培养上清液中IL-12和IL-10的表达。诱导的成熟树突状细胞与健康对照组淋巴细胞共培养5d后，佛波酯+离子霉素+莫能霉素刺激培养4h，进行T淋巴细胞表面抗原及胞质内细胞因子染色，流式细胞仪检测Th1、Th2的比例。结果 SLE患者诱导培养的树突状细胞其CD1a的表达降低（P＜0.05），HLA- DR、CD80、CD86的表达均明显升高（P＜0.05）。活动组与非活动组的树突状细胞HLA- DR、CD80、CD86的表达有统计学差异（P＜0.05）。SLE患者诱导培养的树突状细胞刺激同种异体淋巴细胞增殖能力增强（P＜0.05）。活动组与非活动组的树突状细胞刺激同种异体淋巴细胞也有所差异（P＜0.05）。SLE患者诱导培养的树突状细胞分泌IL-12的水平均明显降低（P＜0.05）；分泌IL-10的水平明显升高（P＜0.01），活动组与非活动组的树突状细胞分泌IL-12和IL-10的水平差异无统计学意义（P＞0.05）。活动组的树突状细胞与对照组淋巴细胞共培养后Th1的比例明显降低（P＜0.01），Th2的比例差异无统计学意义（P＞0.05）。结论 SLE患者诱导的树突状细胞表型增加，刺激同种异体淋巴细胞增殖的能力增强，分泌IL-12降低，IL-10升高。SLE患者树突状细胞功能的异常可能导致了Th1细胞分化不足，在SLE发生、发展过程中起重要作用。
　　목적탐토계통성홍반랑랑창（SLE）환자수돌상세포대Th1/Th2세포아군분화적영향。방법림파세포분리액분리획득SLE환자외주혈단개핵세포,면역자주양성분선CD14세포,연합응용rhGM- CSF、rhIL-4화rhTNF-α유도분화수돌상세포성숙。재배양적제9천，류식세포술검측표형,세포증식/독성제합분석수돌상세포자격동충이체림파세포증식적능력。ELISA법검측배양상청액중IL-12화IL-10적표체。유도적성숙수돌상세포여건강대조조림파세포공배양5d후，불파지+리자매소+막능매소자격배양4h，진행T림파세포표면항원급포질내세포인자염색，류식세포의검측Th1、Th2적비례。결과 SLE환자유도배양적수돌상세포기CD1a적표체강저（P＜0.05），HLA- DR、CD80、CD86적표체균명현승고（P＜0.05）。활동조여비활동조적수돌상세포HLA- DR、CD80、CD86적표체유통계학차이（P＜0.05）。SLE환자유도배양적수돌상세포자격동충이체림파세포증식능력증강（P＜0.05）。활동조여비활동조적수돌상세포자격동충이체림파세포야유소차이（P＜0.05）。SLE환자유도배양적수돌상세포분비IL-12적수평균명현강저（P＜0.05）；분비IL-10적수평명현승고（P＜0.01），활동조여비활동조적수돌상세포분비IL-12화IL-10적수평차이무통계학의의（P＞0.05）。활동조적수돌상세포여대조조림파세포공배양후Th1적비례명현강저（P＜0.01），Th2적비례차이무통계학의의（P＞0.05）。결론 SLE환자유도적수돌상세포표형증가，자격동충이체림파세포증식적능력증강，분비IL-12강저，IL-10승고。SLE환자수돌상세포공능적이상가능도치료Th1세포분화불족，재SLE발생、발전과정중기중요작용。
Objective To investigate the effect of dendritic cells (DC) on Th1/Th2 differentiation in patients with systemic lupus erythematosus(SLE). Methods Peripheral blood mononuclear cells (PBMC) were isolated and purified by a high gradient magnetic cell sorting system (MACS). GM- CSF, IL- 4 and TNF- αwere used to induce matured DCs, the phenotype of DCs was analyzed by immunofluorescence staining and flow cytometry9d after induction. The ability of inducing proliferation of al ogenic lymphocytes was examined by cell counting Kit- 8(CCK- 8). The production of IL- 12 and IL- 10 by the induced mature DCs were measured by ELISA. The lymphocytes in PBMC were separated by the adherent method, and were co- cultured with the matured DC from SLE patients. After 5 days, CD4+T cells were stimulated with PMA+ionomycin+monensin and the differentiation of Th1/Th2 cells was detected by flow cytometry using intracel ular fluorescent staining of IFN- γ and IL- 4. Results The expresses of CD80, CD86 and HLA- DR in DCs from SLE patients were increased(P<0.05), that of CD1a was decreased(P<0.05). There were significant differences in expression of HLA- DR, CD80, CD86 and ability of inducing al ogenic lymphocytes proliferation between active and inactive cases (P<0.05). The levels of IL- 12 were reduced and IL- 10 increased significantly in the induced mature DCs from SLE patients. There was no significant difference between active and inactive cases. In comparison to health controls, the percentage of Th1 cells in CD4+T was lower when co- cultured with the matured DC from SLE patients. Conclusion The re-sults indicate that Th1 cell differentiation is impaired in patients with SLE, which may be associated with the dysfunctions of den-dritic cells.