CAJ | 학술논문

目的通过体外实验观察白细胞介素(IL)-21及地塞米松对系统性红斑狼疮(SLE)患者滤泡辅助性T细胞(Tfh)表达的调控作用并初步探讨其可能的机制.方法收集11例SLE患者外周血,将分离出的单个核细胞(PBMCs)按106个/孔置入96孔培养板,分对照组、重组人白细胞介素(rIL)-21组及rIL-21+地塞米松组(rIL-21浓度为200 ng/μl、地塞米松浓度为1×10-6 mol/L)体外分别培养24、72 h；另分离20例SLE患者PBMCs,分对照组及地塞米松组(地塞米松浓度分别为0.5×10-7、1×10-6mol/L)体外培养24 h.收集细胞以CD4、PD1、CXCR5流式检测Tfh的百分比,酶联免疫吸附试验(ELISA)法检测上清中抗核抗体的表达.配对t检验行2组间差异比较.结果与对照组[24 h(4.3±1.2)％,72 h(5.6±4.0)％]相比,体外培养24h(5.9±2.4)％及72 h后(8.0±5.6)％rIL-21组Tfh细胞的百分比显著升高,差异有统计学意义(t=3.48和3.05,P=0.007和0.012);与rIL-21组相比,培养24 h(4.2±1.3)％及72 h(6.2±4.4)％后rIL-21+地塞米松组Tfh细胞的百分比有所下降,但仍高于对照组,差异有统计学意义(t=4.70和2.78,P=0.001和0.019)；地塞米松显著降低Tfh细胞的百分比,并具有浓度依赖性[对照组(4.0±1.2)％,低浓度地塞米松组(3.3±1.2)％,高浓度地塞米松(2.9±1.0)％,低浓度地塞米松组与对照组相比差异有统计学意义(t=3.37,P=0.003)；高浓度地塞米松组与低浓度地塞米松组相比差异有统计学意义(t=2.26,P=0.036).各组上清中抗核抗体差异无统计学意义(P＞0.05).结论 IL-21能够增加SLE患者Tfh细胞比例,而地塞米松对Tfh细胞的产生具有抑制作用,且可对抗IL-21诱导的Tfh细胞增殖.
목적 통과체외실험관찰백세포개소(IL)-21급지새미송대계통성홍반랑창(SLE)환자려포보조성T세포(Tfh)표체적조공작용병초보탐토기가능적궤제.방법 수집11례SLE환자외주혈,장분리출적단개핵세포(PBMCs)안106개/공치입96공배양판,분대조조、중조인백세포개소(rIL)-21조급rIL-21+지새미송조(rIL-21농도위200 ng/μl、지새미송농도위1×10-6 mol/L)체외분별배양24、72 h；령분리20례SLE환자PBMCs,분대조조급지새미송조(지새미송농도분별위0.5×10-7、1×10-6mol/L)체외배양24 h.수집세포이CD4、PD1、CXCR5류식검측Tfh적백분비,매련면역흡부시험(ELISA)법검측상청중항핵항체적표체.배대t검험행2조간차이비교.결과 여대조조[24 h(4.3±1.2)％,72 h(5.6±4.0)％]상비,체외배양24h(5.9±2.4)％급72 h후(8.0±5.6)％rIL-21조Tfh세포적백분비현저승고,차이유통계학의의(t=3.48화3.05,P=0.007화0.012);여rIL-21조상비,배양24 h(4.2±1.3)％급72 h(6.2±4.4)％후rIL-21+지새미송조Tfh세포적백분비유소하강,단잉고우대조조,차이유통계학의의(t=4.70화2.78,P=0.001화0.019)；지새미송현저강저Tfh세포적백분비,병구유농도의뢰성[대조조(4.0±1.2)％,저농도지새미송조(3.3±1.2)％,고농도지새미송(2.9±1.0)％,저농도지새미송조여대조조상비차이유통계학의의(t=3.37,P=0.003)；고농도지새미송조여저농도지새미송조상비차이유통계학의의(t=2.26,P=0.036).각조상청중항핵항체차이무통계학의의(P＞0.05).결론 IL-21능구증가SLE환자Tfh세포비례,이지새미송대Tfh세포적산생구유억제작용,차가대항IL-21유도적Tfh세포증식.
Objective To investigate the effect of interleukin(IL)-21 and dexamethasone(Dex)treatment on the percentage of follicular helper T cells(Tfh)in patients with systemic lupus erythematosus (SLE).Methods Peripheral blood mononuclear cells(PBMCs)obtained from 11 SLE patients were cultured in 96 well plates at 106/well with medium as the control,and with rIL-21(200 ng/μl)or rIL-21(200 ng/μl)+Dex(1×10-6 mol/L)for 24 and 72 hours as the study groups.Another 20 samples were collected and incubated with different concentrations of Dex(0,5×10-7 mol/L,1×10-6 mol/L)for 24 hours.After labeling with CD4,PD1 and CXCR5 monoclonal antibodies,the proportions of Tfh cells were assessed by flow cytometry.ELISA was used to detect supernatant antinuclear antibody(ANA)levels.Differences between groups were analyzed by paired t test.Results The percentages of Tfb cells were elevated when incubated with rIL-21 for 24 h and 72 h[24 h(4.3±1.2)％,(5.9±2.4)％;72 h(5.6±4.0)％,(8.0±5.6)％;t=3.48 and 3.05 respectively,P=0.007 and 0.012].Compared with those in the rIL-21 group,the percentages of Tfh cells in the rIL-21+Dex group were decreased(t=4.70 and 2.78,P=0.001 and 0.019).Tfh cells were decreased ina dose-dependent manner when treated with Dex[24 h(4.2±1.3)％;72 h(6.2±4.4)％;t=3.37 and 2.26,P=0.003 and 0.036].There was no difference of supe(rn)atant ANA levels among those groups.Conclusion IL-21 can promote the proliferation of Tfh cells,while Dex inhibits Tfh cell growth and the inhibition effect is dose dependent.Tfh cells may represent a new target for the treatment of SLE.