CAJ | 학술논문

目的观察重症肌无力(MG)患者外周血中负性协同刺激分子programmed death-1( PD-1)的表达情况，并探讨其与MG发病的关系。方法采用免疫荧光标记、流式细胞仪检测45例MG患者和33名健康对照者外周血单个核细胞中PD-1及其配体PD-L1的表达，用ELISA法检测各组血浆中可溶性PD-1的水平。结果 (1)MG患者表达PD-1的CD4+T淋巴细胞比例增加，CD14+PD-L1+的单核细胞比例增加，但在不同性别及眼肌型与全身型间差异无统计学意义；在胸腺异常MG患者中CD4+PD-1+T细胞增加，CD14+ PD-L1+的单核细胞比例减少；早发型MG患者（年龄＜40岁）CD4+PD-1+T淋巴细胞比例明显低于晚发型（年龄≥40岁）。(2)MG患者血浆中sPD-1浓度为(6.92 +0.72) ng/ml，明显高于健康对照组的(3.28±0.42) ng/ml，但在性别、MG眼肌型与全身型不同类型间和有无胸腺异常各组间差异无统计学意义，且sPD-1与发病年龄呈负相关(r=-0.526，P=0.000)。结论 PD-1及PD-L1途径参与了MG的发病，异常升高的sPD-1可能干扰了正常的细胞膜上PD-1与PD-L1的结合，从而促使疾病进展。
목적 관찰중증기무력(MG)환자외주혈중부성협동자격분자programmed death-1( PD-1)적표체정황，병탐토기여MG발병적관계。방법 채용면역형광표기、류식세포의검측45례MG환자화33명건강대조자외주혈단개핵세포중PD-1급기배체PD-L1적표체，용ELISA법검측각조혈장중가용성PD-1적수평。결과 (1)MG환자표체PD-1적CD4+T림파세포비례증가，CD14+PD-L1+적단핵세포비례증가，단재불동성별급안기형여전신형간차이무통계학의의；재흉선이상MG환자중CD4+PD-1+T세포증가，CD14+ PD-L1+적단핵세포비례감소；조발형MG환자（년령＜40세）CD4+PD-1+T림파세포비례명현저우만발형（년령≥40세）。(2)MG환자혈장중sPD-1농도위(6.92 +0.72) ng/ml，명현고우건강대조조적(3.28±0.42) ng/ml，단재성별、MG안기형여전신형불동류형간화유무흉선이상각조간차이무통계학의의，차sPD-1여발병년령정부상관(r=-0.526，P=0.000)。결론 PD-1급PD-L1도경삼여료MG적발병，이상승고적sPD-1가능간우료정상적세포막상PD-1여PD-L1적결합，종이촉사질병진전。
Objective To explore the relationship between the negative co-inhibitor programmed death-1 ( PD-1 ) and the pathogenesis of myasthenia gravis ( MG), by detecting the expression of PD-1 and programmed death ligand-1 ( PD-L1 ) on peripheral blood mononuclear cells (PBMCs) and soluble PD-1 (sPD-1) in plasma from myasthenia gravis patients. Methods Peripheral blood samples were collected from 45 MG patients and 33 healthy persons without prednisone or other immunodepressant treatment during the half year ahead of withdrawal. The expression of PD-1 and PD-L1 on PBMCs were detected using immuno-fluorescence labeling and flow cytometry, and the concentrations of sPD-1 in plasma were measured using an ELISA kit. Results(1) The proportion of CD4+ PD-1 + T cells, as well as CD14+ PD-L1 +monocytes of the MG group was higher than that of the control group. There were no significant differences in the proportion of CD4+ PD-1 + T cells or CD14+ PD-L1 + monocytes in the MG sub-groups between different genders or MG types. While the proportion of CD4+ PD-1 + T cells of the late-onset MG (age ≥40) group was higher than that of the early-onset MG group (age ＜40). And it was higher in the MG patients with thymoma or thymus hyperplasia than that from the MG patients with normal thymus. The proportion of CD14+ PD-L1 +monocytes from the MG patients with thymoma or thymus hyperplasia group decreased obviously compared with that of the patients with normal thymus group; but no difference could be found between the late-onset group and early-onset group. (2)The concentration of sPD-1 in the plasma from the group of MG patients was(6. 92 ±0. 72) ng/ml,which was higher than that of the healthy control group ( (3.28 ±0. 42) ng/ml),even more, it was significantly higher in the early-onset MG group than that of the late-onset MG group,there was a negative correlation( r =-0. 526, P =0. 000) between the age of onset and the concentration of sPD-1. Conclusions The increased expressions of PD-1 on CD4+ T cells and PD-L1 on CD14+ monocytes in MG patients suggested the involvement of the couple of molecules in the pathogenesis of MG. Higher concentration of soluble PD-1 in the plasma of patients with MG suggested that it might disturb the ligation of PD-1 and PD-L1 on T cells and antigen presenting cells, which might result in the abnormal transportation of the negative modulating signal, and accelerate the pathological progress of MG.