A组乙型溶血性链球菌膜蛋白抗原纯化及其在风湿性心脏炎发病中意义的探讨

Purification of protein antigens of group A hemolytic streptococcal membrane and its significance on the onset of rheumatic carditis

目的 探讨纯化A组乙型溶血性链球菌膜蛋白抗原 (A HSMPA)的方法及其在风湿性心脏炎发病免疫过程中的意义。方法 用反胶束萃取法提纯 ,分别检测提纯前后A HSMPA的成分及抗原性 ,再以A HSMPA及粒细胞 /单核细胞集落刺激因子 (GM CSF)分别刺激风湿性心脏炎组、风湿性心脏病静止期组、急性风湿性关节炎组、自身免疫性疾病组、非风湿性心脏病组、链球菌感染相关性疾病组及健康人对照组的外周血淋巴细胞 ,检测其人类白细胞抗原DR亚型 (HLA DR)分子表达量的改变。结果 1 提纯后的A HSMPA基本为蛋白成分 ,提纯前后抗原检测外周淋巴细胞促凝血试验结果无显著差异 ,链球菌壁多糖抗体则差异显著。 2 加入A HSMPA孵育后 ,风湿性心脏炎组HLA DR分子表达量的增加为 (19 5 5± 4 32 )× 10 -6A/cell,对照组为 (12 2 2± 6 2 9)× 10 -6A/cell,各组有显著性差异 ;而经GM CSF刺激后 ,以上各组无明显差别。 3 A HSMPA对风湿性心脏炎患者的刺激作用较GM CSF明显。 4 风湿性心脏炎患者随着症状的好转其HLA DR表达量亦减少。结论 1 反胶束萃取法可提纯A HSMPA并保持其抗原性不变 ,且简便可靠。 2 A HSMPA与HLA DR分子表达量的改变均在风湿性心脏炎发展成为风湿性心脏病过程中起了重要作用。 3 监测A

Objective To investigate the special effect of protein antigens of group A hemolytic streptococcal membrane (A HSMPA)on the onset of rheumatic carditis and its significance on the immune procedure after separated from parietal polysaccharicle antigen Methods Protein antigens of group A hemolytic streptococcal membrane were purified by reversed micelle extraction, and compared with the antigens before purification by hypertensive liquid analysis and ultraviolet spectrophotometer The antigenicities before and after purification were compared with the methods of procoagulant activity and parietal polysaccharicle antibody of serum Then,protein antigens of group A hemolytic streptococcal membrane and GM CSF were used to stimulate the peripheral lymphocytes in groups of normal control, rheumatic carditis, acute rheumatic arthritis, inactive rheumatic heart disease, non rheumatic heart disease, streptococci infection correlative disease and autoimmunity disease The expression level of HLA DR antigen was exanimated by Morris′CELISA Results (1) Parietal polysaccharicle antigens were wiped off after purification (2) After the lymphocytes incubated with protein antigens of group A hemolytic streptococcal membrane, the increase of expression level of HLA DR antigen in the rheumatic carditis group was (19 55±4 32)×10 -6 A/cell, (11 92±4 98)×10 -6 A/cell in inactive rheumatic heart disease group and (11 25±4 18)×10 -6 A/cell in acute rheumatic arthritis group respectively There were significant differences among them After stimulated by GM CSF, the expression level of HLA DR antigen in each group increased But there were no significant differences among them (3) The stimulative effect of protein antigens of group A hemolytic streptococcal membrane was more significant than that of GM CSF (4) The expression level of HLA DR antigens would decrease along with the improvement of rheumatic carditis symptoms But it was still higher than that of normal people Conclusion (1) Protein antigens of group A hemolytic streptococcal membrane would be purified by reversed micelle extraction while parietal polysaccharicle antigens disappear completely Its antigenicity could be maintained (2) Protein antigen of group A hemolytic streptococcal membrane was special stimulus of rheumatic carditis Together with HLA DR, they were nosogenesis of rheumatic carditis (3) The change of HLA DR antigen expression level stimulated by A HSMPA could be one of indexes that decide the treatment period of rheumatic carditis