利用Sm抗原模拟表位肽构建狼疮样鼠模型  

Establishment of Systemic Lupus Erythematosus-like Murine Model with Sm Mimotope

作　　者：谢红付[1] 冯浩[2] 曾海燕[1] 李吉[1] 施为[1] 易梅[1] 伍斌[1] 

机构地区：[1]中南大学湘雅医院皮肤科,长沙410008 [2]湖南师范大学第一附属医院皮肤科,长沙410005

出　　处：《中国医学科学院学报》2007年第2期191-195,I0002,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(30571684)~~

摘　　要：目的利用Sm抗原模拟表位肽免疫BALB/C小鼠,构建狼疮样鼠模型。方法用鼠Sm单克隆抗体对噬菌体随机12肽库进行亲和筛选,将筛选的噬菌体阳性克隆行双抗体夹心ELISA检测、DNA序列测定并推导其氨基酸序列,初步确定Sm抗原模拟表位;进而用混合噬菌体阳性克隆皮下免疫BALB/C小鼠,ELISA法检测小鼠血清IgG型抗Sm抗体、抗双链DNA(dsDNA)抗体和抗核抗体(ANA)水平;直接免疫荧光检测小鼠肾脏IgG型免疫复合物的沉积以及HE染色观察小鼠肾脏组织病理损伤情况。结果免疫筛选获得的5个ELISA强阳性的噬菌体克隆的氨基酸序列IR、SQ、PP出现频率增加;实验组小鼠血清中第28天出现Sm抗体、dsDNA抗体、ANA抗体,且滴度不断升高;第33天出现蛋白尿;肾脏可以检测到IgG型免疫复合物沉积及明显的肾脏病理改变。对照组小鼠无明显自身抗体产生和肾脏病变。结论利用Sm抗原噬菌体模拟表位肽可成功构建狼疮样小鼠模型。Objective To establish systemic lupus erythematosus （SLE） -like murine model by immunizing BALB/C mice with Sm mimotope. Methods Sm mimotope was identified by screening a 12-mer random peptide library with monoclonal anti-Smith antibody. Sm mimotope was initially defined with sandwich ELISA, DNA sequencing, and deduced amino acid sequence; and BALB/C mice were subcutaneously injected with mixture phages clones. Sera Sm antibody, anti-double stranded DNA （dsDNA） antibody, and antinuclear antibody （ANA） of mice were detected using direct immunofluorescence; kidney histological changes were examined by HE staining. Results Five randomly selected peptides were sequenced and the amino acid sequences IR, SQ, and PP were detected in a higher frequency. High-titer IgG autoantibodies of dsDNA, Sm, and ANA in the sera of experiment group were detected by ELISA 28 days after having been immunized by Sm mimotope. Proteinuria was detected 33 days later; immune complex and nephritis were observed in kidney specimens. Conclusion SLE-like murine model can be successfully induced by Sm phage mimotope.

关 键 词：系统性红斑狼疮 Sm抗原 噬菌体肽库 模拟表位 

分 类 号：R593.24[医药卫生—内科学]