摘要
目的 研究抗kinectin抗体与白塞病的关系。方法 从人喉癌上皮细胞 (Hep2细胞 )抽提RNA ,利用反转录 聚合酶链反应 (RT PCR)扩增碱基序列相当于kinectin的氨基酸 5 0 3～ 994(kin m)和氨基酸 92 0～ 1346 (kin 3)两个片段 ,并克隆到pET4 2 a(+)载体中。诱导表达的融合蛋白经纯化后建立酶联免疫吸附试验 (ELISA)方法 ,并对白塞病患者血清进行分析。结果 构建的表达kinectin载体能在大肠杆菌中表达具有抗原性的肽段 ,用纯化的融合蛋白建立检测抗kinectin抗体的ELISA方法 ,对 4 6例白塞病患者、118例其他自身免疫病患者及 5 1名正常人检测显示kinectin抗体阳性率分别为白塞病 6 3% (2 9/ 4 6 ) ,系统性红斑狼疮 2 2 % (8/ 37)、类风湿关节炎 16 % (5 / 31)、干燥综合征 4 1% (11/ 2 7)、混合性结缔组织病 2 9% (4 / 14 )。与 118例其他自身免疫病相比 ,白塞病中抗kinectin抗体的检出率明显升高 (P <0 0 1)。结论 Kinectin是白塞病的相关抗原 ,在其他自身免疫病中也可检测到该抗原抗体 ,其意义有待进一步研究。
Objective To investigate the association of anti-kinectin autoantibody with Behet′s disease.Methods Two fragments of kinectin cloned into prokaryotic expression vector,pET-42a(+) vector,and expressed in BL21(DE3) E.coli.The two fragments kin-m and kin-3 cover kinectin sequence from base 1 579 to 3 050,and 2 829 to 4 107 respectively.The expressed fusion protein were purified by glutathione-S-transferase (GST) purification kits.Using a mixture of two purified fusions,a ELISA was established for detectiing the antibodies.Results Two clones were successfully induced to express fusion protein by isopropyl thiogalactose (IPTG) respectively.The ELISA coated with the mixture of kin-m and kin-3 fusion gave a cut-off value of 0.206 against 95% confidence limit of 51 normal controls,and by this cut-off value,the positive rate of kinectin antibody was 63% in Behet′s disease,16% in RA,22% in SLE,29% in MCTD,and 41% in SS.The prevalence of kinectin antibody did not associate with any clinical manifestation of Behet′s disease but a higher kinectin antibody levels were found in complete Behet′s disease patients.Conclusion Anti-kinectin autoantibody is a Behcet′s disease-related antibody.
出处
《中华风湿病学杂志》
CAS
CSCD
2003年第12期738-741,共4页
Chinese Journal of Rheumatology
基金
国家自然科学基金资助项目 ( 3 0 2 712 2 5 )
上海市高等学校青年科学基金资助项目 ( 0 1QN75 )
