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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 study the relationship between anti-kinectin antibody and Behcet’s disease. Methods RNA was extracted from human laryngeal carcinoma epithelial cells (Hep2 cells) and the nucleotide sequence was amplified by reverse transcription - polymerase chain reaction (RT PCR). The deduced amino acid numbers were 530 ~ 994 (kin m) and 92 0 ~ 1346 (kin 3) and cloned into pET4 2 a (+) vector. The purified fusion protein was purified and then established by enzyme - linked immunosorbent assay (ELISA), and the serum of patients with Behcet’s disease was analyzed. Results The constructed kinectin expression vector could express the antigenic peptide in E. coli. The purified fusion protein was used to establish the ELISA method to detect anti-kinectin antibody. The results showed that the kinectin expression in 46 patients with Behcet’s disease, 118 patients with other autoimmune diseases and The positive rates of kinectin in 51 normal subjects were 6 3% (2/96) for Behcet’s disease, 22% (8/37) for systemic lupus erythematosus and 16% (5/31) for rheumatoid arthritis ), Sjogren’s syndrome 4 1% (11/2 7), mixed connective tissue disease 29% (4/14). Compared with 118 other autoimmune diseases, the detection rate of anti-kinectin antibody was significantly higher in Behcet’s disease (P <0.01). Conclusions Kinectin is a related antigen of Behcet’s disease. Antibody can be detected in other autoimmune diseases, and its significance needs further study.