白癜风患者血清抗人黑素浓集素受体-1自身抗体的检测及分析

Detection and Analysis of Autoantibodies of Human Melanin-Concentrating Hormone Receptor 1 in Sera of Patients with Vitiligo

目的检测白癜风患者血清中抗黑素浓集素受体-1(MCHR1)的自身抗体,探讨其在白癜风发病中的意义。方法构建真核表达载体pcDNA3.1/MCHR1,经限制性内切酶双酶切及DNA测序鉴定后,用阳离子脂质体介导转染CHO细胞,经G418筛选,利用ELISA,RTPCR及蛋白印迹法鉴定表达MCHR1蛋白的阳性克隆。采用活细胞ELISA法检测白癜风患者血清抗MCHR1及抗黑素细胞(MC)膜表面抗原的自身抗体。结果白癜风患者血清中抗MCHR1自身抗体阳性率为17.1%,抗MC膜表面抗原的自身抗体阳性率为41.4%,所有抗MCHR1自身抗体阳性的血清抗MC膜表面抗原自身抗体均为阳性,正常人血清中抗MC膜表面抗原及抗MCHR1抗体均为阴性。结论白癜风患者血清中存在抗MCHR1的自身抗体,MCHR1是MC膜表面的白癜风相关抗原。

Objective To detect circulating autoantibodies to human MCHR1 in sera of patients with vitiligo and evaluate its significance in episode of vitiligo.Methods After constructing a eukaryotic expression vector pcDNA3.1/MCHR1 for expression of MCHR1, it was identified by restriction endonuclease digestion and nucleotide sequencing. Then the vector pcDNA3.1/MCHR1 was transfected into CHO cells by mixing with lipofectamine 2000 reagent and screened with G418 to obtain stable expression. The expression of MCHR1 was identified by ELISA,RT-PCR and Western-blot assay. Then the autoantibodies to MCHR1 and melanocyte surface antigens in sera of patients with vitiligo and normal individuals were measured by using a live cell enzyme-linked immunoabsorbent assay. Results Anti-MCHR1 IgG antibodies were present in 17.1% of patients with vitiligo, contrastively anti-melanocyte IgG antibodies were 41.4%. All sera with anti-MCHR1 antibodies were positive to anti-melanocyte. The sera of normal individuals are all negative to anti-MCHR1 and anti-melanocyte.Conclusion There are anti-MCHR1 antibodies in sera of some patients with vitiligo. MCHR1 is one of vitiligo related antigen and it is located on melanocyte membrane.