Kemp E H, Waterman E A, Gawkrodger D J, Watson P F, Weetman A P
Division of Clinical Sciences, Northern General Hospital, University of Sheffield, U.K.
J Invest Dermatol. 1999 Aug;113(2):267-71. doi: 10.1046/j.1523-1747.1999.00664.x.
The identification of tyrosinase autoantibodies in some patients with vitiligo has previously been reported. In this study we have determined the B cell epitopes on tyrosinase which are recognized by these autoantibodies. Deletion derivatives of tyrosinase cDNA were constructed and then translated in vitro with the concomitant incorporation of [35S]methionine into the protein products. The 35S-labeled tyrosinase derivatives were subsequently used in radioimmunoassays to investigate the reactivity of sera from five vitiligo patients. The epitope regions identified were: three in a central region of tyrosinase (amino acids 240-255, 289-294, and 295-300) and two others towards the C-terminal end of the protein (amino acids 435-447 and 461-479). Computer analysis of the potential B cell epitopes on tyrosinase revealed that the epitope regions recognized by the vitiligo sera were located in areas predicted to be highly antigenic. In addition, the centrally located antigenic regions (amino acids 289-294 and 295-300) had amino acid sequence homology to both tyrosinase-related protein-1 and -2. Thus, the epitopes on tyrosinase recognized by vitiligo patient sera are heterogeneous and include a region with homology to two related proteins which may explain the cross-reactivity previously noted between these antigens.
此前已有报道称,在一些白癜风患者中发现了酪氨酸酶自身抗体。在本研究中,我们确定了这些自身抗体所识别的酪氨酸酶上的B细胞表位。构建了酪氨酸酶cDNA的缺失衍生物,然后在体外进行翻译,同时将[35S]甲硫氨酸掺入蛋白质产物中。随后,用35S标记的酪氨酸酶衍生物进行放射免疫测定,以研究5例白癜风患者血清的反应性。确定的表位区域为:酪氨酸酶中央区域的三个(氨基酸240 - 255、289 - 294和295 - 300),以及蛋白质C末端的另外两个(氨基酸435 - 447和461 - 479)。对酪氨酸酶上潜在B细胞表位的计算机分析表明,白癜风血清所识别的表位区域位于预测为高度抗原性的区域。此外,位于中央的抗原区域(氨基酸289 - 294和295 - 300)与酪氨酸酶相关蛋白-1和-2均具有氨基酸序列同源性。因此,白癜风患者血清所识别的酪氨酸酶表位是异质性的,包括一个与两种相关蛋白具有同源性的区域,这可能解释了此前所观察到的这些抗原之间的交叉反应性。