摘要
目的应用三种表达肽TYR240—300，TYR240—479和TYR检测进展期白癜风患者血清酪氨酸酶IgG抗体，比较三者的抗体检测差异，筛选优势抗原。方法原核表达、纯化三种表达肽TYR240—300，TYR240—479和TYR，Western印迹鉴定纯化蛋白；以纯化的三种表达肽作为抗原，以提取的黑素细胞总蛋白作为阻断抗原，进行阻断ELISA分析；以纯化的三种表达肽作为抗原，检测100例进展期白癜风患者血清中酪氨酸酶IgG抗体，比较三种表达肽的抗体检测阳性率及滴度差异。结果三种表达肽均能被天然的黑素细胞抗原所阻断，100例进展期白癜风患者血清中，抗TYR240—300IgG抗体与抗TYR240—479IgG抗体阳性率分别为32．0％和62．0％，不同型别患者两种抗体检出阳性率差异较大，泛发性（8例）分别为4例和7例，局限性（8例）分别为3例和4例，散在性（48例）分别为31．3％和60．4％，肢端性（19例）分别为20．0％和52．6％，节段型（17例）分别为47．1％和82．4％。40例进展期白癜风患者血清中抗TYR240—479IgG抗体与抗TYRIgG抗体的阳性率相近，分别为62．5％和60．0％。表达肽TYR240—479和TYR检测的进展期白癜风患者血清中IgG抗体最高滴度均为1：320，而表达肽TYR240—300检测的进展期白癜风患者血清中IgG抗体最高滴度均为1：160。结论TYR240—479是用于白癜风患者中酪氨酸酶抗体检测的优势抗原。
Objective To screen superior antigens by comparing the performance of three different expressed epitope peptides of tyrosinase, i.e., TYR240-300, TYR240-479 and TYR in detecting anti-tyrosinase IgG antibodies in the sera of patients with progressive vitiligo. Methods Three epitope peptides, i.e. TYR240-300, TYR240-479 and TYR were previously obtained by prokaryotic expression, purified, and identified with Western blotting. Interception ELISA was carried out with the three epitope peptides as antigens and total protein extracts from primary human melanocytes as interception antigens. Also, purified peptides coated on ELISA plates were used as antigens to detect anti-tyrosinase IgG antibodies in sera from 100 patients with progressive vitiligo, and the positivity and titer of antibodies detected by them were compared. Results All the three epitope peptides could be intercepted by natural melanocyte antigens. The positivity of antibodies against TYR 240-300 and TYR 240-479 varied in patients with different types of vitiligo. In detail, anti-TYR 240-300 antibodies and anti-TYR 240-479 antibodies were detected in 32.0% and 62.0% of the 100 patients, 4 and 7 patients with generalized vitiligo, 3 and 4 patients with localized vitiligo, 15 （31.3%） and 29 （60.4%） patients with sporadic vitiligo, 4 （20.0%） and 10 （52.6%） patients with extremity vitiligo, 8 （47.1%） and 14 （82.4%） patients with segmental vitiligo, respectively. Moreover, the positivity of IgG antibodies against TYR 240-479 and TYR was similar （62.5% vs 62.05%） in 40 tested patients with progressive vitiligo. Besides above, the highest titer of antibody against TYR240-479 and TYR both reached 1 ： 320, and that against TYR240-300 was 1 ： 160. Conelusion TYR240-479 can replace TYR as an antigen for the detection of anti-tyrosinase antibodies in patients with progressive vitiligo.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2009年第10期677-680,共4页
Chinese Journal of Dermatology
基金
国家自然科学基金（30671896,30872280）
卫生部科研基金（EKJ2008-2-033）
浙江省科技厅科技计划（2005C33017）
关键词
白癜风
一元酚单氧酶
免疫显性表位
Vitiligo
Monophenol monooxygenase
Immunodominant epitopes
