Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology · 2013
[Detection of serum autoantibodies against premelanosome protein 17 increases in the vitiligo patients].
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Hu Lei, Sun Lijun, Li Yuan, Li Huijin, Liang Daoyan, Sun Jingying, Wang Xin, Hu Jun, Xie Xin
Аннотация
OBJECTIVE: To detect the distribution of autoantibodies against premelanosome protein 17 (Pmel 17) in the sera of vitiligo patients with the recombinant Pmel 17 protein.
METHODS: The cDNA encoding human Pmel 17 was amplified by RT-PCR from the primary melanocytes and was cloned into vector pMD19-T and subsequently the expression vector pGEX-4T-1. After being induced by IPTG, the recombinant Pmel 17 protein was purified by high-performance affinity chromatography. The presence of autoantibodies against Pmel 17 in the sera of vitiligo patients was determined by indirect ELISA coated with recombinant Pmel 17 protein.
RESULTS: The recombinant Pmel 17 plasmids were right as we expected by DNA sequencing. The recombinant Pmel 17 protein was successfully expressed and purified. The indirect ELISA revealed that, as compared with healthy control group (0/100, 0%), positive rate of the serum autoantibodies in the progress vitiligo patients was 22% and positive rate of stable vitiligo patients was only 2%(2/100). The difference of positive rate between patients with progress and stable vitiligo was statistically significant.
CONCLUSION: The recombinant Pmel 17 protein was successfully expressed. The autoantibodies against Pmel 17 is closely related to the severity of vitiligo.
METHODS: The cDNA encoding human Pmel 17 was amplified by RT-PCR from the primary melanocytes and was cloned into vector pMD19-T and subsequently the expression vector pGEX-4T-1. After being induced by IPTG, the recombinant Pmel 17 protein was purified by high-performance affinity chromatography. The presence of autoantibodies against Pmel 17 in the sera of vitiligo patients was determined by indirect ELISA coated with recombinant Pmel 17 protein.
RESULTS: The recombinant Pmel 17 plasmids were right as we expected by DNA sequencing. The recombinant Pmel 17 protein was successfully expressed and purified. The indirect ELISA revealed that, as compared with healthy control group (0/100, 0%), positive rate of the serum autoantibodies in the progress vitiligo patients was 22% and positive rate of stable vitiligo patients was only 2%(2/100). The difference of positive rate between patients with progress and stable vitiligo was statistically significant.
CONCLUSION: The recombinant Pmel 17 protein was successfully expressed. The autoantibodies against Pmel 17 is closely related to the severity of vitiligo.