Yoshinao Muro1, Kazumitsu Sugiura2, Akira Shiraki3, Norito Ishii4, Takashi Hashimoto4, Masashi Akiyama2. 1. Department of Dermatology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan. Electronic address: ymuro@med.nagoya-u.ac.jp. 2. Department of Dermatology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan. 3. Department of Pulmonary Medicine, Ogaki Municipal Hospital, 4-86 Minaminokawa-cho, Ogaki City, Gifu 503-8502, Japan. 4. Department of Dermatology, Kurume University School of Medicine and Kurume University Institute of Cutaneous Cell Biology, 67 Asahimachi, Kurume, Fukuoka 830-0011, Japan.
Abstract
BACKGROUND: Paraneoplastic pemphigus (PNP) serum preferentially reacts with periplakin and envoplakin, which are plakin family proteins localized to desmosomes and intermediate filaments. Recently, anti-periplakin antibodies were also detected in patients with idiopathic pulmonary fibrosis (IPF). Although previous epitope-mapping studies showed multiple epitopes in each protein, enzyme-linked immunosorbent assays have used several truncated, but not full-length, recombinant proteins. METHODS: This study aimed to produce full-length biotinylated recombinant proteins of periplakin and envoplakin for detection of autoantibodies by immunoprecipitation and ELISA. Serum from a PNP patient who had been confirmed as carrying anti-periplakin and anti-envoplakin antibodies in our previous study was used as a positive control. Sera from 15 patients with IPF were analyzed for both antibodies by immunoprecipitation and by ELISA. RESULTS: The PNP serum reacted strongly with the full-length recombinant proteins in immunoprecipitation and ELISA. Longitudinal serum samples from the PNP patient showed a clear decline of autoantibodies to both periplakin and envoplakin. None of the IPF sera showed both autoantibodies. CONCLUSIONS: We found that the detection of anti-periplakin and anti-envoplakin antibodies using full-length recombinant proteins is useful immunoprecipitation and ELISA.
BACKGROUND:Paraneoplastic pemphigus (PNP) serum preferentially reacts with periplakin and envoplakin, which are plakin family proteins localized to desmosomes and intermediate filaments. Recently, anti-periplakin antibodies were also detected in patients with idiopathic pulmonary fibrosis (IPF). Although previous epitope-mapping studies showed multiple epitopes in each protein, enzyme-linked immunosorbent assays have used several truncated, but not full-length, recombinant proteins. METHODS: This study aimed to produce full-length biotinylated recombinant proteins of periplakin and envoplakin for detection of autoantibodies by immunoprecipitation and ELISA. Serum from a PNP patient who had been confirmed as carrying anti-periplakin and anti-envoplakin antibodies in our previous study was used as a positive control. Sera from 15 patients with IPF were analyzed for both antibodies by immunoprecipitation and by ELISA. RESULTS: The PNP serum reacted strongly with the full-length recombinant proteins in immunoprecipitation and ELISA. Longitudinal serum samples from the PNP patient showed a clear decline of autoantibodies to both periplakin and envoplakin. None of the IPF sera showed both autoantibodies. CONCLUSIONS: We found that the detection of anti-periplakin and anti-envoplakin antibodies using full-length recombinant proteins is useful immunoprecipitation and ELISA.
Authors: Ross Mills; Abhinav Mathur; Lisa M Nicol; Jeremy J Walker; Alexander A Przybylski; Alison C Mackinnon; Sarah E M Howie; William A H Wallace; Ian Dransfield; Nik Hirani Journal: J Immunol Res Date: 2019-04-11 Impact factor: 4.818