Autoantibodies to human melanocyte-specific protein pmel17 in the sera of vitiligo patients: a sensitive and quantitative radioimmunoassay (RIA).
In the present study we describe the in vitro transcription-translation of human melanocyte-specific protein Pmel17 cDNA and subsequent use of the resulting 35S-labelled Pmel17 in an RIA to analyse vitiligo sera for the presence of Pmel17 antibodies. Of 53 vitiligo sera examined in the assay, three (5.9%) were found to be positive for Pmel17 antibodies. In contrast, sera from 20 healthy controls, 10 patients with Hashimoto's thyroiditis and 10 patients with Graves' disease (GD) were all negative for Pmel17 antibodies. All three patients positive for Pmel17 antibodies (aged 50-63 years) had had vitiligo of the symmetrical type for > 1 year and all of them also had an associated autoimmune disorder: GD in one and autoimmune hypothyroidism in two. In addition, all three patients had antibodies to the melanogenic enzymes tyrosinase, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2) in their serum. Absorption studies indicated that preincubation with COS-7 cell extract containing expressed Pmel17 absorbed out the immunoreactivity of the three sera positive in the RIA, confirming the anti-Pmel17 reactivity of the sera from these patients. In contrast, COS-7 cell extracts containing either expressed tyrosinase, TRP-1 or TRP-2 did not remove the anti-Pmel17 reactivity of the three sera in the RIA. This lack of cross-reactivity suggests that the humoral response to Pmel17 in these patients is specific and independent of the antibody reactivity to tyrosinase, TRP-1 and TRP-2.
Kemp
E H
EH
Division of Clinical Sciences, Northern General Hospital, University of Sheffield, UK.
Gawkrodger
D J
DJ
Watson
P F
PF
Weetman
A P
AP
D000042:Absorption; D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D001323:Autoantibodies / Q000097:blood* / Q000276:immunology; D019556:COS Cells; D015220:Calcium Channels / Q000235:genetics / Q000276:immunology; D005260:Female; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D008875:Middle Aged; D014442:Monophenol Monooxygenase / Q000235:genetics / Q000276:immunology; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology*; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011506:Proteins; D011863:Radioimmunoassay; D012680:Sensitivity and Specificity; D013462:Sulfur Radioisotopes; D050052:TRPC Cation Channels; D014158:Transcription, Genetic; D014820:Vitiligo / Q000097:blood / Q000276:immunology*; D058951:gp100 Melanoma Antigen
D001323:Autoantibodies; D015220:Calcium Channels; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; C548637:PMEL protein, human; D011506:Proteins; D013462:Sulfur Radioisotopes; D050052:TRPC Cation Channels; C491859:TRPC2 protein, human; C113139:Trp2 protein, vertebrate; D058951:gp100 Melanoma Antigen; C096351:transient receptor potential cation channel, subfamily C, member 1; D014442:Monophenol Monooxygenase