Cytomegalovirus infection induces expression of 60 KD/Ro antigen on human keratinocytes.
To investigate the effect of human cytomegalovirus (CMV) infection on the expression of Ro autoantigen in human keratinocytes, the binding of anti-Ro peptide antibodies (anti-60 KD/Ro, anti-52 KD/Ro and anti-calreticulin) to cultured human keratinocytes was detected by fixed cell enzyme-linked immunoassay (ELISA), immunofluorescence, flow cytometry (FACS) analysis and immunoblotting. There was a significant increase in the binding of anti-60 KD/Ro antibody but not anti-52 KD/Ro or anti-calreticulin antibody to the surface of cultured keratinocytes at 24 h after CMV infection compared with uninfected cells, by ELISA and immunofluorescence. Surface binding of anti-60 KD/Ro was found in 71.2% (+/- 5.5%) of CMV-infected cells compared with 26.2% (+/- 4.1%) of untreated cells (P < 0.05) by FACS analysis. Similar observations were made with a human serum which contained anti-60 KD/Ro antibodies. Immunoblotting was used to analyse total cellular 60 KD/Ro antigen expression in keratinocytes infected with CMV or without infection. No increase in the intensity of the 60 KD band was found in extracts of the CMV-infected cells, suggesting that the 60 KD/Ro antigen is redistributed from the cytoplasm to the cell surface after viral infection. The effects of CMV infection on cell cultures were compared with those of ultraviolet B (UVB) irradiation. The 60 KD/Ro, 52 KD/Ro and calreticulin were all induced on the UVB-irradiated cell surface but not significant synergistic effect of UVB and CMV was found. This study provides evidence that CMV infection induced 60 KD/Ro antigen expression on the surface of human keratinocytes, suggesting that CMV may play a role in development of skin lesions in systemic lupus erythematosus (SLE).
Zhu
J
J
Department of Medicine, McGill University, Montreal General Hospital Research Institute, Quebec, Canada.
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001324:Autoantigens / Q000032:analysis / Q000096:biosynthesis*; D002460:Cell Line; D003587:Cytomegalovirus / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015603:Keratinocytes / Q000502:physiology* / Q000821:virology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins / Q000032:analysis / Q000096:biosynthesis*; D000099207:SS-A Antigen
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D010446:Peptide Fragments; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C116006:RO60 protein, human