H Ohguro1, K Ogawa, T Nakagawa. 1. Department of Ophthalmology, Sapporo Medical School of Medicine, Japan.
Abstract
PURPOSE: To characterize retinal autoantigens in patients with cancer-associated retinopathy (CAR). METHODS: The sera of 4 patients with CAR were examined by western blot analysis, and the proteins specifically probed were partially purified from bovine retinas and identified by Edman sequence analysis of the proteolytic peptides. RESULTS: Western blot analysis demonstrated that soluble 23-kDa and 65-kDa proteins were probed by the serum of all the patients. The 23-kDa protein assumed an identical position to that of recoverin when the latter, previously identified as an autoantigen of CAR, was probed by its antibody. This strongly suggested that the 23-kDa antigen and recoverin are identical proteins. After partial purification of the 65-kDa protein from bovine retinas, the corresponding band in two-dimensional gel electrophoresis was cut out and subjected to in-gel digestion by endoproteinase Lys C. Edman sequencing of the proteolytic peptides purified on a high-performance liquid chromatography reverse-phase column identified the 65-kDa protein as the heat shock cognate protein 70 (hsc 70), a member of the heat shock protein (hsp) family involved in protein metabolism as chaperons under both stress and nonstress conditions. To estimate the relationship between the autoimmune responses against recoverin and hsc 70, the maximum serum dilutions required to identify recoverin and hsc 70 on western blot analysis were determined and found to be different among the patients. CONCLUSIONS: These observations suggest that humoral autoimmune reactions against recoverin and hsc 70 might be involved in the pathogenesis of CAR.
PURPOSE: To characterize retinal autoantigens in patients with cancer-associated retinopathy (CAR). METHODS: The sera of 4 patients with CAR were examined by western blot analysis, and the proteins specifically probed were partially purified from bovine retinas and identified by Edman sequence analysis of the proteolytic peptides. RESULTS: Western blot analysis demonstrated that soluble 23-kDa and 65-kDa proteins were probed by the serum of all the patients. The 23-kDa protein assumed an identical position to that of recoverin when the latter, previously identified as an autoantigen of CAR, was probed by its antibody. This strongly suggested that the 23-kDa antigen and recoverin are identical proteins. After partial purification of the 65-kDa protein from bovine retinas, the corresponding band in two-dimensional gel electrophoresis was cut out and subjected to in-gel digestion by endoproteinase Lys C. Edman sequencing of the proteolytic peptides purified on a high-performance liquid chromatography reverse-phase column identified the 65-kDa protein as the heat shock cognate protein 70 (hsc 70), a member of the heat shock protein (hsp) family involved in protein metabolism as chaperons under both stress and nonstress conditions. To estimate the relationship between the autoimmune responses against recoverin and hsc 70, the maximum serum dilutions required to identify recoverin and hsc 70 on western blot analysis were determined and found to be different among the patients. CONCLUSIONS: These observations suggest that humoral autoimmune reactions against recoverin and hsc 70 might be involved in the pathogenesis of CAR.
Authors: Farzin Forooghian; Ian M Macdonald; John R Heckenlively; Elise Héon; Lynn K Gordon; John J Hooks; Barbara Detrick; Robert B Nussenblatt Journal: Am J Ophthalmol Date: 2008-07-30 Impact factor: 5.258
Authors: Monica Goldberg-Murow; Carlos Cedillo-Peláez; Luz Elena Concha-Del-Río; Rashel Cheja-Kalb; María José Salgar-Henao; Eduardo Orozco-Velasco; Héctor Luna-Pastén; Fernando Gómez-Chávez; Antonio Ibarra; Dolores Correa Journal: Front Immunol Date: 2021-05-12 Impact factor: 7.561
Authors: Josianne C Ten Berge; Joost van Rosmalen; Jacolien Vermeer; Cecilia Hellström; Cecilia Lindskog; Peter Nilsson; Ulrika Qundos; Aniki Rothova; Marco W J Schreurs Journal: PLoS One Date: 2016-12-08 Impact factor: 3.240
Authors: Marta Misiuk-Hojło; Maria Ejma; Wojciech A Gorczyca; Stanisław Szymaniec; Danuta Witkowska; Wojciech Fortuna; Ryszard Miedzybrodzki; Jadwiga Rogozińska-Szczepka; Wiesława Bartnik Journal: Arch Immunol Ther Exp (Warsz) Date: 2007-07-23 Impact factor: 4.291