Up-regulation of a novel RPE protective protein in experimental uveitis

N. A. Rao, G. S. Wu, Jie Zhang

Research output: Contribution to journalArticle

Abstract

Purpose. Previously, we isolated a novel 79 kDa protein (RPP) secreted by cultured rabbit RPE cells that inhibits superoxide generation by neutrophils and macrophages. This effect is exerted directly on the function of phagocytes, preventing generation of superoxide upon activation. In this study, the in vivo production of RPP during the development of experimental autoimmune uveitis (EAU) was investigated. Methods. The chromatographically pure RPP was utilized to generate polyclonal RPP antibodies (Ab) in both goat and BALB/c mice. The goat Ab was further purified by an ammonium sulfate precipitation and by an Ultrogel AcA 34 size-exclusion chromatography to concentrate the IgG fraction. The Abs were analyzed by ELISA and immunoblot. Experimental uveitis in Lewis rats (EAU) and rabbits (EIU) was induced by S-antigen and by intravitreal injection of lipopolysaccharide (LPS), respectively. Both goat and murine Ab were utilized to detect RPP in the control and inflamed eyes by immunoperoxidase method. Results. The antibodies reacted strongly with purified RPP in ELISA and were blotted to a single band near 79 kDa. The cultured rabbit RPE cells revealed an intense cytoplasmic labeling with the Abs. In control rat eyes, no positive staining was discerned; but an intense and RPE-specific staining was seen in the eyes with EAU at the peak of inflammation. Control rabbit eyes showed a minimal but recognizable level of RPE staining, and the level of staining was intense at the 18-hr post-LPS injection. Conclusion. The RPP, which is a secretory protein of cultured RPE cells, is also produced constitutively at low levels in vivo. This level appears to be up-regulated substantially during inflammation, possibly to suppress the release of oxygen metabolites by stimulated phagocytes, and thus to reduce the tissue damage in uveitis.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996
Externally publishedYes

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Uveitis
Up-Regulation
Goats
Staining and Labeling
Rabbits
Antibodies
Phagocytes
Proteins
Superoxides
Lipopolysaccharides
Enzyme-Linked Immunosorbent Assay
Inflammation
Intravitreal Injections
Ammonium Sulfate
Gel Chromatography
Cultured Cells
Neutrophils
Immunoglobulin G
Macrophages
Oxygen

All Science Journal Classification (ASJC) codes

  • Ophthalmology

Cite this

Up-regulation of a novel RPE protective protein in experimental uveitis. / Rao, N. A.; Wu, G. S.; Zhang, Jie.

In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 3, 15.02.1996.

Research output: Contribution to journalArticle

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