The presence of antigen-antibody complexes within the glomerulus is a common initiating mechanism in a wide variety of human glomerulopathies. While the site of formation of these complexes, the route(s) by which they gain access to the glomerulus, their localization, and the nature of the histopathologic reaction they elicit can vary considerably, the central role of the activated leukocyte in initiating and perpetuating pathogenetic events is well-established. A major consequence of PMN/macrophage activation is the release of potent pro-inflammatory oxygenated derivatives of arachidonic acid, in particular prostanoids, thromboxanes, and leukotrienes. Over the past seven years, evidence from our own work and that of other investigators has established enhanced glomerular synthesis, potent glomerular actions, and pathophysiologic relevance for 5- and 15-lipoxygenase (LO) products of arachidonic acid during inflammatory glomerular injury in the rat. More recently, we have obtained evidence that the intraglomerular synthesis of LO products may not be attributed solely to activated leukocytes, but involve the participation of indigenous glomerular cells. This appears to be particularly relevant in the capacity of these cells to effect the transcellular transformation of a key neutrophil/macrophage-derived metabolic intermediate, leukotriene A4, into potent pro-inflammatory eicosanoids. Analysis of the expression of arachidonate LO pathway enzymes in glomerular cells under normal and pathologic conditions is likely to provide significant insight toward understanding their potential roles in glomerular pathophysiology. Utilizing measurements of mRNA in cultured and freshly isolated glomerular cells, in situ hybridization for quantitation of mRNA in fresh tissue sections, and immunocytochemical localization and Western blot analysis of specific enzymes, we will examine the cellular localization and temporal regulation of arachidonate lipoxygenase enzymes gene transcription and translation during the course of glomerular immune injury. We will attempt a correlation of these measurements with end-product synthetic rates and, importantly, with the functional and histomorphologic sequelae of injury. Two representative models of glomerular disease will be studied: anti-glomerular basement membrane antibody-induced glomerulitis, and passive Heymann nephritis. The availability of cDNA clones and specific antibodies for all the major enzymes in these pathways, as well as highly sensitive and accurate analytic techniques for the detection and quantitation of their end-products, provides a unique opportunity to undertake this analysis with a reasonable chance for success.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK043883-04
Application #
3245400
Study Section
Pathology A Study Section (PTHA)
Project Start
1991-06-01
Project End
1996-05-31
Budget Start
1993-06-01
Budget End
1994-05-31
Support Year
4
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Emory University
Department
Type
Schools of Medicine
DUNS #
042250712
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Kelavkar, Uddhav; Wang, Susheng; Badr, Kamal (2002) KU 70/80 lupus autoantigen is the transcription factor induced by interleukins (IL)-13 and -4 leading to induction of 15-lipoxygenase (15-LO) in human cells. Adv Exp Med Biol 507:469-81
Kelavkar, Uddhav; Cohen, Cynthia; Eling, Thomas et al. (2002) 15-lipoxygenase-1 overexpression in prostate adenocarcinoma. Adv Exp Med Biol 507:133-45
Kelavkar, Uddhav; Wang, Susheng; Badr, Kamal (2002) Divergence in intracellular signaling between interleukin-4 (IL-4) and IL-13 in human cells localizes to monomeric/dimeric expression of a transcription factor, the lupus autoantigen 70/80, induced by both cytokines. Adv Exp Med Biol 507:483-9
Kelavkar, U P; Wang, S; Badr, K F (2000) Ku autoantigen (DNA helicase) is required for interleukins-13/-4-induction of 15-lipoxygenase-1 gene expression in human epithelial cells. Genes Immun 1:237-50
Montero, A; Nassar, G M; Uda, S et al. (2000) Reciprocal regulation of LTA(4) hydrolase expression in human monocytes by gamma-interferon and interleukins 4 and 13: potential relevance to leukotriene regulation in glomerular disease. Exp Nephrol 8:258-65
Kelavkar, U P; Cohen, C; Kamitani, H et al. (2000) Concordant induction of 15-lipoxygenase-1 and mutant p53 expression in human prostate adenocarcinoma: correlation with Gleason staging. Carcinogenesis 21:1777-87
Kelavkar, U P; Badr, K F (1999) Effects of mutant p53 expression on human 15-lipoxygenase-promoter activity and murine 12/15-lipoxygenase gene expression: evidence that 15-lipoxygenase is a mutator gene. Proc Natl Acad Sci U S A 96:4378-83
Munger, K A; Montero, A; Fukunaga, M et al. (1999) Transfection of rat kidney with human 15-lipoxygenase suppresses inflammation and preserves function in experimental glomerulonephritis. Proc Natl Acad Sci U S A 96:13375-80
Kelavkar, U; Wang, S; Montero, A et al. (1999) Identification and characterization of an enhancer sequence in the promoter region of human 15-lipoxygenase (15-LO) gene. Adv Exp Med Biol 469:67-74
Kelavkar, U; Wang, S; Montero, A et al. (1998) Human 15-lipoxygenase gene promoter: analysis and identification of DNA binding sites for IL-13-induced regulatory factors in monocytes. Mol Biol Rep 25:173-82

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