Anti-perforin antibody treatment ameliorates experimental crescentic glomerulonephritis in WKY rats

Abstract

The depletion of CD8+ cells has been shown to prevent the initiation and progression of antiglomerular basement membrane (GBM) crescentic glomerulonephritis (GN) in Wistar-Kyoto (WKY) rats. In this study, we asked whether CD8+ cells produce their effects by perforin/granzyme-mediated or by Fas ligand (FasL)-mediated pathways. The glomerular mRNA expression of perforin and granzyme B corresponded with the number of CD8+ cells, whereas that of granzyme A, Fas, and FasL did not. The enhanced mRNA level of perforin and granzyme B was not evident in CD8+-depleted rats. The number of apoptotic cells in the glomeruli was significantly increased at day 3. Perforin mRNA was found in cells infiltrating the glomerulus by in situ hybridization and by using dual-staining immunohistochemistry perforin protein was found in glomerular CD8+ cells. We found that perforin was readily visualized at the inner surface of the glomerular capillaries by immunoelectron microscopy. Based on these results, we treated animals with a perforin antibody in vivo and found that it significantly reduced the amount of proteinuria, frequency of crescentic glomeruli, and the number of glomerular monocytes and macrophages, although the number of glomerular CD8+ cells was not changed. Our results suggest that CD8+ cells play a role in glomerular injury as effector cells in part through a perforin/granzyme-mediated pathway in the anti-GBM WKY rat model of crescentic GN.

Serum-Starved Adipose-Derived Stromal Cells Ameliorate Crescentic GN by Promoting Immunoregulatory Macrophages

“…It has been proposed that glomerular inflammation in anti-GBM GN is a multistep process that includes glomerular recruitment of leukocytes such as neutrophils, monocytes/ macrophages, and lymphocytes, as well as production of proteinases, cytokines/chemokines, and oxygen radicals by inflammatory cells. [23][24][25][26]34,35 Histologic assessment of neutrophils and T cells in diseased glomeruli after TF78 injection revealed a decrease in the accumulation of neutrophils and CD8 + T cells, but not CD4 + T cells, in both HASC-and LASC-treated rats; however, these differences were limited to day 1 (Supplemental Figure 4). Together, these results indicate that during the initial phase, LASCs prevent recruitment of broad leukocyte subsets, including CD8 + T cells, neutrophils, and CD68 + macrophages, and then play an anti-inflammatory role in subsequent phases by promoting an increase in the number of CD163 + macrophages in glomeruli.…”

“…22 This is characteristic of Goodpasture's disease in humans, in which patients may also occasionally present with pulmonary hemorrhage, with dire consequences. Although effector responses by neutrophils, macrophages/monocytes, T lymphocytes, and immune regulation by regulatory T cells and alternatively activated macrophages have been extensively investigated as part of efforts to understand the distinct mechanisms that cause glomerular crescent formation, [23][24][25][26][27][28][29] a potent and feasible therapeutic strategy to inhibit leukocyte effector functions and/or augment their regulatory functions has not emerged for CGN patients. Thus, in this study, we investigated the potential of ASCs to exert immunomodulatory functions in anti-GBM GN.…”

S-Fas Urinary Excretion Helps to Predict the Immunosuppressive Treatment Outcomes in Patients with Proliferative Primary Glomerulonephritis

“…Soluble Fas (sFas) blocks Fas-L and subsequently inhibits apoptosis, causing damage to the glomeruli and usually accelerating the progression of glomerulonephritis (1, 2). The cytotoxic actions mediated via Fas-L may contribute to inflammatory immune responses (3) and Fas-L expression positively correlates with the stage of kidney injury (4). The mRNAs of Fas and Fas-L were also increased in the glomerulus after anti-glomerular basement membrane antibody administration in rats (3).…”

“…The cytotoxic actions mediated via Fas-L may contribute to inflammatory immune responses (3) and Fas-L expression positively correlates with the stage of kidney injury (4). The mRNAs of Fas and Fas-L were also increased in the glomerulus after anti-glomerular basement membrane antibody administration in rats (3). …”

“…Proliferative primary glomerulonephritis comprises mesangial proliferative GN (MesGN), membrano-proliferative GN (MPGN) and immunoglobulin A nephropathy (IgAN), while non-proliferative GN consists of minimal change disease (MCD), focal glomerulosclerosis (FSGS), and membranous nephropathy (MN) (6). In general, proliferative GN may be characterized by glomerular fibrosis - a complicated process initiating cell cytotoxicity and involving many factors, including Fas (1-3, 7, 8). Once induced by Fas an apoptosis helps to prevent local foci of atrophy caused by progressive fibrosis affecting mainly renal tubules (1-3, 7, 8).…”