Increased expression of the pro-apoptotic ATP-sensitive P2X7 receptor in experimental and human glomerulonephritis.

BACKGROUND The involvement of IL-1beta and other pro-inflammatory cytokines in most forms of glomerulonephritis is now well established. The P2X(7) receptor, an ATP-sensitive P2X receptor, functions not only as a non-selective cation channel, but it is also involved in the rapid processing and release of IL-1beta, apoptosis and necrotic cell death. Therefore, we wanted to investigate if expression of this receptor is altered in the glomeruli of rodent models of glomerulonephritis. METHODS P2X(7) receptor protein expression was investigated using immunohistochemistry, and apoptosis was assessed using the TUNEL assay and caspase-3 immunostaining. Real-time PCR with gene-specific primers was used to detect P2X(7), IL-1beta, p53, bax and bcl-2 mRNA expression. RESULTS Although the levels of the P2X(7) receptor protein in mouse kidney are normally very low, or undetectable, we detected an increase in glomerular expression of this receptor and an increase in glomerular apoptotic cells in a mouse model of accelerated nephrotoxic nephritis. We also observed increased glomerular and tubular expression of the P2X(7) receptor protein in renal biopsy tissue of patients with autoimmune-related glomerulonephritis. Furthermore, P2X(7) receptor mRNA increased in the kidneys of a rat model of proliferative glomerulonephritis and this coincided with the onset of proteinuria. We also observed increased mRNA expression of Il-1beta and the pro-apoptotic markers p53 and bax, but not of anti-apoptotic bcl-2. CONCLUSION Although there is an association between expression of the pro-inflammatory and pro-apoptotic P2X(7) receptor and glomerulonephritis in these rodent models, and in at least one form of human glomerulonephritis, the underlying relationship and its functional significance remain to be explored.

[1]  J. Kanellopoulos,et al.  P2Z/P2X7receptor-dependent apoptosis of dendritic cells. , 1999, American journal of physiology. Cell physiology.

[2]  A. Steinkasserer,et al.  Differential expression of macrophage inflammatory protein-2 and monocyte chemoattractant protein-1 in experimental glomerulonephritis. , 1996, Kidney international.

[3]  C. Pusey,et al.  Development of scarring and renal failure in a rat model of crescentic glomerulonephritis. , 1999, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[4]  Denise Feighan,et al.  P2X7-Like Receptor Activation in Astrocytes Increases Chemokine Monocyte Chemoattractant Protein-1 Expression via Mitogen-Activated Protein Kinase , 2001, The Journal of Neuroscience.

[5]  M. Walport,et al.  Both Fcgamma receptor I and Fcgamma receptor III mediate disease in accelerated nephrotoxic nephritis. , 2003, American Journal of Pathology.

[6]  F. Di Virgilio.  The P2Z purinoceptor: an intriguing role in immunity, inflammation and cell death. , 1995, Immunology today.

[7]  K. Poelstra,et al.  Modulation of anti-Thy1 nephritis in the rat by adenine nucleotides. Evidence for an anti-inflammatory role for nucleotidases. , 1992, Laboratory investigation; a journal of technical methods and pathology.

[8]  C. Pusey,et al.  Prevention and treatment of experimental crescentic glomerulonephritis by blocking tumour necrosis factor-alpha. , 2001, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[9]  E. Kawashima,et al.  Tissue distribution of the P2X7 receptor , 1997, Neuropharmacology.

[10]  C. Gabel,et al.  ATP Acts as an Agonist to Promote Stimulus-Induced Secretion of IL-1β and IL-18 in Human Blood , 2000, The Journal of Immunology.

[11]  A. Woolf,et al.  The P2X7 ATP receptor modulates renal cyst development in vitro. , 2004, Biochemical and biophysical research communications.

[12]  G. Burnstock,et al.  The Pattern of Distribution of Selected ATP-Sensitive P2 Receptor Subtypes in Normal Rat Kidney: An Immunohistological Study , 2003, Cells Tissues Organs.

[13]  P. Tipping,et al.  Contributions of IL-1β and IL-1α to Crescentic Glomerulonephritis in Mice , 2004 .

[14]  P. Tipping,et al.  Contributions of IL-1beta and IL-1alpha to crescentic glomerulonephritis in mice. , 2004, Journal of the American Society of Nephrology : JASN.

[15]  N. Rothwell,et al.  Priming of Macrophages with Lipopolysaccharide Potentiates P2X7-mediated Cell Death via a Caspase-1-dependent Mechanism, Independently of Cytokine Production* , 2002, The Journal of Biological Chemistry.

[16]  R. Atkins,et al.  Intrinsic renal cells are the major source of interleukin-1 beta synthesis in normal and diseased rat kidney. , 1997, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[17]  C. Dinarello.  Interleukin-1 beta, interleukin-18, and the interleukin-1 beta converting enzyme. , 1998, Annals of the New York Academy of Sciences.

[18]  Beverly H. Koller,et al.  Altered Cytokine Production in Mice Lacking P2X7Receptors* , 2001, The Journal of Biological Chemistry.

[19]  Kazuhide Inoue,et al.  Extracellular ATP Triggers Tumor Necrosis Factor‐α Release from Rat Microglia , 2000 .

[20]  G. Burnstock,et al.  The P2X7 ATP Receptor in the Kidney: A Matter of Life or Death? , 2005, Nephron Experimental Nephrology.

[21]  G. Dubyak,et al.  P2X7 Receptor-Dependent Blebbing and the Activation of Rho-Effector Kinases, Caspases, and IL-1β Release1 , 2003, The Journal of Immunology.

[22]  F. Autschbach,et al.  Podocytes are the major source of IL-1α and IL-1β in human glomerulonephritides , 1997 .

[23]  C. Dinarello.  Interleukin‐1β, Interleukin‐18, and the Interleukin‐1β Converting Enzyme a , 1998 .

[24]  M. Walport,et al.  Both Fcγ Receptor I and Fcγ Receptor III Mediate Disease in Accelerated Nephrotoxic Nephritis , 2003 .

[25]  G. Burnstock,et al.  Glomerular expression of the ATP-sensitive P2X receptor in diabetic and hypertensive rat models. , 2004, Kidney international.

[26]  R. North,et al.  Epithelial Membrane Proteins Induce Membrane Blebbing and Interact with the P2X7 Receptor C Terminus* , 2002, The Journal of Biological Chemistry.

[27]  P. Tipping,et al.  Glomerular interleukin 1 production is dependent on macrophage infiltration in anti-GBM glomerulonephritis. , 1991, Kidney international.

[28]  D. Rampe,et al.  P2X7 receptor modulation of β-amyloid- and LPS-induced cytokine secretion from human macrophages and microglia , 2004, Journal of Neuroimmunology.

[29]  B. Yang,et al.  Apoptosis and caspase-3 in experimental anti-glomerular basement membrane nephritis. , 2001, Journal of the American Society of Nephrology : JASN.

[30]  R. Atkins,et al.  Suppression of experimental crescentic glomerulonephritis by the interleukin-1 receptor antagonist. , 1993, Kidney international.

[31]  G. Burnstock,et al.  Induction of proliferation and apoptotic cell death via P2Y and P2X receptors, respectively, in rat glomerular mesangial cells. , 2000, Kidney international.

[32]  Y. Nakata,et al.  Extracellular ATP triggers tumor necrosis factor-alpha release from rat microglia. , 2000, Journal of neurochemistry.

[33]  T. Steinberg,et al.  Extracellular ATP induces a large nonselective conductance in macrophage plasma membranes. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[34]  I. P. Chessell,et al.  Pharmacological characterization of ATP‐ and LPS‐induced IL‐1β release in human monocytes , 1999 .

[35]  E. Kawashima,et al.  The Cytolytic P2Z Receptor for Extracellular ATP Identified as a P2X Receptor (P2X7) , 1996, Science.

[36]  G. Dubyak,et al.  Essential role for Ca2+ in regulation of IL-1beta secretion by P2X7 nucleotide receptor in monocytes, macrophages, and HEK-293 cells. , 2003, American journal of physiology. Cell physiology.

[37]  F. Autschbach,et al.  Podocytes are the major source of IL-1 alpha and IL-1 beta in human glomerulonephritides. , 1997, Kidney international.

[38]  C. Pusey,et al.  Prevention and treatment of experimental crescentic glomerulonephritis by blocking tumour necrosis factor-alpha. , 2001, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[39]  L. Welt,et al.  Diseases of the Kidney , 1905, The Hospital.

[40]  F. Di Virgilio,et al.  Extracellular ATP triggers IL-1 beta release by activating the purinergic P2Z receptor of human macrophages. , 1997, Journal of immunology.

[41]  C. Gabel,et al.  Absence of the P2X7 Receptor Alters Leukocyte Function and Attenuates an Inflammatory Response , 2002, The Journal of Immunology.

[42]  K. Matsumoto,et al.  Production of interleukin 1 in glomerular cell cultures from patients with rapidly progressive crescentic glomerulonephritis. , 1988, American journal of nephrology.

[43]  G. Dubyak,et al.  Essential role for Ca 2 (cid:1) in regulation of IL-1 (cid:2) secretion by P2X 7 nucleotide receptor in monocytes, macrophages, and HEK-293 cells , 2003 .

[44]  R. North,et al.  Kinetics of cell lysis, dye uptake and permeability changes in cells expressing the rat P2X7 receptor , 1999, The Journal of physiology.

[45]  B. Gomperts,et al.  ATP induces nucleotide permeability in rat mast cells , 1979, Nature.

[46]  P. Tipping,et al.  Leukocyte-derived interleukin-1beta interacts with renal interleukin-1 receptor I to promote renal tumor necrosis factor and glomerular injury in murine crescentic glomerulonephritis. , 2004, The American journal of pathology.