title: Adoptive transfer studies demonstrate that macrophages can induce proteinuria and mesangial cell proliferation.
abstract: BACKGROUND
  Glomerular macrophage accumulation is a feature of proliferative human and experimental glomerulonephritis. However, our understanding of the role of macrophages in the induction of renal injury is based upon indirect evidence from depletion studies, most of which lack specificity for this cell type. Therefore, an adoptive transfer approach was used to directly assess the potential of macrophages to induce renal injury.
  
  
  METHODS
  Accelerated anti-glomerular basement membrane (anti-GBM) disease was induced in rats by immunization with sheep IgG (day -5), followed by administration of sheep anti-rat GBM serum (day 0), with animals killed on day 2. To facilitate the adoptive transfer studies, immunized animals were made leukopenic by cyclophosphamide (CyPh) given on day -2. Bone marrow-derived (BM) or NR8383 macrophages were transferred by tail vein injection 24 hours after injection of anti-GBM serum, with animals killed 3 or 24 hours after transfer.
  
  
  RESULTS
  Pretreatment with CyPh prevented glomerular leukocyte accumulation and completely inhibited proteinuria, glomerular cell proliferation and hypercellularity in accelerated anti-GBM disease. Adoptive transfer led to significant glomerular accumulation of BM or NR8383 macrophages within 3 hours of injection, and this was still evident 24 hours later. Adoptive transfer of BM or NR8383 macrophages induced proteinuria (63 +/- 16 BM vs. 5 +/- 2 mg/24 h CyPh control; P < 0.001), glomerular cell proliferation (5.1 +/- 1.2 BM vs. 0.5 +/- 0.1 PCNA+ cells/gcs CyPh; P < 0.001) and glomerular hypercellularity (51.2 +/- 2.0 BM vs. 41.9 +/- 0.9 nuclei/gcs CyPh; P < 0.001). The degree of renal injury correlated with the number of transferred glomerular macrophages. Two-color immunostaining demonstrated that most glomerular proliferative cell nuclear antigen+ (PCNA+) proliferating cells were OX-7+ mesangial cells. CyPh treatment did not prevent up-regulation of glomerular intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM-1) expression or an increase in urinary monocyte chemoattractant protein-1 (MCP-1) excretion.
  
  
  CONCLUSIONS
  This study provides the first direct evidence that macrophages can induce renal injury in terms of proteinuria and mesangial cell proliferation.
authors.affiliation: Department of Nephrology and Monash University, Clayton, Victoria, Australia.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Ikezumi
authors.forename: Lynette A
authors.initials: LA
authors.lastname: Hurst
authors.forename: Takao
authors.initials: T
authors.lastname: Masaki
authors.forename: Robert C
authors.initials: RC
authors.lastname: Atkins
authors.forename: David J
authors.initials: DJ
authors.lastname: Nikolic-Paterson
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D019867:Anti-Glomerular Basement Membrane Disease / Q000188:drug therapy / Q000276:immunology / Q000473:pathology*; D001854:Bone Marrow Cells; D002455:Cell Division / Q000276:immunology; D002460:Cell Line; D003520:Cyclophosphamide / Q000494:pharmacology; D004195:Disease Models, Animal; D005920:Glomerular Mesangium / Q000276:immunology / Q000473:pathology*; D007074:Immunoglobulin G / Q000494:pharmacology; D007166:Immunosuppressive Agents / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000473:pathology*; D011507:Proteinuria / Q000188:drug therapy / Q000276:immunology / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D007074:Immunoglobulin G; D007166:Immunosuppressive Agents; D003520:Cyclophosphamide