, Volume 26, Issue 2, pp 403–411 | Cite as

CCR2 contributes to the recruitment of monocytes and leads to kidney inflammation and fibrosis development

  • Tarcio Teodoro BragaEmail author
  • Matheus Correa-Costa
  • Reinaldo Correia Silva
  • Mario Costa Cruz
  • Meire Ioshie Hiyane
  • Joao Santana da Silva
  • Katia Regina Perez
  • Iolanda Midea Cuccovia
  • Niels Olsen Saraiva Camara
Original Article


Chemokines are a large family of proteins that, once associated to its receptor on leukocytes, stimulate their movement and migration from blood to tissues. Once in the tissue, immune cells trigger inflammation that, when uncontrolled, leads to fibrosis development. Among the immune cells, macrophages take a special role in fibrosis formation, since macrophage depletion reflects less collagen deposition. The majority of tissue macrophages is derived from monocytes, especially monocytes expressing the chemokine receptor CCR2. Here, we investigated the role of infiltrating CCR2+ cells in the development of fibrosis, and specifically, the dynamic of infiltration of these cells into kidneys under chronic obstructive lesion. Using liposome-encapsulated clodronate, we observed that macrophage depletion culminated in less collagen deposition and reduced chemokines milieu that were released in the damaged kidney after obstructive nephropathy. We also obstructed the kidneys of CCL3−/−, CCR2−/−, CCR4−/−, CCR5−/−, and C57BL/6 mice and we found that among all animals, CCR2−/− mice demonstrated the more robust protection, reflected by less inflammatory and Th17-related cytokines and less collagen formation. Next we evaluated the dynamic of CCR2+/rfp cell infiltration and we observed that they adhere onto the vessels at early stages of disease, culminating in increased recruitment of CCR2+/rfp cells at later stages. On the other hand, CCR2rfp/rfp animals exhibited less fibrosis formation and reduced numbers of recruited cells at later stages. We have experimentally demonstrated that inflammatory CCR2+ cells that reach the injured kidney at initial stages after tissue damage are responsible for the fibrotic pattern observed at later time points in the context of UUO.


CCR2+ monocytes Fibrosis UUO 



We gratefully acknowledge funding provided by CNPq, Capes, and Fapesp (2014/06992-8 and 2012/02270-2).

Compliance with ethical standards

Conflict of interest

None of the authors have any competing interests.


  1. Adegunsoye A, Hrusch CL et al (2016) Skewed lung CCR4 to CCR6 CD4+ T cell ratio in idiopathic pulmonary fibrosis is associated with pulmonary function. Front Immunol 7:516CrossRefPubMedPubMedCentralGoogle Scholar
  2. Bohle A, Wehrmann M et al (1992) The long-term prognosis of the primary glomerulonephritides. A morphological and clinical analysis of 1747 cases. Pathol Res Pract 188(7):908–924CrossRefPubMedGoogle Scholar
  3. Braga TT, Correa-Costa M et al (2012) MyD88 signaling pathway is involved in renal fibrosis by favoring a TH2 immune response and activating alternative M2 macrophages. Mol Med 18:1231–1239CrossRefPubMedPubMedCentralGoogle Scholar
  4. Braga TT, Correa-Costa M et al (2016) Early infiltration of p40IL12(+)CCR7(+)CD11b(+) cells is critical for fibrosis development. Immun Inflamm Dis 4(3):300–314CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bromley SK, Mempel TR et al (2008) Orchestrating the orchestrators: chemokines in control of T cell traffic. Nat Immunol 9(9):970–980CrossRefPubMedGoogle Scholar
  6. Cao Q, Harris DC et al (2015) Macrophages in kidney injury, inflammation, and fibrosis. Physiology (Bethesda) 30(3):183–194Google Scholar
  7. Cassado Ados A, D’Imperio Lima MR et al (2015) Revisiting mouse peritoneal macrophages: heterogeneity, development, and function. Front Immunol 6:225PubMedGoogle Scholar
  8. Chousterman BG, Boissonnas A et al (2016) Ly6Chigh monocytes protect against kidney damage during sepsis via a CX3CR1-dependent adhesion mechanism. J Am Soc Nephrol 27(3):792–803CrossRefPubMedGoogle Scholar
  9. Dey A, Allen J et al (2014) Ontogeny and polarization of macrophages in inflammation: blood monocytes versus tissue macrophages. Front Immunol 5:683PubMedGoogle Scholar
  10. Eddy AA (2000) Molecular basis of renal fibrosis. Pediatr Nephrol 15(3–4):290–301CrossRefPubMedGoogle Scholar
  11. Furuichi K, Wada T et al (2003a) CCR2 signaling contributes to ischemia-reperfusion injury in kidney. J Am Soc Nephrol 14(10):2503–2515CrossRefPubMedGoogle Scholar
  12. Furuichi K, Wada T et al (2003b) Gene therapy expressing amino-terminal truncated monocyte chemoattractant protein-1 prevents renal ischemia-reperfusion injury. J Am Soc Nephrol 14(4):1066–1071CrossRefPubMedGoogle Scholar
  13. Furuichi K, Kaneko S et al (2009) Chemokine/chemokine receptor-mediated inflammation regulates pathologic changes from acute kidney injury to chronic kidney disease. Clin Exp Nephrol 13(1):9–14CrossRefPubMedGoogle Scholar
  14. Geissmann F, Jung S et al (2003) Blood monocytes consist of two principal subsets with distinct migratory properties. Immunity 19(1):71–82CrossRefPubMedGoogle Scholar
  15. Geissmann F, Manz MG et al (2010) Development of monocytes, macrophages, and dendritic cells. Science 327(5966):656–661CrossRefPubMedPubMedCentralGoogle Scholar
  16. Giunti S, Barutta F et al (2010) Targeting the MCP-1/CCR2 System in diabetic kidney disease. Curr Vasc Pharmacol 8(6):849–860CrossRefPubMedGoogle Scholar
  17. Griffith JW, Sokol CL et al (2014) Chemokines and chemokine receptors: positioning cells for host defense and immunity. Annu Rev Immunol 32:659–702CrossRefPubMedGoogle Scholar
  18. Han H, Zhu J et al (2017) Renal recruitment of B lymphocytes exacerbates tubulointerstitial fibrosis by promoting monocyte mobilization and infiltration after unilateral ureteral obstruction. J Pathol 241(1):80–90CrossRefPubMedGoogle Scholar
  19. Kashyap S, Warner GM et al (2016) Blockade of CCR2 reduces macrophage influx and development of chronic renal damage in murine renovascular hypertension. Am J Physiol Renal Physiol 310(5):F372–F384CrossRefPubMedGoogle Scholar
  20. Katagiri D, Hamasaki Y et al (2016) Interstitial renal fibrosis due to multiple cisplatin treatments is ameliorated by semicarbazide-sensitive amine oxidase inhibition. Kidney Int 89(2):374–385CrossRefPubMedGoogle Scholar
  21. Kawamura E, Hisano S et al (2015) Immunohistological analysis for immunological response and mechanism of interstitial fibrosis in IgG4-related kidney disease. Mod Rheumatol 25(4):571–578CrossRefPubMedGoogle Scholar
  22. Kim SM, Lee SH et al (2015) Targeting T helper 17 by mycophenolate mofetil attenuates diabetic nephropathy progression. Transl Res 166(4):375–383CrossRefPubMedGoogle Scholar
  23. Kitagawa K, Wada T et al (2004) Blockade of CCR2 ameliorates progressive fibrosis in kidney. Am J Pathol 165(1):237–246CrossRefPubMedPubMedCentralGoogle Scholar
  24. Kitamoto K, Machida Y et al (2009) Effects of liposome clodronate on renal leukocyte populations and renal fibrosis in murine obstructive nephropathy. J Pharmacol Sci 111(3):285–292CrossRefPubMedGoogle Scholar
  25. Kufareva I, Salanga CL et al (2015) Chemokine and chemokine receptor structure and interactions: implications for therapeutic strategies. Immunol Cell Biol 93(4):372–383CrossRefPubMedPubMedCentralGoogle Scholar
  26. Lefebvre E, Moyle G et al (2016) Antifibrotic effects of the dual CCR2/CCR5 antagonist cenicriviroc in animal models of liver and kidney fibrosis. PLoS One 11(6):e0158156CrossRefPubMedPubMedCentralGoogle Scholar
  27. Mehrotra P, Patel JB et al (2015) Th-17 cell activation in response to high salt following acute kidney injury is associated with progressive fibrosis and attenuated by AT-1R antagonism. Kidney Int 88(4):776–784CrossRefPubMedPubMedCentralGoogle Scholar
  28. Mehrotra P, Collett JA et al (2016) IL-17 mediates neutrophil infiltration and renal fibrosis following recovery from ischemia reperfusion:compensatory role of Natural Killer cells in athymic rats. Am J Physiol Renal Physiol ajprenal 00462:02016Google Scholar
  29. Menezes GB, Rezende RM et al (2008) Differential involvement of cyclooxygenase isoforms in neutrophil migration in vivo and in vitro. Eur J Pharmacol 598(1–3):118–122CrossRefPubMedGoogle Scholar
  30. Mia S, Federico G et al (2015) Impact of AMP-activated protein kinase alpha1 deficiency on tissue Injury following unilateral ureteral obstruction. PLoS One 10(8):e0135235CrossRefPubMedPubMedCentralGoogle Scholar
  31. Nakashima H, Akahoshi M et al (2004) Absence of association between the MCP-1 gene polymorphism and histological phenotype of lupus nephritis. Lupus 13(3):165–167CrossRefPubMedGoogle Scholar
  32. O’Connor T, Borsig L et al (2015) CCL2-CCR2 signaling in disease pathogenesis. Endocr Metab Immune Disord Drug Targets 15(2):105–118CrossRefPubMedGoogle Scholar
  33. Peng X, Zhang J et al (2015a) CX3CL1-CX3CR1 Interaction Increases the Population of Ly6C-CX3CR1hi Macrophages Contributing to Unilateral Ureteral Obstruction-Induced Fibrosis. J Immunol 195(6):2797–2805CrossRefPubMedGoogle Scholar
  34. Peng X, Zhang J et al (2015b) CX3CL1-CX3CR1 interaction increases the population of Ly6C(−)CX3CR1(hi) macrophages contributing to unilateral ureteral obstruction-induced fibrosis. J Immunol 195(6):2797–2805CrossRefPubMedGoogle Scholar
  35. Porubsky S, Schmid H et al (2004) Influence of native and hypochlorite-modified low-density lipoprotein on gene expression in human proximal tubular epithelium. Am J Pathol 164(6):2175–2187CrossRefPubMedPubMedCentralGoogle Scholar
  36. Rot A, von Andrian UH (2004) Chemokines in innate and adaptive host defense: basic chemokinese grammar for immune cells. Annu Rev Immunol 22:891–928CrossRefPubMedGoogle Scholar
  37. Sallusto F, Baggiolini M (2008) Chemokines and leukocyte traffic. Nat Immunol 9(9):949–952CrossRefPubMedGoogle Scholar
  38. Sean Eardley K, Cockwell P (2005) Macrophages and progressive tubulointerstitial disease. Kidney Int 68(2):437–455CrossRefPubMedGoogle Scholar
  39. Segerer S, Nelson PJ (2005) Chemokines in renal diseases. ScientificWorldJournal 5:835–844CrossRefPubMedGoogle Scholar
  40. Segerer S, Nelson PJ et al (2000) Chemokines, chemokine receptors, and renal disease: from basic science to pathophysiologic and therapeutic studies. J Am Soc Nephrol 11(1):152–176PubMedGoogle Scholar
  41. Serbina NV, Pamer EG (2006) Monocyte emigration from bone marrow during bacterial infection requires signals mediated by chemokine receptor CCR2. Nat Immunol 7(3):311–317CrossRefPubMedGoogle Scholar
  42. Sezgin I, Koksal B et al (2011) CCR2 polymorphism in chronic renal failure patients requiring long-term hemodialysis. Intern Med 50(21):2457–2461CrossRefPubMedGoogle Scholar
  43. Shen B, Liu J et al (2016) CCR2 Positive Exosome Released by Mesenchymal Stem Cells Suppresses Macrophage Functions and Alleviates Ischemia/Reperfusion-Induced Renal Injury. Stem Cells Int 2016:1240301CrossRefPubMedPubMedCentralGoogle Scholar
  44. Silva RC, Terra FF et al (2016) Reduced expression of VAChT increases renal fibrosis. Pathophysiology 23(3):229–236CrossRefPubMedGoogle Scholar
  45. Sipos A, Toma I et al (2007) Advances in renal (patho)physiology using multiphoton microscopy. Kidney Int 72(10):1188–1191CrossRefPubMedPubMedCentralGoogle Scholar
  46. Stroo I, Claessen N et al (2015) Deficiency for the chemokine monocyte chemoattractant protein-1 aggravates tubular damage after renal ischemia/reperfusion injury. PLoS One 10(4):e0123203CrossRefPubMedPubMedCentralGoogle Scholar
  47. Sung SA, Jo SK et al (2007) Reduction of renal fibrosis as a result of liposome encapsulated clodronate induced macrophage depletion after unilateral ureteral obstruction in rats. Nephron Exp Nephrol 105(1):e1–e9CrossRefPubMedGoogle Scholar
  48. Trujillo G, O’Connor EC et al (2008) A novel mechanism for CCR4 in the regulation of macrophage activation in bleomycin-induced pulmonary fibrosis. Am J Pathol 172(5):1209–1221CrossRefPubMedPubMedCentralGoogle Scholar
  49. Tsou CL, Peters W et al (2007) Critical roles for CCR2 and MCP-3 in monocyte mobilization from bone marrow and recruitment to inflammatory sites. J Clin Invest 117(4):902–909CrossRefPubMedPubMedCentralGoogle Scholar
  50. Vanhove, T., R. Goldschmeding, et al. (2016). “Kidney fibrosis: origins and interventions.” TransplantationGoogle Scholar
  51. Vielhauer V, Berning E et al (2004) CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome. Kidney Int 66(6):2264–2278CrossRefPubMedGoogle Scholar
  52. Weir MR (2015) CCR2 inhibition: a panacea for diabetic kidney disease? Lancet Diabetes Endocrinol 3(9):666–667CrossRefPubMedGoogle Scholar
  53. Wick G, Grundtman C et al (2013) The immunology of fibrosis. Annu Rev Immunol 31:107–135CrossRefPubMedGoogle Scholar
  54. Wynn TA, Chawla A et al (2013) Macrophage biology in development, homeostasis and disease. Nature 496(7446):445–455CrossRefPubMedPubMedCentralGoogle Scholar
  55. Xia Y, Entman ML et al (2013) CCR2 regulates the uptake of bone marrow-derived fibroblasts in renal fibrosis. PLoS One 8(10):e77493CrossRefPubMedPubMedCentralGoogle Scholar
  56. Yang J, Zhu F et al (2016) Continuous AMD3100 treatment worsens renal fibrosis through regulation of bone marrow derived pro-angiogenic cells homing and T-cell-related inflammation. PLoS ONE 11(2):e0149926CrossRefPubMedPubMedCentralGoogle Scholar
  57. Yuan A, Lee Y et al (2015) Chemokine receptor Cxcr4 contributes to kidney fibrosis via multiple effectors. Am J Physiol Renal Physiol 308(5):F459–F472CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing 2017

Authors and Affiliations

  • Tarcio Teodoro Braga
    • 1
    Email author
  • Matheus Correa-Costa
    • 1
  • Reinaldo Correia Silva
    • 1
  • Mario Costa Cruz
    • 1
  • Meire Ioshie Hiyane
    • 1
  • Joao Santana da Silva
    • 2
  • Katia Regina Perez
    • 3
  • Iolanda Midea Cuccovia
    • 3
  • Niels Olsen Saraiva Camara
    • 1
    • 4
    • 5
  1. 1.Laboratory of Transplantation Immunobiology, Department of Immunology, Institute of Biomedical Sciences IVUniversity of São Paulo (USP)São PauloBrazil
  2. 2.Department of Biochemistry and Immunology, School of Medicine of Ribeirão PretoUniversity of São PauloRibeirão PretoBrazil
  3. 3.Department of BiochemistryInstitute of Chemistry University of São Paulo (USP)São PauloBrazil
  4. 4.Laboratory of Clinical and Experimental Immunology, Nephrology DivisionFederal University of São Paulo (UNIFESP)São PauloBrazil
  5. 5.Renal Pathophysiology Laboratory (LIM16), Faculty of MedicineUniversity of São PauloSão PauloBrazil

Personalised recommendations