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Immuno-phenotypic and functional characterization of rabbit pulmonary intravascular macrophages

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Abstract

Pulmonary intravascular macrophages (PIMs) are present in species such as cattle, sheep and horse and promote acute lung inflammation (ALI). Rabbits are often used as a model of ALI but there is controversy about the presence of PIMs in these species. Rabbits were treated with 10 mg/kg of gadolinium chloride intravenously (GC; n = 6) or saline (n = 6) followed by euthanasia at 48 h post-treatment to determine the presence of PIMs. In a subsequent study, rabbits were pre-treated with GC or 0.9 % saline followed by 100 μg/kg of E. coli lipopolysaccharide intravenously 48 h later. Rabbits were euthanized 24 h post-LPS treatment. Light and electron microscopy showed that PIMs attached to the capillary endothelium and were positive for RAM-11 anti-macrophage antibody. While GC treatment induced apoptotic PIMs, there was no difference in the PIM number between control and GC-treated rabbits. Rabbits administered with LPS were 3.5 times more likely to die before the end of the 24-h period than those pre-treated with GC. Lung heterophil accumulation and IL-1β, TNFα and IL-6 mRNA expression were significantly higher in rabbits administered with LPS compared to those administered with GC before the LPS injection. PIMs from the LPS-treated rabbits were positive for TNFα. Lung, BAL and serum IL-8 and MCP-1 expression was not different between LPS rabbits with or without pre-treatment with GC. We conclude that rabbit lungs contain PIMs and that their depletion reduces endotoxin-induced lung inflammation. The presence of PIMs in rabbit lungs may need to be considered while using rabbit to model acute lung injury.

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References

  • Aharonson-Raz K, Singh B (2010) Pulmonary intravascular macrophages and endotoxin-induced pulmonary pathophysiology in horses. Canadian journal of veterinary research. Revue Canadienne de Recherche Veterinaire 74:45–49

    PubMed  CAS  Google Scholar 

  • Altemeier WA, Matute-Bello G, Frevert CW, Kawata Y, Kajikawa O, Martin TR, Glenny RW (2004) Mechanical ventilation with moderate tidal volumes synergistically increases lung cytokine response to systemic endotoxin. Am J Physiol Lung Cell Mol Physiol 287:L533–L542

    Article  PubMed  CAS  Google Scholar 

  • Atwal OS, Saldanha KA (1985) Erythrophagocytosis in alveolar capillaries of goat lung: Ultrastructural properties of blood monocytes. Acta Anat 124:245–254

    Article  PubMed  CAS  Google Scholar 

  • Atwal OS, Minhas KJ, Ferenczy BG, Jassal DS, Milton D, Mahadevappa VG (1989) Morphology of pulmonary intravascular macrophages (PIMs) in ruminants: ultrastructural and cytochemical behavior of dense surface coat. Am J Anat 186:285–299

    Article  PubMed  CAS  Google Scholar 

  • Ben-Shaul V, Sofer Y, Bergman M, Zurovsky Y, Grossman S (1999) Lipopolysaccharide-induced oxidative stress in the liver: comparison between rat and rabbit. Shock 12:288–293

    Article  PubMed  CAS  Google Scholar 

  • Bidani A, Hawkins HK, Wang CZ, Heming TA (1999) Dose dependence and time course of smoke inhalation injury in a rabbit model. Lung 177:111–122

    Article  PubMed  CAS  Google Scholar 

  • Brain JD, Molina RM, Decamp MM, Warner AE (1999) Pulmonary intravascular macrophages: their contribution to the mononuclear phagocyte system in 13 species. Am J Physiol 276:L146–L154

    PubMed  CAS  Google Scholar 

  • Carrasco L, Rodriguez F, Martin de las Mulas J, Sierra MA, Gomez-Villamandos JC, Fernandez A (1991) Pulmonary intravascular macrophages in rabbits experimentally infected with rabbit haemorrhagic disease. J Comp Pathol 105:345–352

    Article  PubMed  CAS  Google Scholar 

  • Chang SW, Ohara N (1994) Chronic biliary obstruction induces pulmonary intravascular phagocytosis and endotoxin sensitivity in rats. J Clin Invest 94:2009–2019

    Article  PubMed  CAS  Google Scholar 

  • Frevert CW, Warner AE (1999) Respiratory distress resulting from acute lung injury in the veterinary patient. J Vet Int Med 6:154–165

    Article  Google Scholar 

  • Frevert CW, Matute-Bello G, Martin TR (2000) Rabbit models of pneumonia, peritoneal sepsis, and lung injury. Methods Mol Biol 138:319–330

    PubMed  CAS  Google Scholar 

  • Gazoni LM, Tribble CG, Zhao MQ, Unger EB, Farrar RA, Ellman PI, Fernandez LG, Laubach VE, Kron IL (2007) Pulmonary macrophage inhibition and inhaled nitric oxide attenuate lung ischemia-reperfusion injury. Ann Thorac Surg 84:247–253

    Article  PubMed  Google Scholar 

  • Gill SS, Suri SS, Janardhan KS, Caldwell S, Duke T, Singh B (2008) Role of pulmonary intravascular macrophages in endotoxin-induced lung inflammation and mortality in a rat model. Respir Res 9:69

    Article  PubMed  Google Scholar 

  • Hildebrand F, Hubbard WJ, Choudhry MA, Frink M, Pape HC, Kunkel SL, Chaudry IH (2006) Kupffer cells and their mediators: the culprits in producing distant organ damage after trauma-hemorrhage. Am J Pathol 169:784–794

    Article  PubMed  CAS  Google Scholar 

  • Horiguchi T, Enzan K, Mitsuhata H, Murata M, Suzuki M (1992) Pulmonary reaction after protamine reversal of heparin in goats and rabbits. Masui 41:1260–1266

    PubMed  CAS  Google Scholar 

  • Horiguchi T, Enzan K, Kawamura K, Suzuki M (1996) Pulmonary responses to heparin–protamine complexes: the effects of age and species. J Appl Physiol 80:56–61

    Article  PubMed  CAS  Google Scholar 

  • Janardhan KS, Appleyard GD, Singh B (2004) Expression of integrin subunits alphav and beta3 in acute lung inflammation. Histochem Cell Biol 121:383–390

    Article  PubMed  CAS  Google Scholar 

  • Janardhan KS, McIsaac M, Fowlie J, Shrivastav A, Caldwell S, Sharma RK, Singh B (2006) Toll like receptor-4 expression in lipopolysaccharide induced lung inflammation. Histol Histopathol 21:687–696

    PubMed  CAS  Google Scholar 

  • Jesmin S, Gando S, Matsuda N, Sakuma I, Kobayashi S, Sakuraya F, Hattori Y (2004) Temporal changes in pulmonary expression of key procoagulant molecules in rabbits with endotoxin-induced acute lung injury: elevated expression levels of protease-activated receptors. Thromb Haemost 92:966–979

    PubMed  CAS  Google Scholar 

  • Kajikawa O, Goodman RB, Johnson MC 2nd, Konishi K, Martin TR (1996) Sensitive and specific immunoassays to detect rabbit IL-8 and MCP-1: cytokines that mediate leukocyte recruitment to the lungs. J Immunol Methods 197:19–29

    Article  PubMed  CAS  Google Scholar 

  • Kawashima M, Kuwamura M, Takeya M, Yamate J (2004) Morphologic characteristics of pulmonary macrophages in cetaceans: particular reference to pulmonary intravascular macrophages as a newly identified type. Vet Pathol 41:682–686

    Article  PubMed  CAS  Google Scholar 

  • Kuebler WM (2005) Inflammatory pathways and microvascular responses in the lung. Pharmacol Rep 57(Suppl):196–205

    PubMed  Google Scholar 

  • Kurahashi K, Kajikawa O, Sawa T, Ohara M, Gropper MA, Frank DW, Martin TR, Wiener-Kronish KP (1999) Pathogenesis of septic shock in Pseudomonas aeruginosa pneumonia. J Clin Invest 104:743–750

    Article  PubMed  CAS  Google Scholar 

  • Kurahashi K, Sawa T, Ota M, Kajikawa O, Hong K, Martin TR, Wiener-Kronish JP (2009) Depletion of phagocytes in the reticuloendothelial system causes increased inflammation and mortality in rabbits with Pseudomonas aeruginosa pneumonia. Am J Physiol Lung Cell Mol Physiol 296:L198–L209

    Article  PubMed  CAS  Google Scholar 

  • Longworth KE (1997) The comparative biology of pulmonary intravascular macrophages. Front Biosci 2:232–241

    Google Scholar 

  • Mathison JC, Ulevitch RJ (1979) The clearance, tissue distribution, and cellular localization of intravenously injected lipopolysaccharide in rabbits. J Immunol 123:2133–2143

    PubMed  CAS  Google Scholar 

  • Mathison JC, Ulevitch RJ (1981) In vivo interaction of bacterial lipopolysaccharide (LPS) with rabbit platelets: modulation by C3 and high density lipoproteins. J Immunol 126:1575–1580

    PubMed  CAS  Google Scholar 

  • Matute-Bello G, Frevert CW, Martin TR (2008) Animal models of acute lung injury. Am J Physiol Lung Cell Mol Physiol 295:L379–L399

    Article  PubMed  CAS  Google Scholar 

  • Molina RM, Brain JD (2007) In vivo comparison of cat alveolar and pulmonary intravascular macrophages: phagocytosis, particle clearance, and cytoplasmic motility. Exp Lung Res 33:53–70

    Article  PubMed  CAS  Google Scholar 

  • Parbhakar OP, Duke T, Townsend HG, Singh B (2004) Immunophenotypic characterization and depletion of pulmonary intravascular macrophages of horses. Vet Res 35:39–51

    Article  PubMed  Google Scholar 

  • Parbhakar OP, Duke T, Townsend HG, Singh B (2005) Depletion of pulmonary intravascular macrophages partially inhibits lipopolysaccharide-induced lung inflammation in horses. Vet Res 36:557–569

    Article  PubMed  Google Scholar 

  • Poli-de-Figueiredo LF, Garrido AG, Nakagawa N, Sannomiya P (2008) Experimental models of sepsis and their clinical relevance. Shock 30(Suppl 1):53–59

    Article  PubMed  CAS  Google Scholar 

  • Ramadori G, Moriconi F, Malik I, Dudas J (2008) Physiology and pathophysiology of liver inflammation, damage and repair. J Physiol Pharmacol 59(Suppl 1):107–117

    PubMed  Google Scholar 

  • Reutershan J, Basit A, Galkina EV, Ley K (2005) Sequential recruitment of neutrophils into lung and bronchoalveolar lavage fluid in LPS-induced acute lung injury. Am J Physiol Lung Cell Mol Physiol 289:L807–L815

    Article  PubMed  CAS  Google Scholar 

  • Ribble CS, Meek AH, Janzen ED, Jim GK (1995) Effect of time of year, weather, and the pattern of auction market sales on fatal fibrinous pneumonia (shipping fever) in calves in a larger feedlot in Alberta (1985–1988). Can J Vet Res 59:167–172

    PubMed  CAS  Google Scholar 

  • Schneberger D, Aharonson-Raz K, Singh B (2012) Pulmonary intravascular macrophages and lung health: what are we missing? Am J Physiol Lung Cell Mol Physiol 302:L498–L503

    Article  PubMed  CAS  Google Scholar 

  • Schneider T, Issekutz AC (1996) Quantitation of eosinophil and neutrophil infiltration into rat lung by specific assays for eosinophil peroxidase and myeloperoxidase. Application in a Brown Norway rat model of allergic pulmonary inflammation. J Immunol Methods 198:1–14

    Article  PubMed  CAS  Google Scholar 

  • Shimada H, Hasegawa N, Koh H, Tasaka S, Shimizu M, Yamada W, Nishimura T, Amakawa K, Kohno M, Sawafuji M, Nakamura K, Fujishima S, Yamaguchi K, Ishizaka A (2006) Effects of initial passage of endotoxin through the liver on the extent of acute lung injury in a rat model. Shock 26:311–315

    Article  PubMed  CAS  Google Scholar 

  • Singh B, de la Concha-Bermejillo A (1998) Gadolinium chloride removes pulmonary intravascular macrophages and curtails the degree of ovine lentivirus-induced lymphoid interstitial pneumonia. Int J Exp Pathol 79:151–162

    PubMed  CAS  Google Scholar 

  • Singh B, Pearce JW, Gamage LN, Janardhan K, Caldwell S (2004) Depletion of pulmonary intravascular macrophages inhibits acute lung inflammation. Am J Physiol Lung Cell Mol Physiol 286:L363–L372

    Article  PubMed  CAS  Google Scholar 

  • Sone Y, Serikov VB, Staub NC (1999) Intravascular macrophage depletion attenuates endotoxin lung injury in anesthetized sheep. J Appl Physiol 87:1354–1359

    PubMed  CAS  Google Scholar 

  • Staub NC (1994) Pulmonary intravascular macrophages. Ann Rev Physiol 56:47–67

    Article  CAS  Google Scholar 

  • Warner AE, Brain JD (1990) The cell biology and pathogenic role of pulmonary intravascular macrophages. Am J Physiol 258:L1–L12

    PubMed  CAS  Google Scholar 

  • Warner AE, Barry BA, Brain JD (1986) Pulmonary intravascular macrophages in sheep: Morphology and function of a novel constituent of the mononuclear phagocyte system. Lab Invest 55:276–288

    PubMed  CAS  Google Scholar 

  • Werners AH, Bull S, Fink-Gremmels J (2005) Endotoxaemia: a review with implications for the horse. Equine Vet J 37:371–383

    Article  PubMed  CAS  Google Scholar 

  • Winkler GC (1988) Pulmonary intravascular macrophages in domestic animal species: Review of structural and functional properties. Am J Anat 181:217–234

    Article  PubMed  CAS  Google Scholar 

  • Winkler GC, Cheville NF (1985) Monocytic origin and postnatal mitosis of intravascular macrophages in the porcine lung. J Leukoc Biol 38:471–480

    PubMed  CAS  Google Scholar 

  • Winkler GC, Cheville NF (1987) Postnatal colonization of porcine lung capillaries by intravascular macrophages: An ultrastructural, morphometric analysis. Microvasc Res 33:224–232

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

The research was supported through a grant to Dr. Duke-Novakovski and Dr. Singh from the Equine Health Research Fund of the Western College of Veterinary Medicine and a grant from the Natural Science and Engineering Research Council of Canada to Dr. Baljit Singh.

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Authors and Affiliations

  1. Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan, S7N 5B4, Canada

    Tanya Duke-Novakovski, Sarabjeet Singh-Suri, Sarah Caldwell, Chandarshekhar Charavaryamath & Baljit Singh

  2. Medical Research Service of the VA Puget Sound Health Center, Seattle, WA, USA

    Osamu Kajikawa

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  1. Tanya Duke-Novakovski
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  2. Sarabjeet Singh-Suri
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  3. Osamu Kajikawa
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  4. Sarah Caldwell
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  5. Chandarshekhar Charavaryamath
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  6. Baljit Singh
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Correspondence to Baljit Singh.

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Duke-Novakovski, T., Singh-Suri, S., Kajikawa, O. et al. Immuno-phenotypic and functional characterization of rabbit pulmonary intravascular macrophages. Cell Tissue Res 351, 149–160 (2013). https://doi.org/10.1007/s00441-012-1509-2

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  • DOI: https://doi.org/10.1007/s00441-012-1509-2

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