• 1.

    Harris NB, Barletta RG. Mycobacterium avium subsp. paratuberculosis in veterinary medicine. Clin Microbiol Rev 2002;14:489512.

  • 2.

    Clarke CJ. The pathology and pathogenesis of paratuberculosis in ruminants and other species. J Comp Pathol 1997;116:217261.

  • 3.

    Waters WR, Miller JM, Palmer MV, et al. Early induction of humoral and cellular immune responses during experimental Mycobacterium avium subsp. paratuberculosis infection in calves. Infect Immun 2003;71:51305138.

    • Search Google Scholar
    • Export Citation
  • 4.

    Ernst JD. Macrophage receptors for Mycobacterium tuberculosis. Infect Immun 1998;66:12771281.

  • 5.

    Underhill DM, Ozinsky A. Phagocytosis of microbes: complexity in action. Annu Rev Immunol 2002;20:825852.

  • 6.

    Schlesinger LS, Bellinger-Kawahara CG, Payne NR, et al. Phagocytosis of Mycobacterium tuberculosis is mediated by human monocyte complement receptors and component C3. J Immunol 1990;144:27712780.

    • Search Google Scholar
    • Export Citation
  • 7.

    Schlesinger LS. Macrophage phagocytosis of virulent but not attenuated strains of Mycobacterium tuberculosis is mediated by mannose receptors in addition to complement receptors. J Immunol 1993;150:29202930.

    • Search Google Scholar
    • Export Citation
  • 8.

    Peterson PK, Gekker G, Hu S, et al. CD14 receptor-mediated uptake of nonopsonized Mycobacterium tuberculosis by human microglia. Infect Immun 1995;63:15981602.

    • Search Google Scholar
    • Export Citation
  • 9.

    Woo S, Sotos J, Hart AP, et al. Bovine monocytes and a macrophage cell line differ in their ability to phagocytose and support the intracellular survival of Mycobacterium avium subsp. paratuberculosis. Vet Immunol Immumopathol 2006;110:109120.

    • Search Google Scholar
    • Export Citation
  • 10.

    Souza CD, Evanson OA, Weiss DJ. Mitogen activated protein kinase (p38) pathway is an important component of the anti-inflammatory response in Mycobacterium avium subsp. paratuberculosisinfected bovine monocytes. Microb Pathog 2006;41:5966.

    • Search Google Scholar
    • Export Citation
  • 11.

    Souza CD, Evanson OA, Weiss DJ. Regulation by Jun N-terminal kinase/stress activated protein kinase of cytokine expression in Mycobacterium avium subsp paratuberculosis–infected bovine monocytes. Am J Vet Res 2006;67:17601765.

    • Search Google Scholar
    • Export Citation
  • 12.

    Schorey JS, Cooper AM. Macrophage signaling upon mycobacterial infection: the MAP kinases lead the way. Cell Microbiol 2003;5:133142.

  • 13.

    Wells SJ, Faaberg KS, Wees C, et al. Evaluation of a rapid fecal PCR test for detection of Mycobacterium avium subsp. paratuberculosis in dairy cattle. Clin Vaccine Immunol 2006;13:11251130.

    • Search Google Scholar
    • Export Citation
  • 14.

    Weiss DJ, Evanson OA, Moritz A, et al. Differential response of bovine macrophages to Mycobacterium avium subsp.paratuberculosis and Mycobacterium avium subsp. avium. Infect Immun 2002;70:55565561.

    • Search Google Scholar
    • Export Citation
  • 15.

    Stabel JR, Stabel TJ. Immortalization and characterization of bovine peritoneal macrophages transfected with SV40 plasmid DNA. Vet Immun Immunopathol 1995;45:211220.

    • Search Google Scholar
    • Export Citation
  • 16.

    Sager H, Davis WC, Jungi TW. Bovine monocytoid cells transformed to proliferate cease to exhibit lineage-specific functions. Vet Immunol Immunopathol 1999;68:113130.

    • Search Google Scholar
    • Export Citation
  • 17.

    Sohn EJ, Paape MJ, Peters RR, et al. The production and characterization of anti-bovine CD14 monoclonal antibodies. Vet Res 2004;35:597608.

    • Search Google Scholar
    • Export Citation
  • 18.

    Sabroe I, Jones EC, Usher LR, et al. Toll-like receptors (TLR)2 and TLR4 in human peripheral blood granulocytes: a critical role for monocytes in leukocyte lipopolysaccharide responses. J Immunol 2002;168:47014710.

    • Search Google Scholar
    • Export Citation
  • 19.

    Diaz-Silvestre H, Espinosa-Cueto P, Sanchez-Gonzalez A, et al. The 19-kDa antigen of Mycobacterium tuberculosis is a major adhesin that binds the mannose receptor of THP-1 monocytic cells and promotes phagocytosis of mycobacteria. Microb Pathog 2005;39:97107.

    • Search Google Scholar
    • Export Citation
  • 20.

    Fattorossi A, Nisini R, Pizzolo, et al. New, simple flow cytometry technique to discriminate between internalized and membranebound particles in phagocytosis. Cytometry 1989;10:320325.

    • Search Google Scholar
    • Export Citation
  • 21.

    Nigou J, Zelle-Rieser C, Gilleron M, et al. Mannosylated lipoarabinomannans inhibit IL-12 production by human dendritic cells: evidence for a negative signal delivered through the mannose receptor. J Immunol 2001;166:74777485.

    • Search Google Scholar
    • Export Citation
  • 22.

    Khoo K, Tang J, Chatterjee D. Variation in mannose-capped terminal arabinan motifs of lipoarabinomannans from clinical isolates of Mycobacterium tuberculosis and Mycobacterium avium complex. J Biol Chem 2001;276:38633871.

    • Search Google Scholar
    • Export Citation
  • 23.

    Khanna KV, Choi CS, Gekker G, et al. Differential infection of porcine alveolar macrophage subpopulations by nonopsonized Mycobacterium bovis involves CD14 receptors. J Leukoc Biol 1996;60:214220.

    • Search Google Scholar
    • Export Citation
  • 24.

    Pugin J, Heumann ID, Tomasz A, et al. CD14 is a pattern recognition receptor. Immunity 1994;1:509516.

  • 25.

    Zimmerli S, Edwards S, Ernst JD. Selective receptor blockade during phagocytosis does not alter the survival and growth of Mycobacterium tuberculosis in human macrophages. Am J Respir Biol 1996;15:760770.

    • Search Google Scholar
    • Export Citation

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Cell membrane receptors on bovine mononuclear phagocytes involved in phagocytosis of Mycobacterium avium subsp paratuberculosis

Cleverson D. Souza DVM, MS1, Oral A. Evanson BS2, Srinand Sreevatsan MVSc, MPH, PhD3, and Douglas J. Weiss DVM, PhD4
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  • 1 Department of Veterinary and Biomedical Science, College of Veterinary Medicine, University of Minnesota, Saint Paul, MN 55108.
  • | 2 Department of Veterinary and Biomedical Science, College of Veterinary Medicine, University of Minnesota, Saint Paul, MN 55108.
  • | 3 Department of Veterinary Population Sciences, College of Veterinary Medicine, University of Minnesota, Saint Paul, MN 55108.
  • | 4 Department of Veterinary and Biomedical Science, College of Veterinary Medicine, University of Minnesota, Saint Paul, MN 55108.

Abstract

Objective—To determine cell membrane receptors involved in phagocytosis of Mycobacterium avium subsp paratuberculosis (MAP) organisms.

Sample Population—Monocytes were obtained from healthy adult Holstein dairy cows that were test negative for MAP infection on the basis of bacteriologic culture of feces and serologic test results.

Procedures—Monocytes or bovine macrophage cell line (BoMac) cells were incubated with MAP organisms for 30, 60, or 120 minutes with or without inhibitors of integrins, CD14, or mannose receptors. Phagocytosis was evaluated by light microscopy or by flow cytometry. CD11a/CD18, CD11b, and CD14 expression on monocytes and BoMac cells was evaluated by use of flow cytometry.

Results—Monocytes and BoMac cells rapidly phagocytized MAP organisms. However, compared with BoMac cells, monocytes had a greater total capacity to phagocytize MAP organisms. Addition of neutralizing anti-integrin antibodies (anti-CD11a/CD18 and anti-CD11b) substantially inhibited phagocytosis by monocytes during the first 60 minutes of incubation with MAP organisms, but were less effective at 120 minutes of incubation. Anti-CD11a/CD18 and anti-CD11b antibodies were less effective in inhibiting phagocytosis by BoMac cells. Addition of inhibitors of CD14 or mannose receptors also inhibited phagocytosis of MAP by monocytes. Addition of a combination of integrin and mannose inhibitors had an additive effect in reducing phagocytosis, but addition of integrin and CD14 inhibitors did not have an additive effect.

Conclusions and Clinical Relevance—Multiple receptors are involved in phagocytosis of MAP organisms. Although CD11/CD18 receptors appear to be the major receptors used by MAP at early time points, mannose receptors and CD14 also contribute substantially to phagocytosis.

Contributor Notes

Supported in part by a grant from the USDA-NRI-05-010447. Dr. Souza is a Research Fellow of the Coordenacao de Aperfeicoamento de Pessoal de Nível Superior at CAPES/Brazil.

Address correspondence to Dr. Weiss.