1887

Abstract

is an obligate intracellular bacterial pathogen that principally inhabits neutrophils. However, infection with results in a moderate to marked thrombocytopenia. In host neutrophils, uses sialylated ligands, primarily P-selectin glycoprotein ligand-1 (PSGL-1), to enter its host cell. PSGL-1 is expressed on a wide array of haematopoietic cells, including megakaryocytes. In this study, it was hypothesized that (i) cells of the megakaryocytic lineage (MEG-01 cells) would be susceptible to infection and (ii) infection may induce alterations in platelet production contributing to infection-induced thrombocytopenia. It was found that MEG-01 cells are susceptible to infection. MEG-01 cells expressing abundant sialylated ligands were the most susceptible to infection, and the absence of sialylation, or blocking of PSGL-1, limited infection susceptibility. However, infected MEG-01 cells produced proplatelets and platelet-like particles comparable to uninfected cells. These results highlight a novel target of pathogen infection and suggest that the pathogen may utilize similar strategies to gain access to megakaryocytes. Direct pathogen modification of platelet production may not play a role in infection-induced thrombocytopenia.

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2008-04-01
2019-10-18
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References

  1. Bakken, J. S., Krueth, J., Wilson-Nordskog, C., Tilden, R. L., Asanovich, K. & Dumler, J. S. ( 1996; ). Clinical and laboratory characteristics of human granulocytic ehrlichiosis. JAMA 275, 199–205.[CrossRef]
    [Google Scholar]
  2. Bakken, J. S., Aguero-Rosenfeld, M. E., Tilden, R. L., Wormser, G. P., Horowitz, H. W., Raffalli, J. T., Baluch, M., Riddell, D., Walls, J. J. & Dumler, J. S. ( 2001; ). Serial measurements of hematologic counts during the active phase of human granulocytic ehrlichiosis. Clin Infect Dis 32, 862–870.[CrossRef]
    [Google Scholar]
  3. Battinelli, E., Willoughby, S. R., Foxall, T., Valeri, C. R. & Loscalzo, J. ( 2001; ). Induction of platelet formation from megakaryocytoid cells by nitric oxide. Proc Natl Acad Sci U S A 98, 14458–14463.[CrossRef]
    [Google Scholar]
  4. Bedner, E., Burfeind, P., Hsieh, T. C., Wu, J. M., Aguero-Rosenfeld, M. E., Melamed, M. R., Horowitz, H. W., Wormser, G. P. & Darzynkiewicz, Z. ( 1998; ). Cell cycle effects and induction of apoptosis caused by infection of HL-60 cells with human granulocytic ehrlichiosis pathogen measured by flow and laser scanning cytometry. Cytometry 33, 47–55.[CrossRef]
    [Google Scholar]
  5. Bexfield, N. H., Villiers, E. J. & Herrtage, M. E. ( 2005; ). Immune-mediated haemolytic anaemia and thrombocytopenia associated with Anaplasma phagocytophilum in a dog. J Small Anim Pract 46, 543–548.[CrossRef]
    [Google Scholar]
  6. Borjesson, D. L., Simon, S. I., Tablin, F. & Barthold, S. W. ( 2001; ). Thrombocytopenia in a mouse model of human granulocytic ehrlichiosis. J Infect Dis 184, 1475–1479.[CrossRef]
    [Google Scholar]
  7. Borjesson, D. L., Simon, S. I., Hodzic, E., Ballantyne, C. M. & Barthold, S. W. ( 2002; ). Kinetics of CD11b/CD18 up-regulation during infection with the agent of human granulocytic ehrlichiosis in mice. Lab Invest 82, 303–311.[CrossRef]
    [Google Scholar]
  8. Borjesson, D. L., Brazzell, J. L. & Feferman, R. ( 2005a; ). Platelet dysfunction after association with Anaplasma phagocytophilum in vitro. Ann N Y Acad Sci 1063, 413–415.[CrossRef]
    [Google Scholar]
  9. Borjesson, D. L., Kobayashi, S. D., Whitney, A. R., Voyich, J. M., Argue, C. M. & Deleo, F. R. ( 2005b; ). Insights into pathogen immune evasion mechanisms: Anaplasma phagocytophilum fails to induce an apoptosis differentiation program in human neutrophils. J Immunol 174, 6364–6372.[CrossRef]
    [Google Scholar]
  10. Chen, S. M., Dumler, J. S., Bakken, J. S. & Walker, D. H. ( 1994; ). Identification of a granulocytotropic Ehrlichia species as the etiologic agent of human disease. J Clin Microbiol 32, 589–595.
    [Google Scholar]
  11. Chuang, J. L. & Schleef, R. R. ( 2001; ). Adenovirus-mediated expression and packaging of tissue-type plasminogen activator in megakaryocytic cells. Thromb Haemost 85, 1079–1085.
    [Google Scholar]
  12. Clarke, M. C., Savill, J., Jones, D. B., Noble, B. S. & Brown, S. B. ( 2003; ). Compartmentalized megakaryocyte death generates functional platelets committed to caspase-independent death. J Cell Biol 160, 577–587.[CrossRef]
    [Google Scholar]
  13. Dumler, J. S., Barbet, A. F., Bekker, C. P., Dasch, G. A., Palmer, G. H., Ray, S. C., Rikihisa, Y. & Rurangirwa, F. R. ( 2001; ). Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and ‘HGE agent’ as subjective synonyms of Ehrlichia phagocytophila. Int J Syst Evol Microbiol 51, 2145–2165.[CrossRef]
    [Google Scholar]
  14. Frenette, P. S., Denis, C. V., Weiss, L., Jurk, K., Subbarao, S., Kehrel, B., Hartwig, J. H., Vestweber, D. & Wagner, D. D. ( 2000; ). P-Selectin glycoprotein ligand 1 (PSGL-1) is expressed on platelets and can mediate platelet-endothelial interactions in vivo. J Exp Med 191, 1413–1422.[CrossRef]
    [Google Scholar]
  15. Ge, Y. & Rikihisa, Y. ( 2006; ). Anaplasma phagocytophilum delays spontaneous human neutrophil apoptosis by modulation of multiple apoptotic pathways. Cell Microbiol 8, 1406–1416.[CrossRef]
    [Google Scholar]
  16. Goodman, J. L., Nelson, C., Vitale, B., Madigan, J. E., Dumler, J. S., Kurtti, T. J. & Munderloh, U. G. ( 1996; ). Direct cultivation of the causative agent of human granulocytic ehrlichiosis. N Engl J Med 334, 209–215.[CrossRef]
    [Google Scholar]
  17. Goodman, J. L., Nelson, C. M., Klein, M. B., Hayes, S. F. & Weston, B. W. ( 1999; ). Leukocyte infection by the granulocytic ehrlichiosis agent is linked to expression of a selectin ligand. J Clin Invest 103, 407–412.[CrossRef]
    [Google Scholar]
  18. Herron, M. J., Nelson, C. M., Larson, J., Snapp, K. R., Kansas, G. S. & Goodman, J. L. ( 2000; ). Intracellular parasitism by the human granulocytic ehrlichiosis bacterium through the P-selectin ligand, PSGL-1. Science 288, 1653–1656.[CrossRef]
    [Google Scholar]
  19. Hodzic, E., Feng, S., Fish, D., Leutenegger, C. M., Freet, K. J. & Barthold, S. W. ( 2001; ). Infection of mice with the agent of human granulocytic ehrlichiosis after different routes of inoculation. J Infect Dis 183, 1781–1786.[CrossRef]
    [Google Scholar]
  20. Hsieh, T. C., Aguero-Rosenfeld, M. E., Wu, J. M., Ng, C., Papanikolaou, N. A., Varde, S. A., Schwartz, I., Pizzolo, J. G., Melamed, M. & other authors ( 1997; ). Cellular changes and induction of apoptosis in human promyelocytic HL-60 cells infected with the agent of human granulocytic ehrlichiosis (HGE). Biochem Biophys Res Commun 232, 298–303.[CrossRef]
    [Google Scholar]
  21. Italiano, J. E., Jr, Lecine, P., Shivdasani, R. A. & Hartwig, J. H. ( 1999; ). Blood platelets are assembled principally at the ends of proplatelet processes produced by differentiated megakaryocytes. J Cell Biol 147, 1299–1312.[CrossRef]
    [Google Scholar]
  22. Klein, M. B., Miller, J. S., Nelson, C. M. & Goodman, J. L. ( 1997; ). Primary bone marrow progenitors of both granulocytic and monocytic lineages are susceptible to infection with the agent of human granulocytic ehrlichiosis. J Infect Dis 176, 1405–1409.[CrossRef]
    [Google Scholar]
  23. Lester, S. J., Breitschwerdt, E. B., Collis, C. D. & Hegarty, B. C. ( 2005; ). Anaplasma phagocytophilum infection (granulocytic anaplasmosis) in a dog from Vancouver Island. Can Vet J 46, 825–827.
    [Google Scholar]
  24. Li, F., Erickson, H. P., James, J. A., Moore, K. L., Cummings, R. D. & McEver, R. P. ( 1996; ). Visualization of P-selectin glycoprotein ligand-1 as a highly extended molecule and mapping of protein epitopes for monoclonal antibodies. J Biol Chem 271, 6342–6348.[CrossRef]
    [Google Scholar]
  25. Madigan, J. E. & Gribble, D. ( 1987; ). Equine ehrlichiosis in northern California: 49 cases (1968–1981). J Am Vet Med Assoc 190, 445–448.
    [Google Scholar]
  26. Munderloh, U. G., Lynch, M. J., Herron, M. J., Palmer, A. T., Kurtti, T. J., Nelson, R. D. & Goodman, J. L. ( 2004; ). Infection of endothelial cells with Anaplasma marginale and A. phagocytophilum. Vet Microbiol 101, 53–64.[CrossRef]
    [Google Scholar]
  27. Ogura, M., Morishima, Y., Ohno, R., Kato, Y., Hirabayashi, N., Nagura, H. & Saito, H. ( 1985; ). Establishment of a novel human megakaryoblastic leukemia cell line, MEG-01, with positive Philadelphia chromosome. Blood 66, 1384–1392.
    [Google Scholar]
  28. Ogura, M., Morishima, Y., Okumura, M., Hotta, T., Takamoto, S., Ohno, R., Hirabayashi, N., Nagura, H. & Saito, H. ( 1988; ). Functional and morphological differentiation induction of a human megakaryoblastic leukemia cell line (MEG-01s) by phorbol diesters. Blood 72, 49–60.
    [Google Scholar]
  29. Pusterla, N., Pusterla, J. B., Braun, U. & Lutz, H. ( 1999; ). Experimental cross-infections with Ehrlichia phagocytophila and human granulocytic ehrlichia-like agent in cows and horses. Vet Rec 145, 311–314.[CrossRef]
    [Google Scholar]
  30. Reneer, D. V., Kearns, S. A., Yago, T., Sims, J., Cummings, R. D., McEver, R. P. & Carlyon, J. A. ( 2006; ). Characterization of a sialic acid- and P-selectin glycoprotein ligand-1-independent adhesin activity in the granulocytotropic bacterium Anaplasma phagocytophilum. Cell Microbiol 8, 1972–1984.[CrossRef]
    [Google Scholar]
  31. Sarkar, M., Reneer, D. V. & Carlyon, J. A. ( 2007; ). Sialyl-Lewis x-independent infection of human myeloid cells by Anaplasma phagocytophilum strains HZ and HGE1. Infect Immun 75, 5720–5725.[CrossRef]
    [Google Scholar]
  32. Scorpio, D. G., Akkoyunlu, M., Fikrig, E. & Dumler, J. S. ( 2004; ). CXCR2 blockade influences Anaplasma phagocytophilum propagation but not histopathology in the mouse model of human granulocytic anaplasmosis. Clin Diagn Lab Immunol 11, 963–968.
    [Google Scholar]
  33. Scorpio, D. G., Von Loewenich, F. D., Bogdan, C. & Dumler, J. S. ( 2005; ). Innate immune tissue injury and murine HGA: tissue injury in the murine model of granulocytic anaplasmosis relates to host innate immune response and not pathogen load. Ann N Y Acad Sci 1063, 425–428.[CrossRef]
    [Google Scholar]
  34. Takeuchi, K., Satoh, M., Kuno, H., Yoshida, T., Kondo, H. & Takeuchi, M. ( 1998; ). Platelet-like particle formation in the human megakaryoblastic leukaemia cell lines, MEG-01 and MEG-01s. Br J Haematol 100, 436–444.[CrossRef]
    [Google Scholar]
  35. Telford, S. R., III, Dawson, J. E., Katavolos, P., Warner, C. K., Kolbert, C. P. & Persing, D. H. ( 1996; ). Perpetuation of the agent of human granulocytic ehrlichiosis in a deer tick-rodent cycle. Proc Natl Acad Sci U S A 93, 6209–6214.[CrossRef]
    [Google Scholar]
  36. Wagers, A. J., Stoolman, L. M., Craig, R., Knibbs, R. N. & Kansas, G. S. ( 1998; ). An sLex-deficient variant of HL60 cells exhibits high levels of adhesion to vascular selectins: further evidence that HECA-452 and CSLEX1 monoclonal antibody epitopes are not essential for high avidity binding to vascular selectins. J Immunol 160, 5122–5129.
    [Google Scholar]
  37. White, J. G. ( 2005; ). Platelets are covercytes, not phagocytes: uptake of bacteria involves channels of the open canalicular system. Platelets 16, 121–131.[CrossRef]
    [Google Scholar]
  38. Wong, S. J. & Thomas, J. A. ( 1998; ). Cytoplasmic, nuclear, and platelet autoantibodies in human granulocytic ehrlichiosis patients. J Clin Microbiol 36, 1959–1963.
    [Google Scholar]
  39. Yago, T., Leppanen, A., Carlyon, J. A., Akkoyunlu, M., Karmakar, S., Fikrig, E., Cummings, R. D. & McEver, R. P. ( 2003; ). Structurally distinct requirements for binding of P-selectin glycoprotein ligand-1 and sialyl Lewis x to Anaplasma phagocytophilum and P-selectin. J Biol Chem 278, 37987–37997.[CrossRef]
    [Google Scholar]
  40. Yoshiie, K., Kim, H. Y., Mott, J. & Rikihisa, Y. ( 2000; ). Intracellular infection by the human granulocytic ehrlichiosis agent inhibits human neutrophil apoptosis. Infect Immun 68, 1125–1133.[CrossRef]
    [Google Scholar]
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