1887

Abstract

SUMMARY

The destruction of secondary mouse-embryo (ME) cells by was studied by indirect immunofluorescence with ME-cell antiserum as a specific label to trace the fate of mammalian-cell cytoplasm. The appearance of naegleria-induced cytopathic effect in the cultures coincided with the accumulation of discrete particles containing granules of ME-cell antigen within the cytoplasm of amoebae, suggesting that the organisms ingested host-cell material. In cultures containing cytochalasin B, a non-lethal inhibitor of phagocytosis by trophozoites failed to acquire any granular fluorescence and were not cytopathogenic. The engulfment of mammalian-cell cytoplasm by the organisms was confirmed when thin sections of naegleria-infected ME-cell cultures were examined by electron microscopy. Amoebae were seen in the process of detaching portions of cytoplasm from whole ME cells by means of distinctive ingesting pseudopodia, and fragments of mammalian-cell cytoplasm were identified within the food vacuoles of trophozoites. There was no evidence for cytotoxic disruption of ME cells before or during engulfment of these fragments. It is concluded that trophozoites attack and destroy cultured ME cells by a phagocytosis-like mechanism alone, without the aid of any amoeba-associated cytotoxic or cytolytic agents. The possible significance of these findings with respect to the in-vivo pathocity of is discussed.

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1979-08-01
2024-03-28
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References

  1. Allison A. C., Davies P., de Petris S. 1971; Role of contractile microfilaments in macrophage movement and endocytosis. Nature, New Biol 232:153
    [Google Scholar]
  2. Axline S. G., Reaven E. 1972; In vitro inhibition of phagocytosis and disruption of microfilaments by cytochalasin B. J. clin. Invest 51:6a
    [Google Scholar]
  3. Bessis M. 1970; Cell death. Triangle 9:191
    [Google Scholar]
  4. Brown T. 1978; Observations by light microscopy on the cytopathogenicity of Naegleria fowled in mouse embryo-cell cultures. J. med. Microbiol 11:249
    [Google Scholar]
  5. Brown T. 1979; Inhibition by amoeba-specific antiserum and by cytochalasin B of the cyto pathogenicity of Naegleria fowleri in mouse embryo-cell cultures. J. med. Microbiol 12:353
    [Google Scholar]
  6. Butt C. G., Baro C., Knorr R. W. 1968; Naegleria (sp.) identified in amebic encephalitis. Am. J. clin. Path 50:568
    [Google Scholar]
  7. Carter R. F. 1972; Primary amoebic meningo-encephalitis. An appraisal of present knowledge. Trans. R. Soc. trop. Med. Hyg 66:193 (see discussion p. 209)
    [Google Scholar]
  8. Chang S. L. 1971; Small, free-living amebas: cultivation, quantitation, identification, classification, pathogenesis, and resistance. Curr. Top. comp. Pathobiol 1:201
    [Google Scholar]
  9. Chang S. L. 1974; Etiological, pathological, epidemiological and diagnostical considerations of primary amoebic meningoencephalitis. C.R.C. crit. Rev. Microbiol 3:135
    [Google Scholar]
  10. Culbertson C. G. 1971; The pathogenicity of soil amebas. A. Rev. Microbiol 25:231
    [Google Scholar]
  11. Cursons R. T. M., Brown T. J. 1978; Use of cell cultures as an indicator of pathogenicity of free-living amoebae. J. clin. Path 31:1
    [Google Scholar]
  12. Cursons R. T. M., Brown T. J., Keys E. A. 1978; Virulence of pathogenic free-living amoebae. J. Parasit 64:744
    [Google Scholar]
  13. Duma R. J., Ferrell H. W., Nelson E. C., Jones M. M. 1969; Primary amebic meningoencephalitis. New Engl. J. Med 281:1315
    [Google Scholar]
  14. Duma R. J., Rosenblum W. I., McGehee R. F., Jones M. M., Nelson E. C. 1971; Primary amoebic meningoencephalitis caused by Naegleria. Two new cases, response to amphotericin B, and a review. Ann. intern.Med 74:923
    [Google Scholar]
  15. Eaton R. D. P., Meerovitch E., Costerton J. W. 1969; A surface-active lysosome in Entamoeba histolytica. Trans. R. Soc. trop. Med. Hyg 63:678
    [Google Scholar]
  16. Eaton R. D. P., Meerovitch E., Costerton J. W. 1970; The functional morphology of pathogenicity in Entamoeba histolytica. Ann. trop. Med. Parasit 64:299
    [Google Scholar]
  17. Fulton C. 1970; Amebo-flagellates as research partners: the laboratory biology of naegleria and tetramitus. In Methods in cell physiology, 4 edited by Prescott D. M. New York and London: p 341
    [Google Scholar]
  18. Griffin J. L. 1972; Human amebic dysentery. Electron microscopy of Entamoeba histolytica contacting, ingesting and digesting inflammatory cells. Am. J. trop. Med. Hyg 21:895
    [Google Scholar]
  19. Griffin J. L., Juniper K. 1971; Ultrastructure of Entamoeba histolytica from human amebic dysentery. Archs. Path 91:271
    [Google Scholar]
  20. Hartwig J. H., Stossel T. P. 1976; Interactions of actin, myosin, and an actin-binding protein of rabbit pulmonary macrophages. III. Effects of cytochalasin B. J. cell Biol 71:295
    [Google Scholar]
  21. Jarumilinta R., Kradolfer F. 1964; The toxic effect of Entamoeba histolytica on leucocytes. Ann. trop. Med. Parasit 58:375
    [Google Scholar]
  22. Lastovica A. J. 1976; Microfilaments in Naegleria fowleri amoebae. Z. ParasitKde 50:245
    [Google Scholar]
  23. Lastovica A. J., Dingle A. D. 1971; Superprecipitation of an actomyosin-like complex isolated from Naegleria gruberi amoebae. Expl cell Res 66:337
    [Google Scholar]
  24. Maitra S. C., Krishna Prasad B. N., Das S. R., Agarwala S. C. 1974; Study of Naegleria aerobia by electron microscopy. Trans. R. Soc. trop. Med. Hyg 68:56
    [Google Scholar]
  25. Maitra S. C., Krishna Prasad B. N., Agarwala S. C., Das S. R. 1976; Ultrastructural studies on experimental primary amoebic meningoencephalitis (PAME) of mouse due to Naegleria aerobia and Hartmannella culbertsoni. Int. J. Parasit 6:489
    [Google Scholar]
  26. Martinez A. J., Duma R. J., Nelson E. C., Moretta F. L. 1973; Experimental naegleria meningoencephalitis in mice. Penetration of the olfactory mucosal epithelium by naegleria and pathologic changes produced: a light and electron microscope study. Lab. Invest 29:121
    [Google Scholar]
  27. Martinez A. J., Nelson E. C., Jones M. M., Duma R. J., Rosenblum W. I. 1971; Experimental naegleria meningoencephalitis in mice. An electron microscope study. Lab. Invest 25:465
    [Google Scholar]
  28. Neal R. A. 1971; Progress report. Pathogenesis of amoebiasis. Gut 12:483
    [Google Scholar]
  29. Newton B. A. 1972; Protozoal pathogenicity. In Microbial pathogenicity in man and animals Symposium no. 22 of the Society for General Microbiology, edited by Smith H., Pearce J. H. Cambridge: p 269
    [Google Scholar]
  30. Persijn J. P., Scherft J. P. 1965; Sheet mica—a nonadherent carrier for surface culture of cells to be embedded in Epon. Stain technol 40:89
    [Google Scholar]
  31. Plagemann P. G. W., Graff J. C., Wohlhueter R. M. 1977; Binding of [3H] cytochalasin B and its relationship to inhibition of hexose transport in Novikoff rat hepatoma cells. J. biol. Chem 252:4191
    [Google Scholar]
  32. Pollard T. D., Korn E. D. 1973a; Acanthamoeba myosin. I. Isolation from Acanthamoeba castellanii of an enzyme similar to muscle myosin. J. biol. Chem 248:4682
    [Google Scholar]
  33. Pollard T. D., Korn E. D. 1973b; Acanthamoeba myosin. II. Interaction with actin and with a new cofactor protein required for actin activation of Mg2 + adenosine triphosphatase activity. J. biol. Chem 248:4691
    [Google Scholar]
  34. Pollard T. D., Stafford W. F., Porter M. E. 1978; Characterization of a second myosin from Acanthamoeba castellanii. J. biol. Chem 253:4798
    [Google Scholar]
  35. Pollard T. D., Weihing R. R. 1974; Actin and myosin and cell movement. C.R.C. crit. Rev. Biochem 2:1
    [Google Scholar]
  36. Reaven E. P., Axline S. G. 1973; Subplasmalemmal microfilaments and microtubules in resting and phagocytizing cultivated macrophages. J. cell. Biol 59:12
    [Google Scholar]
  37. Reynolds E. S. 1963; The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. cell Biol 17:208
    [Google Scholar]
  38. Schuster F. 1963; An electron microscope study of the amoebofiagellate, Naegleria gruberi (Schardinger). I. The amoeboid and flagellate stages. J. Protozool 10:297
    [Google Scholar]
  39. Smith H. 1972; The little-known determinants of microbial pathogenicity. In Microbial pathogenicity in man and animals Symposium no. 22 of the Society for General Microbiology, edited by Smith H., Pearce J. H. Cambridge: p 1
    [Google Scholar]
  40. Stossel T. P. 1974a; Phagocytosis; I. New Engl. J. Med 290:717
    [Google Scholar]
  41. Stossel T. P. 1974b; Phagocytosis; II. New Engl. J. Med 290:774
    [Google Scholar]
  42. Visvesvara G. S., Callaway C. S. 1974; Light and electron microscopic observations on the pathogenesis of Naegleria fowleri in mouse brain and tissue culture. J. Protozool 21:239
    [Google Scholar]
  43. Wagner P. 1978; What is a myosin?: Acanthamoeba revisited. Nature, Lond 274:846
    [Google Scholar]
  44. Weihing R. R., Korn E. D. 1971; Acanthamoeba actin. Isolation and properties. Biochemistry, N.Y 10:590
    [Google Scholar]
  45. Wessells N. K., Spooner B. S., Ash J. F., Bradley M. O., Luduena M. A., Taylor E. L., Wrenn J. T., Yamada K. M. 1971; Microfilaments in cellular and developmental processes. Science, N.Y 171:135
    [Google Scholar]
  46. Westphal A., Michel R., Muhlpfordt H. 1972; Elektronenmikroskopische Befunde bei der Phagozytose von Crithidia sp. durch Entamoeba histolytica. Z. Tropenmed. Parasit 23:117
    [Google Scholar]
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