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Axenic cultivation and characterization of Leishmania mexicana amastigote-like forms

  • P. A. Bates (a1), C. D. Robertson (a1), L. Tetley (a2) and G. H. Coombs (a1)

A new method is described which has made possible the long-term axenic cultivation of Leishmania mexicana amastigotelike forms in Schneider's Drosophila medium supplemented with 2% (v/v) foetal calf serum. Unlike previous methods, it utilizes direct culture of parasites obtained from the lesions of infected animals rather than adaptation of promastigotes in vitro. Ultrastructural (possession of megasomes), biochemical (cysteine proteinase activity and gelatin SDS-PAGE banding pattern) and infectivity (in vivo) data are presented which show the close similarity of the cultured forms to lesion amastigotes. The axenically cultured forms grew optimally at a temperature of 32–33 °C, providing further evidence for their amastigote nature. It was found that adjustment of the pH of the growth medium to 5·4 was required in order to retain the amastigote morphology of the cultured parasites. This supports the notion that leishmanial amastigotes are acidophiles.

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J. Alexander & K. Vickerman (1975). Fusion of host cell secondary lysosomes with the parasitophorous vacuoles of Leishmania mexicana-infected macrophages. Journal of Protozoology 22, 502–8.

R. L. Berens & J. J. Marr (1979). Growth of Leishmania donovani amastigotes in a continuous macrophage-like cell culture. Journal of Protozoology 26, 453–6.

R. L. Berens , R. Brun & S. M. Krassner (1976). A simple monophasic medium for axenic culture of hemoflagellates. Journal of Parasitology 62, 360–5.

D. Biegel , G. Topper & M. Rabinovitch (1983). Leishmania mexicana: temperature sensitivity of isolated amastigotes and of amastigotes infecting macrophages in culture. Experimental Parasitology 56, 289–97.

R. P. Brazil (1978). Isolation of the intracellular stages of Leishmania mexicana amazonensis using cellulose column. Annals of Tropical Medicine and Parasitology 72, 579–80.

K.-P. Chang (1980 a). Human cutaneous leishmania in a mouse macrophage cell line: propagation and isolation of intracellular parasites. Science 209, 1240–2.

K.-P. Chang & D. M. Dwyer (1976). Multiplication of a human parasite (Leishmania donovani) in phagolysosomes of hamster macrophages in vitro. Science 193, 678–80.

K.-P. Chang & D. M. Dwyer (1978). Leishmania donovani. Hamster macrophage interactions in vitro: cell entry, intracellular survival, and multiplication of amastigotes. Journal of Experimental Medicine 147, 515–30.

G. E. Childs , M. J. McRoberts & K. A. Foster (1976). Partial purification of amastigotes from cutaneous lesions of American leishmaniasis. Journal of Parasitology 62, 676–9.

P. S. Doyle , J. C. Engel , P. F. Pimenta , P. P. Da Silva & D. M. Dwyer (1991). Leishmania donovani: long-term culture of axenic amastigotes at 37 °C. Experimental Parasitology 73, 326–34.

S. Eperon & D. McMahon-Pratt (1989 b). Extracellular amastigote-like forms of Leishmania panamensis and L. braziliensis. II. Stage- and species-specific monoclonal antibodies. Journal of Protozoology 36, 510–18.

T. A. Glaser , J. E. Baatz , G. P. Kreishman & A. J. Mukkada (1988). pH homeostasis in Leishmania donovani amastigotes and promastigotes. Proceedings of the National Academy of Sciences, USA85, 7602–6.

T. A. Glaser , S. J. Wells , T. W. Spithill , J. M. Pettitt , D. C. Humphris & A. J. Mukkada (1990). Leishmania major and L. donovani: a method for rapid purification of amastigotes. Experimental Parasitology 71, 343–5.

B. C. Lockwood , M. J. North , D. J. Mallinson & G. H. Coombs (1987). Analysis of Leishmania proteinases reveals developmental changes in species- specific forms and a common 68-kDa activity. FEMS Microbiology Letters 48, 345–50.

D. L. Looker , S. Martinez , J. M. Horton & J. J. Marr (1986). Growth of Leishmania donovani amastigotes in the continuous human macrophage cell line U937: studies of drug efficacy and metabolism. Journal of Infectious Diseases 154, 323–7.

J. C. Meade , T. A. Claser , P. F. Bonventre & A. J. Mukkada (1984). Enzymes of carbohydrate metabolism in Leishmania donovani amastigotes. Journal of Protozoology 31, 156–61.

A. J. Mukkada , J. C. Meade , T. A. Glaser & P. F. Bonventre (1985). Enhanced metabolism of Leishmania donovani amastigotes at acid pH: an adaptation for intracellular growth. Science 229, 1099–101.

M. J. North & G. H. Coombs (1981). Proteinases of Leishmania mexicana amastigotes and promastigotes: analysis by gel electrophoresis. Molecular and Biochemical Parasitology 3, 293300.

A. A. Pan (1984). Leishmania mexicana: Serial cultivation of intracellular stages in a cell-free medium. Experimental Parasitology 58, 7280.

A. A. Pan & B. M. Honigberg (1985). Leishmania mexicana pifanoi: in vivo and in vitro interactions between amastigotes and macrophages. Zeitschrift für Parasitenkunde 71, 313.

A. A. Pan , D. McMahon-Pratt & B. M. Honigberg (1984). Leishmania mexicana pifanoi: antigenic characterization of promastigote and amastigote stages by solid-phase radioimmunoassay. Journal of Parasitology 70, 834–5.

A. A. Pan & S. C. Pan (1986). Leishmania mexicana: comparative fine structure of amastigotes and promastigotes in vitro and in vivo. Experimental Parasitology 62, 254–65.

M. F. Pupkis , L. Tetley & G. H. Coombs (1986). Leishmania mexicana: amastigote hydrolases in unusual lysosomes. Experimental Parasitology 62, 2939.

P. M. Rainey , T. W. Spithill , D. McMahon-Pratt & A. A. Pan (1991). Biochemical and molecular characterization of Leishmania pifanoi amastigotes in continuous axenic culture. Molecular and Biochemical Parasitology 49, 111–18.

C. D. Robertson & G. H. Coombs (1990). Characterisation of three groups of cysteine proteinases in the amastigotes of Leishmania mexicana mexicana. Molecular and Biochemical Parasitology 42, 269–76.

E. M. B. Saraiva , P. F. P. Pimenta , M. E. A. Pereira & W. De Souza (1983). Isolation and purification of amastigotes of Leishmania mexicana amazonensis by a gradient of metrizamide. Journal of Parasitology 69, 627–9.

P. A. Scott , S. James & A. Sher (1985). The respiratory burst is not required for killing of intracellular and extracellular parasites by a lymphokine-activated macrophage cell line. European Journal of Immunology, 15, 553–8.

R. M. Smejkal , R. Wolff & J. G. Olenick (1988). Leishmania braziliensis panamensis: increased infectivity resulting from heat shock. Experimental Parasitology 65, 19.

D. Zilberstein , N. Blumenfeld , V. Liveanu , A. Gepstein & C. L. Jaffe (1991). Growth at acidic pH induces an amastigote stage-specific protein in Leishmania promastigotes. Molecular and Riochemical Parasitology 45, 175–8.

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