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Parasitology Research

, Volume 77, Issue 1, pp 59–64 | Cite as

Monoclonal antibodies identify micronemes and a new population of cytoplasmic granules cross-reacting with micronemes of cystozoites ofSarcocystis muris

  • R. Entzeroth
  • A. König
  • J. -F. Dubremetz
Original Investigations

Abstract

Micronemes of cystozoites ofSarcocystis muris were isolated after subcellular fractionation and used for immunization of BALB/C mice. After spleen cells of immunized mice were fused with SP20 myeloma cells, ten different monoclonal antibodies (mAbs) were isolated. These antibodies reacted with antigens whose molecular weight ranged from 16 to >90 kDa. Six mAbs recognized granules of 150–400 nm that were located in the vicinity of the Golgi complex but were not identical with dense granules. Two mAbs (2A3, 3A8) were specific for micronemes of cystozoites as demonstrated by immunoelectron microscopy. However, these antibodies also recognized the population of granules near the Golgi complex. Cross-reactivity between micronemes and a dense granule population has not previously been reported. Host cells that had been contacted by cystozoites showed patchy fluorescence when probed with mAb 2A3. This suggests that microneme antigens could be transferred to the host-cell surface during parasitehost cell interactions.

Keywords

Molecular Weight Monoclonal Antibody Host Cell Myeloma Spleen Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Bannister LH, Mitchell GH (1989) The fine structure of secretion byPlasmodium knowlesi merozoites during red cell invasion. J Protozool 36:362–367Google Scholar
  2. Brown GV, Culvenor JG, Crewttier PE, Bianco AE, Coppel RL, Saint RB, Stahl MD, Kemp DJ, Anders RF (1985) Localization of the ring-infected erythrocyte surface antigen (RESA) ofPlasmodium falciparum in merozoites and ring-infected erythrocytes. J Exp Med 162:774–779Google Scholar
  3. Dubremetz JF, Dissous C (1980) Characteristic proteins of micronemes and dense granules fromSarcocystis tenella zoites (Protozoa, Coccidia). Mol Biochem Parasitol 1:279–289Google Scholar
  4. Dubremetz JF, Ferreira E, Dissous C (1989) Isolation and characterization of rhoptries and micronemes fromEimeria nieschulzi zoites (Sporozoa, Coccidia). Parasitol Res 75:449–455Google Scholar
  5. Entzeroth R (1985) Invasion and early development ofSarcocystis muris (Apicomplexa, Sarcocystidae) in tissue cultures. J Protozool 32:446–453Google Scholar
  6. Entzeroth R, Dubremetz JF, Hodick D, Ferreira E (1986) Immunoelectron microscopic demonstration of the exocytosis of dense granule contents into the secondary parasitophorous vacuole ofSarcocystis muris (Protozoa, Apicomplexa). Eur J Cell Biol 41:182–188Google Scholar
  7. Galfre G, Howe SC, Milstein C, Butcher GW, Howard JC (1977) Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature 266:550–552Google Scholar
  8. Kimata I, Tanabe K (1987) Secretion byToxoplasma gondii of an antigen that appears to become associated with the parasitophorous vacuole membrane upon invasion of the host cell. J Cell Sci 88:231–239Google Scholar
  9. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  10. Nichols BA, Chiappino ML, O'Connor GR (1983) Secretion from the rhoptries ofToxoplasma gondii during host-cell invasion. J Ultrastruet Res 83:85–98Google Scholar
  11. Pohl U, Dubremetz JF, Entzeroth R (1989) Characterization and immunolocalization of the protein contents of micronemes ofSarcocystis muris cystozoites (Protozoa, Apicomplexa). Parasitol Res 75:199–205Google Scholar
  12. Quakyl IA, Matsumoto Y, Carter R, Udomsangoetch, R, Sjolander A, Berzins K, Perlmann P, Aikawa M, Miller LH (1989) Movement of a falciparum malaria protein through the erythrocyte cytoplasm to the erythrocyte membrane is associated with lysis of the erythrocyte and release of gametes. Infect Immun 57:833–839Google Scholar
  13. Roth JM, Bendayan E, Carlemalm W, Villinger M, Garavito M (1981) Enhancement of structural preservation and immunocytochemical staining in low-temperature embedded pancreatic tissue. J Histochem Cytochem 29:663–671Google Scholar
  14. Sam-Yellowe TY, Shio H, Perkins ME (1988) Secretion ofPlasmodium falciparum rhoptry protein into the plasma membrane of host erythrocytes. J Cell Biol 106:1507–1513Google Scholar
  15. Scholtyseck E, Mehlhorn H (1970) Ultrastructural study of characteristic organelles (paired organelles, micronemes, micropores) of Sporozoa and related organisms. Z Parasitenkd 34:97–127Google Scholar
  16. Slot JM, Geuze HJ (1985) A new method of preparing gold probes for multiple-labelling cytochemistry. Eur J Cell Biol 38:87–93Google Scholar
  17. Towbin H, Staehelin T, Gordon J (1978) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc natl Acad Sci USA 76:4350–4354Google Scholar
  18. Torii M, Matsumoto Y, Kamboj KK, Maracic M, Guo SQ (1989) Association of microneme antigens ofPlasmodium brasilianum merozoites with knobs and other parasite-induced structures in host erythrocytes. Infect Immun 57:596–601Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • R. Entzeroth
    • 1
  • A. König
    • 1
  • J. -F. Dubremetz
    • 2
  1. 1.Zoologisches InstitutBonn 1Federal Republic of Germany
  2. 2.U 42INSERMVilleneuve-D'AscqFrance

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