1887

Journal of Medical Microbiology logo

Volume 5, Issue 4

A Study Of Metabolic Factors Involved In The Intracellular Germination Of Spores After Phagocytosis Free

Abstract

SUMMARY

The environment within the metabolically active polymorphonuclear leucocyte during phagocytosis is conducive to the germination of phagocytically ingested spores of type A. Specific metabolic inhibitors of leucocyte function—glyceraldehyde, hydrocortisone, colchicine, methimazole and azide—were used to study metabolic pathways during phagocytosis that may be responsible for inducing spore germination within the leucocyte.

Leakage of Ca from labelled spores was taken as evidence of spore germination. Release of free Ca from phagocytosed spores in sporeleucocyte mixtures in in-vitro systems was normal in the presence of hydrocortisone, methimazole, azide and methimazole+azide. These findings indicate that HO production, iodination and myeloperoxidase activities during phagocytosis are not required for germination. In the presence of glyceraldehyde or colchicine, release of Ca from spore-leucocyte mixtures was profoundly inhibited and this indicates that glycolysis and lysosome degranulation are essential for intraleucocytic spore germination.

In leucocytes from patients with a genetic abnormality of phagocytosis, i.e., chronic granulomatous disease of childhood (CGD), germination of spores was significantly depressed. Since normal mechanisms of HO production and possibly of lysosomal degranulation are blocked in CGD leucocytes, the results suggest that HO and lysosomal enzymes are involved in triggering spore germination in this model. Nevertheless, a limited release of Ca was observed and this seems to indicate that small amounts of lysosomal enzymes are available for reaction with engulfed spores in CGD leucocytes.

The results of these studies with metabolic inhibitors and genetic blocks lead to the view that germination of spores of within leucocytes is triggered by the combined action of lowered H resulting from glycolysis together with lytic enzymes resulting from degranulation of lysosomes.

  • Received:
  • Accepted:
  • Published Online:
© J. MED. MICROBIOL. 1972
Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-5-4-381
1972-11-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/jmm/5/4/medmicro-5-4-381.html?itemId=/content/journal/jmm/10.1099/00222615-5-4-381&mimeType=html&fmt=ahah

References

  1. Baehner R. L., Karnovsky M. J., Karnovsky M. L. 1969; Degranulation of leukocytes in chronic granulomatous disease. J. Clin. Invest 48:187
     [Google Scholar]
  2. Booth R., Suzuki J. B., Grecz N. 1971a; Pathogenesis of Clostridium botulinum spores : in vivo fate ofC.botulinum typeAspores. Trans. Ill. State Acad. Sci 64:147
     [Google Scholar]
  3. Booth R., Suzuki J. B., Grecz N. 1971b; Sequential use of Wright’s and Ziehl-Neelsen’s stains in demonstrating phagocytosis of bacterial spores. Stain Technol 46:23
     [Google Scholar]
  4. Bona C., Macovschi O., Gheordunescu V. 1969; Dehydrogenase and oxidase activities of guinea pig leukocytes during pinocytosis. J. Reticuloendothel. Soc 6:271
     [Google Scholar]
  5. Bray G. A. 1960; A simple efficient liquid scintillator for counting aqueous solution in a liquid scintillation counter. Analyt. Biochem 1:279
     [Google Scholar]
  6. Cohn Z. A. 1968; The structure and function of monocytes and macrophages. Adv. Immun 7:163
     [Google Scholar]
  7. Good R. A., Quie P. G., Windhorst D. B., Page A. R., Rodey G. E., White J., Wolfson J. J., Holmes B.H. 1968; Fatal (chronic) granulomatous disease of childhood: a hereditary defect of leukocyte function. Semin. Hematol 5:215
     [Google Scholar]
  8. Gould G. W., Hitchins A. D. 1963; Sensitization of bacterial spores to lysozyme and to hydrogen peroxide with agents which rupture disulphide bonds. J. Gen. Microbiol 33:413
     [Google Scholar]
  9. Grecz N., Anellis A., Schneider M. D. 1962; Procedure for cleaning of Clostridium botulinum spores. J. Bact 84:552
     [Google Scholar]
  10. Holmes B., Page A. R., Good R. A. 1967; Studies of the metabolic activity of leukocytes from patients with a genetic abnormality of phagocytic function. J. Clin. Invest 46:1422
     [Google Scholar]
  11. Kauder E., Kahle L. L., Moreno H., Partin J. C. 1968; Leukocyte degranulation and vacuole formation in patients with chronic granulomatous disease of childhood. J. Clin. Invest 47:1753
     [Google Scholar]
  12. Klebanoff S. J. 1968; Myeloperoxidase-halide-hydrogen peroxide antibacterial system. J. Bact 95:2131
     [Google Scholar]
  13. Klebanoff S. J. 1970; Myeloperoxidase : contribution to the microbicidal activity of intact leukocytes. Science N. Y. 169:1095
     [Google Scholar]
  14. Klebanoff S. J., White L. R. 1969; Iodination defect in the leukocytes of a patient with chronic granulomatous disease of childhood. New Engl. J. Med 280:460
     [Google Scholar]
  15. Malawista S. E., Bodel Phyllis T. 1967; The dissociation by colchicine of phagocytosis from increased oxygen consumption in human leukocytes. J. Clin. Invest 46:786
     [Google Scholar]
  16. Mandell G. L., Rubin W., Hook E. W. 1970; The effect of an NADH oxidase inhibitor (hydrocortisone) on polymorphonuclear leukocyte bactericidal activity. J. Clin. Invest 49:1381
     [Google Scholar]
  17. Morton D. J., Moran J. F., Stjernholm R. L. 1969; Carbohydrate metabolism in leukocytes. XI. Stimulation of eosinophils and neutrophils. J. Reticuloendothel. Soc 6:525
     [Google Scholar]
  18. Murrell W. G. 1961; Spore formation and germination as a microbial reaction to the environment. Symp. Soc. Gen. Microbiol 11:100
     [Google Scholar]
  19. Paul B. B., Strauss R. R., Jacobs A. A., Sbarra A.J. 1970; Function of H2O2, myeloperoxidase, and hexose monophosphate shunt enzymes in phagocytizing cells from different species. Infect. Immun 1:338
     [Google Scholar]
  20. Powell Joan F., Strange R. E. 1953; Biochemical changes occurring during the germination of bacterial spores. Biochem. J 54:205
     [Google Scholar]
  21. Quie P. G., White J. G., Holmes B., Good R. A. 1967; In vitro bactericidal capacity of human polymorphonuclear leukocytes: diminished activity in chronic granulomatous disease of childhood. J. Clin. Invest 46:668
     [Google Scholar]
  22. Rous P. 1925; The relative reaction within living mammalian tissues. II. On the mobilization of acid material within cells, and the reaction as influenced by the cell state. J. Exp. Med 41:399
     [Google Scholar]
  23. Rowley D. 1962; Phagocytosis. Adv. Immun 2:241
     [Google Scholar]
  24. Selvaraj R., J Sbarra A. J. 1966; Relationship of glycolytic and oxidative metabolism to particle entry and destruction in phagocytosing cells. Nature, Lond 211:1272
     [Google Scholar]
  25. Suzuki J. B., Booth R., Grecz N. 1970; Pathogenesis of Clostridium botulinum type A: release of toxin from C. botulinum spores in vitro by leukocytes. Res. Commun. Chem. Path. Pharmac 1:691
     [Google Scholar]
  26. Suzuki J. B., Booth R. R., Grecz N. 1971a; Evaluation of phagocytic activity by ingestion of labelled bacteria. J. Infect. Dis 123:93
     [Google Scholar]
  27. Suzuki J. B., Booth R. R., Grecz N. 1971b; In-vivo and in-vitro release of Ca45 from spores of Clostridium botulinum type A as further evidence for spore germination. Res. Commun. Chem. Path. Pharmac 2:16
     [Google Scholar]
  28. Suzuki J. B., Booth R., Benedik A., Grecz N. 1971; Pathogenesis of Clostridium botulinum type A: study of in vivo toxin release by implantation of diffusion chambers containing spores, vegetative cells and free toxin. Infect. Immun 3:659
     [Google Scholar]
  29. Wynne E. S. 1957; Symposium on bacterial spore germination. Bact. Rev 21:259
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-5-4-381
Loading
A Study Of Metabolic Factors Involved In The Intracellular Germination Of Clostridium Botulinum Spores After Phagocytosis
J. Med. Microbiol. 5, 381 (1972); https://doi.org/10.1099/00222615-5-4-381
/content/journal/jmm/10.1099/00222615-5-4-381
/content/journal/jmm/10.1099/00222615-5-4-381
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error