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

Abstract

T H E chemical environment within the metabolically active leucocyte during the phagocytic act is conducive to intracellular spore germination. This leucocyte-induced germination of Clostridium botulinum type A spores has been studied in various in-vitro systems after Booth, Suzuki and Grecz (1971a) reported that heat-resistant spores of this organism are converted to heatsensitive germinated spores and vegetative cells in the peritoneal cavity after they have been injected intraperitoneally (IP) into mice.These workers used new staining techniques (Booth, Suzuki and Grecz, 19716) to demonstrate phagocytosis by leucocytes in vivo and in vitro and to show subsequent germination of the spores within the leucocyte. Furthermore, Suzuki, Booth and Grecz (1970) recently found that Cl. botulinum spores must be actively phagocytosed to release toxin. The production of an equivalent ionic environment or exposure of the spores to leucocyte cytoplasmic extracts did not, by themselves, induce spore germination or toxin release. Studies involving the implantation of Millipore diffusion chambers containing spores alone or spores plus leucocytes into the peritoneal cavity of guinea-pigs further supported this concept (Suzuki, Booth, Benedik and Grecz, 1971). Ten per cent. of the animals given chambers containing spores alone died with typical symptoms of botulism, whereas chambers containing spores plus leucocytes killed 90 per cent. of the test guinea-pigs. Passively immunised guinea-pigs were able to survive the implantation of chambers containing C Z . botulinum spores plus leucocytes. Thus, it has been established that polymorphonuclear (PMN) leucocytes can provoke a pathogenic state when interacting with Cl. botulinum spores.During phagocytic engulfment, lactic acid accumulates as a result of glycolysis and causes a rapid decrease in pH of the phagocytic vacuole (Cohn, 1968) to values as low aspH 3.0-4-5 (Rous, 1925;Rowley, 1962). The pentose phosphate pathway present in peripheral leucocytes (Bona, Macovschi and Gheordunescu, 1969 ;Morton, Moran and Stjernholm, 1969) generates its metabolic by-product H202 and brings the reaction of the phagocytic vacuole to near neutrality. Concurrently, lysosome degranulation proceeds and brings into the proximity of the phagocytic vacuole a multitude of enzymes that include lysozyme and myeloperoxidase; the latter forms an effective bactericidal complex with H202 (Klebanoff, 1968(Klebanoff, , 1970Paul et al., 1970).The purpose of the present study was to determine the relationship of metabolic processes within the PMN leucocyte to the induction of intracellular CZ. botulinum spore germination and subsequent toxin release.

Immunologic Profile of Juvenile Periodontitis: II. Neutrophil Chemotaxis, Phagocytosis and Spore Germination

“…Glycolysis and the pentose phosphate pathway are particularly involved in triggering the germination of Clostridium botulinum spores in pathogenesis ofwound infections. 30 Phagocytic leukocytes from male patients with chronic granulomatous disease of childhood are greatly reduced in their capacity to induce intracellular bacterial spore germination compared to healthy controls.…”