Evaluation of Phagocytic Activity by Ingestion of Labeled Bacteria

Suzuki, Jon B.; Booth, Ron; Grecz, N.

doi:10.1093/infdis/123.1.93

Cited by 15 publications

(15 citation statements)

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“…The results of the present study concur with previous findings that spores of Clostridium botulinum are capable of germination after phagocytosis by polymorphonuclear (PMN) leucocytes (Suzuki, Booth and Grecz, 1970 Suzuki and Grecz, 1971a). The inhibitors used in the present study did not prevent spore germination per se.…”

Section: Discussionsupporting

confidence: 92%

“…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.…”

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

Suzuki

Grecz

1972

Journal of Medical Microbiology

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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.

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Section: Discussionsupporting

confidence: 92%

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

Suzuki

Grecz

1972

Journal of Medical Microbiology

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“…However, many of the studies in the past have been confined to the phagocytosis and minor attention has been paid to the fate of intracellular bacteria (for review see 1, 2 ) . A major problem has remained the elimination of nonphagocytized bacteria in an in vitro phagocytic system in order to study the dynamics of the intracellular phase (6,7,11). By our method, extracellular bacteria are effectively eliminated by antibiotics which cause no inactivation of phagocytized bacteria (8).…”

Section: Discussionmentioning

confidence: 99%

Influence of Phenylbutazone on the Phagocytic and Bactericidal Activities of Neutrophil Granulocytes

Solberg

1974

Acta Pathologica Microbiologica Scandinavica Section B Microbio

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The influence of phenylbutazone on the phagocytosis and intracellular killing of bacteria by human neutrophil granulocytes has been examined using a method which facilitates a precise in vitro evaluation of the phagocytic and bactericidal activities of polymorphonuclear leucocytes. In high concentrations, phenylbutazone caused a marked reduction in phagocytic activity and abruptly stopped the intracellular killing of bacteria by the granulocytes. No permanent effect of the drug upon resting cells was observed.

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“…Neutrophil phagocytosis was assessed using 45Ca-labeled cereus spores ( 10:1 ratio of spores to cells) by methods previously described. 27 Briefly, after incubation of 1 x 106/ml neutrophils and 1 x 107/ml 45Ca-labeled bacteria spores for 15 minutes in a shaking water bath at 37°C, filter-associated bacteria labeled with 45Ca were separated from free bacteria by filtration through a 5-µ Millipore filter. ** The counts per minute (CPM) associated with the filter were determined using the P-32 channel of a liquid scintillation counter (Beckman LS 3140T).…”

Section: Methodsmentioning

confidence: 99%

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

et al. 1984

Journal of Periodontology

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Patients with juvenile periodontitis (JP) were grouped into one of the two recognized forms of the disease: a localized form affecting incisors and first molars (LJP) and a generalized form affecting more than 14 teeth (GJP). The role of the neutrophil in the etiology and pathogenesis of LJP has recently been recognized. Experiments aimed at confirming previous information related to neutrophil chemotaxis defects in LJP were performed. Additional experiments aimed at demonstrating significant differences in phagocytosis of bacterial spores and inducing spore germination were also completed. Age- and sex-matched healthy subjects were used as "internal" controls (experiments run concomitantly). Peripheral blood neutrophils from 29 LJP, 24 GJP and 24 healthy subjects were assessed for chemotaxis. This assay was performed in a Boyden diffusion chamber using 10(-8) M N-formylmethionylleucylphenylalanine as chemoattractant. A chemotactic defect was defined being 2 standard deviations below the mean of healthy subjects. Twenty-three of 29 LJP (79%) and 14 of 25 GJP (58%) had a neutrophil defect. Peripheral blood neutrophil phagocytosis assays were performed in vitro using radiolabeled bacterial spores [45Ca]Bacillus cereus. Eighteen of 29 LJP (62%) and 7 of 24 GJP (29%) had a phagocytic defect. Neutrophil-induced spore germination assessed in vitro showed that 13 of 20 LJP (65%) and 3 of 8 GJP (38%) had this defect. These observations indicate that patients with JP have abnormalities in neutrophil functions of chemotaxis, phagocytosis and spore germination.

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Evaluation of Phagocytic Activity by Ingestion of Labeled Bacteria

Cited by 15 publications

References 4 publications

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

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

Influence of Phenylbutazone on the Phagocytic and Bactericidal Activities of Neutrophil Granulocytes

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

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