Kinetics of staphylococcal opsonization, attachment, ingestion and killing by human polymorphonuclear leukocytes: A quantitative assay using [3H] thymidine labeled bacteria

Verhoef, J.; Peterson, Phillip K.; Quie, Paul G.

doi:10.1016/0022-1759(77)90141-7

Cited by 226 publications

(103 citation statements)

References 5 publications

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“…Bacteria were grown overnight at 37°C in 5 ml of MuellerHinton broth (Difco Laboratories, Detroit, MI) to which .02 pCi of [3H]thymidine (specific activity 5 Ci/mmol; Amersham, UK.) had been added (21). Bacterial suspensions were prepared as described previously (8) and were opsonized in diluted pooled bovine serum by incubating on a rotator for 30 min at 37°C.…”

Section: Binding and Ingestion Asmymentioning

confidence: 99%

Preinfection Functions of Blood Polymorphonuclear Leukocytes and the Outcome of Experimental Escherichia coli Mastitis in the Cow

Lohuis¹,

Schukken²,

Henricks³

et al. 1990

Journal of Dairy Science

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The relationship between preinfection functions of blood neutrophils and outcome of experimental Escherichia coli mastitis was studied in 11 cows. Random migration, chemotaxis, phagocytosis, and chemiluminescence by neutrophils were determined in white blood cell suspensions, and in purified neutrophil suspensions. The course of E. coli mastitis (104 E. coli 0:157 in rear quarters) was monitored using clinical parameters, counts of E. coli in mastitic secretion, and milk production. Regressions were calculated for areas under curves of these parameters and preinfection activities of neutrophils. Chemiluminescence by nonstimulated neutrophils in white blood cell suspensions was negatively correlated with counts of E. coli in secretion and with losses in milk production. The chemotactic differential in white blood cell suspensions minus the chemotactic differential in purified suspensions of neutrophils referred to as delta varied from -.66 to +.50, indicating, respectively, inhibition and stimulation of chemotactic activity of neutrophils in white blood cell suspensions. Delta correlated negatively with counts of E. coli in mastitis secretion, inhibition of the amplitude of rumen contractions, and losses in milk production. We hypothesize that a factor in white blood cell suspensions may be involved in the down-regulation of the migratory response of neutrophils during E. coli mastitis.

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Section: Binding and Ingestion Asmymentioning

confidence: 99%

Preinfection Functions of Blood Polymorphonuclear Leukocytes and the Outcome of Experimental Escherichia coli Mastitis in the Cow

Lohuis¹,

Schukken²,

Henricks³

et al. 1990

Journal of Dairy Science

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“…Phagocytosis was assayed as previously described (Verhoef et al, 1977). Briefly, 0.3 ml of the suspension of opsonised [3H]-thymidine-labelled bacteria (5 x 1 O8 cfu/ ml) was added to 0.3 ml of PMNL (lo7 cells/ml) in polypropylene vials (Biovials, Beckman Instruments Inc., Fullerton, CA).…”

Section: Bacterial Uptakementioning

confidence: 99%

Bactericidal, Bacteriolytic and Opsonic Activity of Human Serum Against Escherichia Coli

Rozenberg-Arska

Porsius

Jaarsma

et al. 1986

Journal of Medical Microbiology

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Summary. The effect of human serum on Escherichia coli was studied with serumsensitive and serum-resistant strains. The bactericidal effect of human serum on serumsensitive strains of E. coli depended on the activation of the classical complement pathway. The role of activation of the alternative pathway was less important. After incubation in sub-bactericidal concentrations of serum these strains were also easily phagocytosed by polymorphonuclear leukocytes (PMNL). Strains of E. coli of certain 0-types required not only an intact classical pathway but also the presence of specific antibodies for effective killing by serum and effective phagocytosis by PMNL, despite rapid activation of complement and rapid deposition of C3 on the bacterial surface in the absence of antibody. Capsulate strains O l K l and 078K80 resisted the bactericidal effect of serum even in the presence of specific antibodies; phagocytosis by PMNL only occurred after opsonisation with specific antibodies.

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“…Normal phagocytes rapidly engulf bacteria and yeasts after adherence (18,23), an observation that has led to the omission of differentiation between ingested and adherent particles in phagoctosis assays (14). The small differences in the percentage of green-positive cells, before and after quenching in our experiments, support these findings and would allow assessment of phagocytosis as determined with t%ED50 for FITC-labeled bacteria only (i.e., without adding EB).…”

Section: Discussionmentioning

confidence: 99%

Quantification of Phagocytosis in Human Neutrophils by Flow Cytometry

Heinzelmann

Gardner

Mercer-Jones

et al. 1999

Microbiology and Immunology

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Phagocytosis represents a central element of the host response to microbial invasion. We describe a flow cytometric method for measuring the kinetics of phagocytosis of two bacteria by human polymorphonuclear leukocytes (PMNs). Over a 60-min period, isolated human PMNs were exposed to Staphylococcus aureus (rapidly phagocytosed) and Klebsiella pneumoniae (slowly phagocytosed). This method distinguished adherent from ingested bacteria by quenching fluorescein isothiocyanate-labeled extracellular bacteria with ethidium bromide. This further allowed the exclusion of dead, highly permeable, and subsequently bright-red fluorescent PMNs. Our experiments with two different bacteria, various PMN-to-bacteria ratios (1:1, 1:10, 1:100), and different individuals proved that 1) flow cytometric analysis is accurate and useful for characterizing phagocytosis, 2) adherent bacteria can be distinguished from ingested bacteria after quenching with ethidium bromide, and that 3) phagocytosis kinetics of two bacteria with different onsets of phagocytosis can be determined by flow cytometry and the assessment of a score that quantifies phagocytosis.

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Kinetics of staphylococcal opsonization, attachment, ingestion and killing by human polymorphonuclear leukocytes: A quantitative assay using [3H] thymidine labeled bacteria

Cited by 226 publications

References 5 publications

Preinfection Functions of Blood Polymorphonuclear Leukocytes and the Outcome of Experimental Escherichia coli Mastitis in the Cow

Preinfection Functions of Blood Polymorphonuclear Leukocytes and the Outcome of Experimental Escherichia coli Mastitis in the Cow

Bactericidal, Bacteriolytic and Opsonic Activity of Human Serum Against Escherichia Coli

Quantification of Phagocytosis in Human Neutrophils by Flow Cytometry

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