Measurement of phagocytosis and oxidative burst of canine neutrophils: high variation in healthy dogs

Eickhoff, Sandra; Mironowa, Lilia; Carlson, Regina; Leibold, W.; Tipold, Andrea

doi:10.1016/j.vetimm.2004.04.011

Cited by 18 publications

(4 citation statements)

References 23 publications

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“…Surprisingly, oxidative burst and phagocytic activities of the phagocytes were not different between dogs with CPE and the control dogs, likely for multifactorial reasons. First, the oxidative burst and phagocytic activities of neutrophils in healthy dogs varies widely between individuals, 14 which was also evident in our study in both health and disease. Second, in humans, systemic inflammation induces functional heterogeneity among neutrophil phenotypes such that cells with differing functional capacities are simultaneously present in circulation.…”

Section: Discussionsupporting

confidence: 70%

“…To reduce the analytical variation of the assay, only the principal investigator gated the phagocyte populations, as recommended. 14 The results were reported as the percentage of fluorescence-positive cells, which equates to the proportion of phagocytes containing rhodamine 123 or FITC-labeled E. coli as a measure of oxidative burst or phagocytic activity, respectively. The mean fluorescence intensity (MFI) was also reported and equates to the average magnitude of ROS production per cell when stimulated by E. coli or PMA, or the average number of E. coli phagocytized per cell.…”

Section: Methodsmentioning

confidence: 99%

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Oxidative burst and phagocytic activity of phagocytes in canine parvoviral enteritis

et al. 2021

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Canine parvoviral enteritis (CPE) is a severe disease characterized by systemic inflammation and immunosuppression. The function of circulating phagocytes (neutrophils and monocytes) in affected dogs has not been fully investigated. We characterized the functional capacity of canine phagocytes in CPE by determining their oxidative burst and phagocytic activities using flow cytometry. Blood was collected from 28 dogs with CPE and 11 healthy, age-matched, control dogs. Oxidative burst activity was assessed by stimulating phagocytes with opsonized Escherichia coli or phorbol 12-myristate 13-acetate (PMA) and measuring the percentage of phagocytes producing reactive oxygen species and the magnitude of this production. Phagocytosis was measured by incubating phagocytes with opsonized E. coli and measuring the percentage of phagocytes containing E. coli and the number of bacteria per cell. Complete blood counts and serum C-reactive protein (CRP) concentrations were also determined. Serum CRP concentration was negatively and positively correlated with segmented and band neutrophil concentrations, respectively. Overall, no differences in phagocyte function were found between dogs with CPE and healthy control dogs. However, infected dogs with neutropenia or circulating band neutrophils had decreased PMA-stimulated oxidative burst activity compared to healthy controls. Additionally, CPE dogs with neutropenia or circulating band neutrophils had decreased PMA- and E. coli–stimulated oxidative burst activity and decreased phagocytosis of E. coli compared to CPE dogs without neutropenia or band neutrophils. We conclude that phagocytes have decreased oxidative burst and phagocytic activity in neutropenic CPE dogs and in CPE dogs with circulating band neutrophils.

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Section: Discussionsupporting

confidence: 70%

Section: Methodsmentioning

confidence: 99%

Oxidative burst and phagocytic activity of phagocytes in canine parvoviral enteritis

et al. 2021

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“…The cell suspension was then immediately analyzed with flow cytometry. 22,23 Monochlorobimane Assay of GSH Monochlorobimane (mBCl) e is a nonfluorescent molecule that easily crosses cell membranes and forms a conjugation product with intracellular GSH. This conjugation product (mBCl-GSH) is fluorescent; therefore, the subsequent increase in fluorescence detected by flow cytometry represents the concentration of intracellular GSH.…”

Section: Neutrophil Phagocytosis Assaymentioning

confidence: 99%

Neutrophil Function in Septic Dogs

Webb¹,

McCord²,

Dow³

2007

Veterinary Internal Medicne

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Background: Leukocytes appear to enter a hypo-inflammatory state in human patients with a severe bacterial infection, whereas, in swine, intra-abdominal sepsis produces an initial increase and subsequent decrease in neutrophil Fc receptormediated phagocytosis.Hypothesis: Impaired neutrophil function (hypo-inflammatory state) develops in dogs with sepsis. Animals: Thirteen adult client-owned dogs that developed clinical signs consistent with sepsis were assessed for evidence of neutrophil dysfunction. These results were compared with those of 12 healthy dogs.Methods: Flow cytometry combined with the appropriate fluorescent markers allowed for quantification of the phagolysosomal oxidative burst after Fc receptor-mediated phagocytosis of immune complexes, neutrophil phagocytosis of opsonized Escherichia coli, and the intracellular concentration of reduced glutathione as a measure of oxidative stress.Results: The phagolysosomal oxidative burst after Fc receptor-mediated phagocytosis was significantly lower in neutrophils from septic dogs (mean fluorescence intensity 6 standard deviation; 118 6 13 and 165 6 27 for septic and control dogs, respectively, p 5 0.001), although the phagocytosis of opsonized E coli was significantly increased (155 6 74 and 77 6 44 for septic and control dogs, respectively, p 5 0.004). Intracellular reduced glutathione was not significantly different in neutrophils from septic and healthy control dogs.Conclusions: An important component of neutrophil function is decreased in septic dogs. The diminished oxidative burst after Fc receptor-mediated phagocytosis in neutrophils from septic dogs could hinder the ability of the innate immune system to clear bacterial infections or it might help protect these patients from the systemic consequences of infection.

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“…Neutrophils and monocytes are the first line of defense against bacterial infections (Underhill and Ozinsky 2002) and act to defend the host by chemotaxis, phagocytosis, releasing lysosomal enzymes and generating reactive oxygen metabolites (Eickhoff et al 2004;Underhill and Ozinsky 2002). The first step in this process is the recognition of and attachment to the target.…”

Section: Introductionmentioning

confidence: 99%

Zinc increases the phagocytic capacity of canine peripheral blood phagocytes in vitro

2008

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Zinc is a trace element that plays a central role in the immune system. In the present study, the effect of zinc on the phagocytic capacity of canine peripheral blood phagocytes was examined in vitro by flow cytometry. Zinc was used at a concentration of 100 microM, which preserved cell viability. Treatment with zinc did not directly affect the phagocytic capacity of peripheral blood polymorphonuclear neutrophils (PMN) and mononuclear cells (PBMC). However, it did directly enhance the phagocytic capacity of peripheral blood monocyte-rich cells. Moreover, the phagocytic capacity of PMN and monocyte-rich cells but not PBMC was remarkably enhanced by culture supernatants from PBMC but not PMN treated with zinc. Anti-recombinant canine (rc) tumor necrosis factor-alpha (TNF-alpha) polyclonal antibody (pAb) neutralized the enhancing effect of the culture supernatant from zinc-treated PBMC and this supernatant had higher TNF-alpha levels than the culture supernatant of untreated PBMC. Thus, zinc may stimulate canine PBMC to produce TNF-alpha, which enhances the phagocytic capacity of canine peripheral blood phagocytes.

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Measurement of phagocytosis and oxidative burst of canine neutrophils: high variation in healthy dogs

Cited by 18 publications

References 23 publications

Oxidative burst and phagocytic activity of phagocytes in canine parvoviral enteritis

Oxidative burst and phagocytic activity of phagocytes in canine parvoviral enteritis

Neutrophil Function in Septic Dogs

Zinc increases the phagocytic capacity of canine peripheral blood phagocytes in vitro

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