Evaluation of a fluorochrome assay for assessing the bactericidal activity of neutrophils in human phagocyte dysfunctions

Bellinati-Pires, Raquel; Melki, S.E.; Colletto, G M D D; Carneiro-Sampaio, Magda

doi:10.1016/0022-1759(89)90395-5

Cited by 48 publications

(37 citation statements)

References 14 publications

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“…Phagocytosis and bacterial killing by neutrophils and monocytes were assessed by a fluorochrome technique [17,18], using acridine orange staining to determine living (green) and non-living (red/yellow) bacteria. Phagocytosis index (PI), and percent killed bacteria (%KB) were calculated as in Bellinati-Pires et al [18].…”

Section: Phagocytosis and Bacterial Killingmentioning

confidence: 99%

Phagocytosis and killing of epidemic methicillin-resistant Staphylococcus aureus by human neutrophils and monocytes

Salgado¹,

Pignatari²,

Bellinati-Pires³

2004

Braz J Infect Dis

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Staphylococcus aureus is a pathogen that has been associated with nosocomial infections since the preantibiotic era. Since the introduction of antibiotics in medical practice in the 1940's, methicillin-resistant S. aureus (MRSA) strains have been emerging in various parts of the world. In view of the important role of the phagocytic system in the defense against this bacteria, we decided to study phagocytosis by neutrophils and monocytes of an epidemic MRSA strain in São Paulo, Brazil, in comparison with methicillin-sensitive strains. Complement system opsonins are fundamental for efficient ingestion of the resistant and sensitive strains by both types of phagocytes. We found no association of the opsonic requirement of the MRSA strain with the multiresistance phenotype. On the other hand, the MRSA strain was found to be more resistant to the effector mechanisms of neutrophils than both sensitive strains when opsonized with fresh serum, despite the phagocytosis results. This fact suggests that the intracellular killing of S. aureus is an additional parameter of bacterial virulence, but new approaches must be implemented to study the interactions of this MRSA strain with phagocytes in order to investigate the possible factors involved in its behavior in response to neutrophil effector mechanisms.

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Section: Phagocytosis and Bacterial Killingmentioning

confidence: 99%

Phagocytosis and killing of epidemic methicillin-resistant Staphylococcus aureus by human neutrophils and monocytes

Salgado¹,

Pignatari²,

Bellinati-Pires³

2004

Braz J Infect Dis

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“…A modification of the method described by Bellinati-Pires et al 6 was used to examine the influence of propofol on phagocytosis and killing of bacteria by PMNL. After having been cultured on blood agar plates at 37~ for 24 hr, Staphylococcus aureus (serovar K:807) and Escherichia coli (serovar 0:6) were inoculated to Todd Hewitt nutrient broth (Biotest, Dreieich) and incubated at 37~ for 24 hr.…”

Section: Phagocytosis and Bactericidal Activitymentioning

confidence: 99%

Propofol inhibits phagocytosis and killing ofStaphylococcus aureus andEscherichia coli by polymorphonuclear leukocytesin vitro

1994

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Polymorphonuclear leukocytes (PMNL) About a century ago Platania J observed an intensification of bacterial infections in laboratory animals caused by anaesthetic drugs. He regarded a disturbance of the nervous system as the reason for his discovery. This opinion, however, was not shared by all scientists, some of whom had the idea that anaesthetics might inhibit the bactericidal effect of the blood. A few years later Graham 2 discovered a suppression of phagocytosis of bacteria by polymorphonuclear leukocytes (PMNL) caused by ether. Since then, several studies have been conducted concerning the influence of anaesthetic agents on PMNL function. A current survey was published by Kress and Eberlein. 3 However, there are few findings regarding the effect of propofol. In particular, its influence on the confrontation between PMNL and human pathogenic bacteria has not been examined up to now. This induced us to investigate the effects of propofol and its solvent intralipid on phagocytosis and killing of Staphylococcus aureus and Escherichia coli, in vitro. MethodsPermission to conduct the study was obtained from the Ethics Committee of the Medical Faculty of JustusLiebig-University Giessen. Blood samplesVenous blood samples (1.5 mg ethylenediaminetetraaceticdipotassium ml-I; Sigma, Deisenhofen) were obtained from ten healthy male volunteers (age: 29 +4 yr; body weight: 75 4-8 kg; height: 1.80 + 0.05 m (mean and SD). Isolation of PMNLThe PMNL were isolated using a method described by CAN J ANAESTH 1994 / 41:5 / pp 446--9

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“…A volume of 5 ml of the cell suspension was pipetted onto microscope slides and examined at Â1000 final magnification with a Zeiss ultraviolet confocal laser-scanning microscope (Zeiss, model LSM 510; Jena, Thuringia, Germany). Bacteria were scored on the basis of green (live bacteria) or red/yellow (dead bacteria) fluorescence [10].…”

Section: Methodsmentioning

confidence: 99%

Human IgG but not IgM Antibodies can Protect Mice from the Challenge with Live O6 Escherichia coli

et al. 2005

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We evaluated the ability of human anti-lipopolysaccharide (LPS) O6 immunoglobulin G (IgG) and IgM antibodies to protect mice challenged with Escherichia coli serotype O6:K2ac. Purified whole IgG, commercial gammaglobulin, whole IgM-effluent, pool of normal human serum (NHS), agammaglobulinaemic serum (test groups) or phosphate-buffered saline (control group) was injected into adult male 18 h before a challenge with viable O6 E. coli. The mortality rate was assessed over a period of 72 h. To determine the opsonic and phagocytic activity of the antibody isotypes, we incubated peritoneal macrophages from the control and test groups collected at different times after challenge with the live bacteria with acridine orange for fluorescent analysis.

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Evaluation of a fluorochrome assay for assessing the bactericidal activity of neutrophils in human phagocyte dysfunctions

Cited by 48 publications

References 14 publications

Phagocytosis and killing of epidemic methicillin-resistant Staphylococcus aureus by human neutrophils and monocytes

Phagocytosis and killing of epidemic methicillin-resistant Staphylococcus aureus by human neutrophils and monocytes

Propofol inhibits phagocytosis and killing ofStaphylococcus aureus andEscherichia coli by polymorphonuclear leukocytesin vitro

Human IgG but not IgM Antibodies can Protect Mice from the Challenge with Live O6 Escherichia coli

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