Chemiluminescence in microamounts of whole blood for investigation of the human phagocyte oxidative metabolism function

Descamps‐Latscha, Béatrice; Nguyen, A.T.; Golub, Robert M.; Feuillet‐Fieux, Marie‐Noëlle

doi:10.1016/0769-2625(82)90047-2

Cited by 33 publications

(11 citation statements)

References 16 publications

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“…Under the conditions of testing and especially at this 10 Ϫ3 blood dilution, both the quenching effect of red blood cells on luminescence photometric measurement and the interference of plasma factors can be ruled out, and luminescence is proportional to the number of phagocytes [30,31]. Luminescence activity was thus normalized with respect to total phagocyte count per microliter (neutrophils, basophils, eosinophils, and monocytes per microliter of blood, with specific activity expressed as counts/20 min/phagocyte) [19].…”

Section: Methodsmentioning

confidence: 99%

Priming of Blood Neutrophils in Children with Cystic Fibrosis: Correlation between Functional and Phenotypic Expression of Opsonin Receptors before and after Platelet‐Activating Factor Priming

et al. 1999

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Blood phagocyte opsonin receptor CR1 (CD35) and CR3 (CD11b) functions were examined in cystic fibrosis (CF) patients with endobronchial Staphylococcus aureus or Pseudomonas aeruginosa chronic infection, CF patients without infection, heterozygous, non-CF patients with chronic pulmonary infection, and healthy controls. Circulating and platelet-activating factor (PAF)-primed phagocyte luminol luminescence responses to complement-opsonized zymosan were increased in both groups of infected CF and non-CF children relative to uninfected CF children and healthy control children and adults. The ratio between circulating and PAF-primed phagocyte responses was significantly elevated in all children with CF, and in these, the ratio could serve as an indicator of response to antibiotic treatment. The ratios of circulating and PAF-primed phenotypic expression for CR1, CR3, and FcgammaRIII (CD16), but not FcgammaRII (CD32), correlated with the functional ratios. Phagocyte opsonin receptor response capacity might be used for evaluation of inflammation and infection in CF patients.

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Section: Methodsmentioning

confidence: 99%

Priming of Blood Neutrophils in Children with Cystic Fibrosis: Correlation between Functional and Phenotypic Expression of Opsonin Receptors before and after Platelet‐Activating Factor Priming

et al. 1999

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“…Zymosan served as a stimulating agent. The functional state of NG was assessed from the chemilumines-0007-4888/94/0007-0777512.50 " @1995 Plenum Publishing Corporation cence index that reflects the reserve capability of the cells [3]. The chemiluminescence index (Ion) was calculated from the following formula:…”

Section: Methodsmentioning

confidence: 99%

Chemiluminescence and morphometric analysis of neutrophil granulocytes from the blood of patients with purulent-septic complications

Medvetskiĭ¹,

Gindich²

1994

Bull Exp Biol Med

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It is demonstrated that the plasma membrane perimeter and the area of neutrophil gr~u-locytes isolated from the blood of patients with a low level of nonspecific protection are signfficantly greater compared with these parameters in patients with a high level of nonspecific protection. These findings can be used to determine the level of nonspecific protection in patients with purulent-septic complications for correction of therapy. Key Words: neutrophil; chemiluminescence; morphometry; purulent-septic complicationsThe need to reduce purulent-septic complications (PSC) using current methods calls for the development of a complex system for predicting postoperative complications; such a system should include tests reflecting the state of nonspecific protection of the organism. Being under the strict control of different systems of the organism and interacting with a number of cells, neutrophils (NG), the main component of nonspecific protection, participate in the maintenance of homeostasis and recognize and eliminate microorganisms and their metabolites [1,7]. The response of NG to bacterial metabolites and cytokines is accompanied by the activation of membrane structures and by the involvement of oxygen-dependent protective mechanisms against infection [2,6]. Such metabolic alterations occur not only in response to contact with foreign agents but also in response to a wide variety of biologically active substances contributing to the maintenance of homeostasis. It is important to find out how the structure of NG changes in relation to their functional activity. Information regarding the relationship between the changes in volume and functional state of neutrophils can be gleaned from the literature. However, it is scarce, controversial, and incomplete and reflects the alterations in cell shape in response to some stimulating (most frequently chemotactic) agent [4,5,9].We studied how the area occupied by an NG and its nucleus, the perimeters of its plasma and nuclear membranes, and the density of nucleus and cytoplasm change depending on the functional state of the cell. The purpose of the study was to establish morphofunctional characteristics for the determination of the level of nonspecific protection. MATERIALS AND METHODSNeutrophils were isolated from the blood of 39 patients with postoperative PSC. The formation of reactive oxygen species (an integral parameter of the functional state of NG) was measured by the method of luminol-dependent chemiluminescence (CL) in a Delta-300 spectrophotometer. The dynamics of spontaneous and stimulated CL was studied. Zymosan served as a stimulating agent. The functional state of NG was assessed from the chemilumines-

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“…The results of these tests were evaluated by recording both absolute changes in spontaneous and stimulated CL and changes in the chemiluminescence index I01, which reflects the reserve capacities of leukocytes [6] and is defined by the formula: 1o1= (CL~-CL)/CL ,, where CL is the maximal spontaneous CL and CL~ is the~maximal stimulated CL, both expressed in counts per minute (cpm).…”

Section: Materialsand Methodsmentioning

confidence: 99%

Functional activity of human leukocytes exposed to a hypotonic medium

Gindich¹

1994

Bull Exp Biol Med

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This study, in which zymosan-stimulated luminol-dependent chemiluminescence of leukocytes was used to examine the osmotic resistance and functional activity of peripheral blood leukocytes from 23 patients with purulent septic lesions and 11 healthy donors before and after exposure of these cells to a hypotonic medium (0.45% NaC1), showed that this medium stimulated their spontaneous chemiluminescence while reducing their reserve capacities. The effects of the hypotonic medium on cells from the patients were more strongly marked. Key Words: leukocytes; chemiluminescence; hypotonic solutionIn the past few years increasing significance has been attached to evaluating the resistance of neutrophilic granulocytes in various disease states [1,2,5,7,8]. The objective of the study reported here was to explore how hypotonic medium as a nonspeciflc factor that alters leukocyte activity might influence one of the earliest and most characteristic metabolic changes in phagocytes, namely the levels of their oxygen metabolism and their cytolysis. MATERIALSAND METHODSPeripheral blood leukocytes from 23 patients with purulent septic lesions and 11 healthy subjects (donors) were examined. As a nonspecific factor eliciting leukolysis and altering the production of intracellular reactive oxygen species, a 0.45% NaC1 solntionwas used [3], in which the percentage of osmotically resistant leukocytes was calculated by the formula:where A is the baseline (pre-exposure) leukocyte count and B is the leukocyte count after a 30-minute exposure to the hypotonic medium. Leukocytes before and after exposure to this medium (at 37~ were counted conductometrically using a blood analyzer.Morphological control of the changes undergone by formed elements under the influence of the hypotonic medium was carried out m whole blood smears and in smears prepared from blood samples incubated in the hypotonic solution.Functional activity of leukocytes was estimated from changes in spontaneous and zymosan-stimulated luminol-dependent chemiluminescence (CL) of intact cells and cells exposed to the hypotonic medium for 30 rain and then transferred to isotonic conditions through the addition of more NaC1 to the medium. The results of these tests were evaluated by recording both absolute changes in spontaneous and stimulated CL and changes in the chemiluminescence index I01, which reflects the reserve capacities of leukocytes [6] and is defined by the formula: 1o1= (CL~-CL)/CL ,, where CL is the maximal spontaneous CL and CL~ is the~maximal stimulated CL, both expressed in counts per minute (cpm).

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Chemiluminescence in microamounts of whole blood for investigation of the human phagocyte oxidative metabolism function

Cited by 33 publications

References 16 publications

Priming of Blood Neutrophils in Children with Cystic Fibrosis: Correlation between Functional and Phenotypic Expression of Opsonin Receptors before and after Platelet‐Activating Factor Priming

Priming of Blood Neutrophils in Children with Cystic Fibrosis: Correlation between Functional and Phenotypic Expression of Opsonin Receptors before and after Platelet‐Activating Factor Priming

Chemiluminescence and morphometric analysis of neutrophil granulocytes from the blood of patients with purulent-septic complications

Functional activity of human leukocytes exposed to a hypotonic medium

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