Chemiluminescence neutrophil

Lucas, M., and Solano, F. (1992). Coelenterazine is a superoxide anion-sensitive chemiluminescent probe its usefulness in the assay of respiratory burst in neutrophils. Anal. Biochem. 206 273-277. 

This is a chronic inflammatory disease, which can affect the gut as well as other organs. There is relatively little information on the role of free radicals in this condition. Neutrophil chemiluminescence was increased in patients with intestinal Behcet s compared to normal controls (Suematsu et al., 1987a) and there is some evidence for endothelial injury by neutrophil-derived oxidants (Niwa et al., 1982). Preliminary studies with lip)osomal-encapsulated SOD demonstrated marked improvement in 12 out of 16 pjatients wdth active Behcet s disease (Niwa etal., 1985). 

Faden, H. and Rossi, T.M. (1985). Chemiluminescent response of neutrophils fixjm patients with inflammatory bowel disease. Dig. Dis. Sci. 30, 139-142. 

Stewart, W. W., Kerr, M. A. (1990). The specificity of the human neutrophil IgA receptor (Fc aR) determined by measurement of chemiluminescence induced by serum or secretory IgAl or IgA2. Immunol. 71, 328-34. 

When neutrophils are stimulated with fMet-Leu-Phe, they generate a bi-phasic luminol chemiluminescence response. The first phase of chemilumi-... 

The molecular species responsible for lucigenin chemiluminescence are not fully defined. Because this probe does not permeate the cell, only secreted oxidants are detected. Several reports have suggested that this probe measures O2 secretion or else H2O2 secretion on the other hand, some evidence indicates that this cannot be so. Certainly, the oxidant(s) detected by lucigenin is dependent on O2 formation, but the kinetics of activated 02" and H2O2 formation by neutrophils do not parallel those of lucigenin chemiluminescence. Thus, this probe is at least an indirect measure of O2 secretion. 

In summary, chemiluminescence is a sensitive, non-invasive technique that can measure reactive oxidant production by small numbers of neutrophils indeed, neutrophil-derived chemiluminescence can be detected in as little as 5 fA of unfractionated human blood. The assay is suitable for automation using either multichannel luminometers or luminescence microtitre plate readers. Many researchers, however, have questioned the usefulness of this technique because of the uncertainty of the nature of the oxidant(s) that are detected. Nevertheless, in view of the recent developments made towards the identification of the oxidants measured and the assay s ability to detect intracellular oxidant production, it is has an important place in the phagocyte research laboratory. 

Dahlgren, C., Aniansson, H., Magnusson, K.-E. (1985). Pattern of formylmethionyl-leucyl-phenylalanine-induced luminol- and lucigenin-dependent chemiluminescence in human neutrophils. Infect. Immun. 47, 326-8. 

Edwards, S. W. (1987). Luminol- and lucigenin-dependent chemiluminescence of neutrophils Role of degranulation. J. Clin. Lab. Immunol. 22, 35-9. 

These approaches have been used to show conclusively that the initial, low formation of DAG that occurs during activation with soluble agonists comes from PLC activity, and that the later, more sustained generation of DAG comes from PLD activity. Such experiments have also shown that primary alcohols can inhibit the activity of the NADPH oxidase under some conditions. When neutrophils are pretreated with cytochalasin B, primary alcohols are potent inhibitors of 02" secretion, and the kinetics of phosphatidic acid formation are rapid, peaking within about 20 s and coinciding with oxidase activation. However, in the absence of cytochalasin B, primary alcohols have little effect on the initiation of O2" secretion, but decrease the duration of oxidase activity they also inhibit the later phase of luminol chemiluminescence, which is largely intracellular, and the kinetics of phosphatidic acid formation closely parallel the kinetics of this intracellular oxidase activity (Fig. 6.20). Thus, in cytochalasin-treated cells, PLD is activated rapidly, and this activation is required for 02" secretion in the absence of cytochalasin, PLD is activated more slowly and its function is not for the activation of the oxidase, but rather for sustained (and intracellular) activity. 

Figure 6.20. Role of phospholipase D in NADPH oxidase activation. In (a) neimophils were preincubated with [3H]-alkyl-lyso-PAF (5 /iCi/ml) for 60 nun at 37 C. The cells were then washed twice with RPMI 1640 medium and finally resuspended at 2 x 10 cells/ ml in the presence ( ) and absence ( ) of 100 mM ethanol. The cells were then stimulated with 1 pM fMet-Leu-Phe and, at time intervals,aliquots were removed for analysis ofphos-phatidic acid ( ) and phosphatidylethanol ( ) by thin layer chromatography (TLC). In (b), neutrophils were incubated in the presence and absence of 10 mM butanol, and luminol chemiluminescence (10 jUM, final concentration of luminol) was measured after stimulation by 1 jUM fMet-Leu-Phe. Source Experiment of Gordon Lowe and Fiona Watson.

Figure 7.6. Activation of neutrophils in unfractionated blood. Unfractionated, whole blood (10 p ) was diluted to 1 ml in RPMI 1640 medium and incubated in the absence (un-primed) or presence (primed) of 50 U/ml GM-CSF for 60 min at 37 °C. After this incubation, luminol was added (to 100 pM, final concentration) and chemiluminescence measured after the addition of 1 pM fMet-Leu-Phe.

Figure 7.12. Role of protein biosynthesis in the ability of neutrophils to generate reactive oxidants. Neutrophils were incubated in the presence (O) and absence ( ) of 30 pg/ml cycloheximide at 37 °C. At time intervals, portions were removed and luminol chemiluminescence measured after stimulation with 1 pM fMet-Leu-Phe. Values presented are a percentage of the control value measured at time zero.

Hyvonen PM, Kowolik MJ. Human neutrophil priming chemiluminescence modified by hydroxyapatite and three bisphosphonates in vitro. J Clin Lab Immunol 1993 40 69-76. 

Babior, B. M. Recent studies on oxygen metabolism in human neutrophils Superoxide and chemiluminescence. In Superoxide and Superoxide Dismutases (Michelson, A. M., McCord, J. M., Fridovich, I., eds.), London-New York-San Francisco, Academic Press, 1977, pp. 271-281 McCord, J. M., Wong, K. Phagocyte-produced free radicals Roles in cytotoxicity and inflammation. In Oxygen Free Radicals and Tissue Damage, Ciba Foundation Symposium 65, Amsterdam-Oxford-New York, Excerpta Medica, 1979, pp. 343-351... 

K. Cantell. 1995. Inhaled recombinant interferon gamma in patients with lung cancer pharmacokinetics and effects on chemiluminescence responses of alveolar macrophages and peripheral blood neutrophils and monocytes. Int. 

The flow of leukocytes was studied in square capillaries fabricated on a Si chip, and sealed with a PDMS or Pyrex cover plate. This capillary size (cross section of 4 pm2) is similar to the diameter of a human blood capillary, but is less than both the average diameter of a leukocyte cell (10 pm) and its nucleus (6 pm). Figure 8.32 shows the difference in the flow behavior of two leukocytes (possibly neutrophils) [1175]. Deformation-induced release of ATP from erythrocytes in PDMS channels was studied. The released ATP was detected by chemiluminescence using the luciferin/luciferase system [169]. 

Weiss M, Buhl R, Birkhahn A, Mirow N, Schneider M, Wemet P (1994a) Do barbiturates and their solutions suppress FMLP-induced neutrophil chemiluminescence Eur J Anaesthesiol 11 371-379. 

Propylene glycol in a concentration of 0.5-1.0% has been shown to inhibit natural cytotoxicity and neutrophil chemiluminescence in human cells in vitro in one study. 

Kinetic analysis of luminol-dependent chemiluminescence demonstrated that a solution of berbamine (20 pM) was found to inhibit the generation of various types of reactive oxygens by guinea-pig neutrophils. The results of these and other experiments suggest that berbamine inhibits the active oxygen generation via the stabilization of plasma membrane and the inhibition of phospholipid-dependent protein kinase (PKC) and NADPH oxidase activation [178]. 

Thompson, P.J., Misso, N.L., Passarelli, M., and Phillips, M.J. 1991. The effect of eicosapentaenoic acid consumption on human neutrophil chemiluminescence. Lipids 26, 1223-1226. 

Belotskii SM, Filiudova OB, Pashutin SB, et al Chemiluminescence of Human Neutrophils as Affected by Opportunistic Microbes. Zh. Mikrobiol. Epidemiol. Immunobiol. 1986 Mar (3) 89-92. 

Gorski P, Tarkowski M, Krakowiak A, et al. 1992. Neutrophil chemiluminescence following exposure to formaldehyde in healthv subjects and in patients with contact dermatitis. Allergol Immunopathol (Madr) 20 20-23. 

Ml. McPhail, L. C., DeChatelet, L. R, and Johnston, R. B., Jr., Generation of chemiluminescence by a particulate fraction isolated from human neutrophils. Analysis of molecular events. J. Clin. Invest. 63, 648-655 (1979). 

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