Flow cytometry analysis of the expression of neutrophil FMLP receptors

Allen, Catherine A.; Broom, Murray F.; Chadwick, V. S.

doi:10.1016/0022-1759(92)90246-p

Cited by 28 publications

(12 citation statements)

References 9 publications

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“…Analysis of the fMLP receptor expression was performed based on the principles of the assay described by Allen et al [21]. For staining of fMLP receptors, fLPEP (formyl-Nle-Leu-Phe-Nle-Tyr-Lys; Molecular Probes), a fluorescein-labelled, highly specific binding analogue of fMLP was used, which fluoresces at 520 nm when excited at 488 nm.…”

Section: Expression Of Receptors For Fmlpmentioning

confidence: 99%

Differential effects of clindamycin on neutrophils of healthy donors and septic patients

Wittmann

Arlt

Rothe

et al. 2004

International Immunopharmacology

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Section: Expression Of Receptors For Fmlpmentioning

confidence: 99%

Differential effects of clindamycin on neutrophils of healthy donors and septic patients

Wittmann

Arlt

Rothe

et al. 2004

International Immunopharmacology

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“…For example, priming with LPS can enhance fMLP-induced production of ROS (3)(4)(5). The mechanisms of priming are still poorly understood, but may include enhancement of intracellular signal transduction mechanisms, upregulation of receptors to activating agents (6,7), and/or preliminary assembly of membrane-bound and cytosolic components of NADPH oxidase (which produces ROS).…”

mentioning

confidence: 99%

Mycobacterium tuberculosis19-kDa Lipoprotein Promotes Neutrophil Activation

Neufert¹,

Pai²,

Noss

et al. 2001

The Journal of Immunology

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Certain microbial substances, e.g., LPS, can activate neutrophils or prime them to enhance their response to other activating agents, e.g., fMLP. We investigated the role of the Mycobacterium tuberculosis (MTB) 19-kDa lipoprotein in activation of human neutrophils. MTB 19-kDa lipoprotein initiated phenotypic changes characteristic of neutrophil activation, including down-regulation of CD62 ligand (L-selectin) and up-regulation of CD35 (CR1) and CD11b/CD18 (CR3, Mac-1). In addition, exposure of neutrophils to MTB 19-kDa lipoprotein enhanced the subsequent oxidative burst in response to fMLP as assessed by oxidation of dihydrorhodamine 123 (determined by flow cytometry). LPS also produced these effects with similar kinetics, but an oligodeoxynucleotide containing a CpG motif failed to induce any priming or activation response. Although the effects of LPS required the presence of serum, neutrophil activation by MTB 19-kDa lipoprotein occurred independently of serum factors, suggesting the involvement of different receptors and signaling mechanisms for LPS and MTB 19-kDa lipoprotein. Thus, MTB 19-kDa lipoprotein serves as a pathogen-associated molecular pattern that promotes neutrophil priming and activation.

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“…The ferret PMNs expressed a complete lack of binding to formyl peptide at all concentrations tested (Fig. 2), while human PMNs bound fluorescent-labeled formyl peptide at a concentration (10 À10 M) even lower than previously reported (Allen et al, 1992). FPR, one of the chemotaxis receptors that exists on human neutrophils, belongs to the GPCR (G protein-coupled receptor) family.…”

mentioning

confidence: 69%

“…FPR, one of the chemotaxis receptors that exists on human neutrophils, belongs to the GPCR (G protein-coupled receptor) family. Reports of a FPR binding assay using flow cytometry in human and pig neutrophils (Fletcher et al, 1990;Allen et al, 1992;Graves et al, 1992) found that human, but not porcine, neutrophils bind to the formyl peptide. Thus, the binding reaction to the formyl peptide appears to differ between species.…”

mentioning

confidence: 99%

Chemotactic properties and absence of the formyl peptide receptor in ferret (Mustela putorius furo) neutrophils

Nakata

Otsubo

Kikuchi

et al. 2010

Research in Veterinary Science

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This study describes a chemotaxis assay of ferret polymorphonuclear cells (PMNs). The optimal conditions for this chemotaxis assay were investigated for three chemoattractants: zymosan activated serum (ZAS), recombinant human interleukin-8 (rhIL-8) and N-formyl-Met-Leu- Phe (fMLF). In this study, ferret polymorphonuclear cells (PMNs) reacted to ZAS and rhIL-8, but not fMLF. The optimal concentration of ZAS and rhIL-8 were 5% and 100 ng/ml, respectively. The optimal incubation time of each reagent was 60 min. Due to the lack of response shown from fMLF, the existence of formyl peptide receptors (FPR) on ferret PMNs was investigated by evaluating FPR binding using flow cytometry. The receptor was not detected, implying that ferret neutrophils may lack FPR. This study confirms the fundamental experimental conditions for ferret PMNs chemotaxis and elucidates new findings concerning FPR in ferret neutrophils.

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Flow cytometry analysis of the expression of neutrophil FMLP receptors

Cited by 28 publications

References 9 publications

Differential effects of clindamycin on neutrophils of healthy donors and septic patients

Differential effects of clindamycin on neutrophils of healthy donors and septic patients

Mycobacterium tuberculosis19-kDa Lipoprotein Promotes Neutrophil Activation

Chemotactic properties and absence of the formyl peptide receptor in ferret (Mustela putorius furo) neutrophils

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