Formyl Met-Leu-Phe-Stimulated FPR1 Phosphorylation in Plate-Adherent Human Neutrophils: Enhanced Proteolysis but Lack of Inhibition by Platelet-Activating Factor
Joint Authors
Jesaitis, Algirdas J.
Gripentrog, Jeannie
Voyich, Jovanka M.
Source
Journal of Immunology Research
Issue
Vol. 2018, Issue 2018 (31 Dec. 2018), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2018-01-24
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
N-formyl-Met-Leu-Phe (fMLF) is a model PAMP/DAMP driving human PMN to sites of injury/infection utilizing the GPCR, FPR1.
We examined a microtiter plate format for measurement of FPR1 phosphorylation in adherent PMN at high densities and found that a new phosphosensitive FPR1 fragment, 25K-FPR1, accumulates in SDS-PAGE extracts.
25K-FPR1 is fully inhibited by diisopropylfluorophosphate PMN pretreatment but is not physiologic, as its formation failed to be significantly perturbed by ATP depletion, time and temperature of adherence, or adherence mechanism.
25K-FPR1 was minimized by extracting fMLF-exposed PMN in lithium dodecylsulfate at 4°C prior to reduction/alkylation.
After exposure of adherent PMN to a 5 log range of PAF before or after fMLF, unlike in suspension PMN, no inhibition of fMLF-induced FPR1 phosphorylation was observed.
However, PAF induced the release of 40% of PMN lactate dehydrogenase, implying significant cell lysis.
We infer that PAF-induced inhibition of fMLF-dependent FPR1 phosphorylation observed in suspension PMN does not occur in the unlysed adherent PMN.
We speculate that although the conditions of the assay may induce PAF-stimulated necrosis, the cell densities on the plates may approach levels observed in inflamed tissues and provide for an explanation of PAF’s divergent effects on FPR1 phosphorylation as well as PMN function.
American Psychological Association (APA)
Jesaitis, Algirdas J.& Gripentrog, Jeannie& Voyich, Jovanka M.. 2018. Formyl Met-Leu-Phe-Stimulated FPR1 Phosphorylation in Plate-Adherent Human Neutrophils: Enhanced Proteolysis but Lack of Inhibition by Platelet-Activating Factor. Journal of Immunology Research،Vol. 2018, no. 2018, pp.1-11.
https://search.emarefa.net/detail/BIM-1192056
Modern Language Association (MLA)
Jesaitis, Algirdas J.…[et al.]. Formyl Met-Leu-Phe-Stimulated FPR1 Phosphorylation in Plate-Adherent Human Neutrophils: Enhanced Proteolysis but Lack of Inhibition by Platelet-Activating Factor. Journal of Immunology Research No. 2018 (2018), pp.1-11.
https://search.emarefa.net/detail/BIM-1192056
American Medical Association (AMA)
Jesaitis, Algirdas J.& Gripentrog, Jeannie& Voyich, Jovanka M.. Formyl Met-Leu-Phe-Stimulated FPR1 Phosphorylation in Plate-Adherent Human Neutrophils: Enhanced Proteolysis but Lack of Inhibition by Platelet-Activating Factor. Journal of Immunology Research. 2018. Vol. 2018, no. 2018, pp.1-11.
https://search.emarefa.net/detail/BIM-1192056
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1192056