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

Biology

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