A flow cytometry-based assay to determine the ability of anti-Streptococcus pyogenes antibodies to mediate monocytic phagocytosis in human sera.

Autor: Boero E; GSK Vaccines Institute for Global Health (GVGH), Via Fiorentina 1, 53100 Siena, Italy. Electronic address: elena.x.boero@gsk.com., Carducci M; GSK Vaccines Institute for Global Health (GVGH), Via Fiorentina 1, 53100 Siena, Italy., Keeley AJ; Department of Clinical Research, London School of Hygiene and Tropical Medicine, London WC1E 7HT, UK; Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield S10 2TN, UK; Vaccines and Immunity Theme, Medical Research Unit the Gambia at the London School of Hygiene and Tropical Medicine, Atlantic Boulevard, Fajara, P. O. Box 273, the Gambia., Berlanda Scorza F; GSK Vaccines Institute for Global Health (GVGH), Via Fiorentina 1, 53100 Siena, Italy., Iturriza-Gómara M; GSK Vaccines Institute for Global Health (GVGH), Via Fiorentina 1, 53100 Siena, Italy., Moriel DG; GSK Vaccines Institute for Global Health (GVGH), Via Fiorentina 1, 53100 Siena, Italy., Rossi O; GSK Vaccines Institute for Global Health (GVGH), Via Fiorentina 1, 53100 Siena, Italy.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2024 May; Vol. 528, pp. 113652. Date of Electronic Publication: 2024 Mar 06.
DOI: 10.1016/j.jim.2024.113652
Abstrakt: Streptococcus pyogenes, commonly referred to as Group A Streptococcus (Strep A), causes a spectrum of diseases, with the potential to progress into life-threatening illnesses and autoimmune complications. The escalating threat of antimicrobial resistance, stemming from the prevalent reliance on antibiotic therapies to manage Strep A infections, underscores the critical need for the development of disease control strategies centred around vaccination. Phagocytes play a critical role in controlling Strep A infections, and phagocytosis-replicating assays are essential for vaccine development. Traditionally, such assays have employed whole-blood killing or opsonophagocytic methods using HL-60 cells as neutrophil surrogates. However, assays mimicking Fcγ receptors- phagocytosis in clinical contexts are lacking. Therefore, here we introduce a flow cytometry-based method employing undifferentiated THP-1 cells as monocytic/macrophage model to swiftly evaluate the ability of human sera to induce phagocytosis of Strep A. We extensively characterize the assay's precision, linearity, and quantification limit, ensuring robustness. By testing human pooled serum, the assay proved to be suitable for the comparison of human sera's phagocytic capability against Strep A. This method offers a valuable complementary assay for clinical studies, addressing the gap in assessing FcγR-mediated phagocytosis. By facilitating efficient evaluation of Strep A -phagocyte interactions, it may contribute to elucidating the mechanisms required for the prevention of infections and inform the development of future vaccines and therapeutic advancements against Strep A infections.
Competing Interests: Declaration of competing interest This work was undertaken at the request of and sponsored and funded by GlaxoSmithKline Biologicals SA. GSK Vaccines Institute for Global Health Srl is an affiliate of GlaxoSmithKline Biologicals. EB, MC, MI, FBS, MI, DGM, and OR are employees of the GSK group of companies. FBS, DGM, MC, and OR report ownership of GSK shares/share options. AJK is funded by a Wellcome Trust PhD Training Fellowship for Clinicians (225,467/Z/22/Z).
(Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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