Development of a visual Adhesion/Invasion Inhibition Assay to assess the functionality of Shigella -specific antibodies.

Autor:	Batani G; Monoclonal Antibody Discovery Laboratory, Fondazione Toscana Life Sciences, Siena, Italy., Vezzani G; GlaxoSmithKline (GSK) Vaccines Institute for Global Health (GVGH), Siena, Italy., Lashchuk S; Monoclonal Antibody Discovery Laboratory, Fondazione Toscana Life Sciences, Siena, Italy., Allaoui A; The Microbiology Laboratory, University Mohammed VI Polytechnic, Ben, Guerir, Morocco., Cardamone D; Data Science for Health Laboratory, Fondazione Toscana Life Sciences, Siena, Italy., Raso MM; GlaxoSmithKline (GSK) Vaccines Institute for Global Health (GVGH), Siena, Italy., Boero E; GlaxoSmithKline (GSK) Vaccines Institute for Global Health (GVGH), Siena, Italy., Roscioli E; Monoclonal Antibody Discovery Laboratory, Fondazione Toscana Life Sciences, Siena, Italy., Ridelfi M; Monoclonal Antibody Discovery Laboratory, Fondazione Toscana Life Sciences, Siena, Italy., Gasperini G; GlaxoSmithKline (GSK) Vaccines Institute for Global Health (GVGH), Siena, Italy., Pizza M; Department of Life Sciences, Imperial College, London, United Kingdom., Rossi O; GlaxoSmithKline (GSK) Vaccines Institute for Global Health (GVGH), Siena, Italy., Berlanda Scorza F; GlaxoSmithKline (GSK) Vaccines Institute for Global Health (GVGH), Siena, Italy., Micoli F; GlaxoSmithKline (GSK) Vaccines Institute for Global Health (GVGH), Siena, Italy., Rappuoli R; Fondazione Biotecnopolo di Siena, Siena, Italy., Sala C; Monoclonal Antibody Discovery Laboratory, Fondazione Toscana Life Sciences, Siena, Italy.
Jazyk:	angličtina
Zdroj:	Frontiers in immunology [Front Immunol] 2024 Apr 12; Vol. 15, pp. 1374293. Date of Electronic Publication: 2024 Apr 12 (Print Publication: 2024).
DOI:	10.3389/fimmu.2024.1374293
Abstrakt:	Introduction: Shigella is the etiologic agent of a bacillary dysentery known as shigellosis, which causes millions of infections and thousands of deaths worldwide each year due to Shigella 's unique lifestyle within intestinal epithelial cells. Cell adhesion/invasion assays have been extensively used not only to identify targets mediating host-pathogen interaction, but also to evaluate the ability of Shigella -specific antibodies to reduce virulence. However, these assays are time-consuming and labor-intensive and fail to assess differences at the single-cell level. Objectives and Methods: Here, we developed a simple, fast and high-content method named visual Adhesion/Invasion Inhibition Assay (vAIA) to measure the ability of anti- Shigella antibodies to inhibit bacterial adhesion to and invasion of epithelial cells by using the confocal microscope Opera Phenix. Results: We showed that vAIA performed well with a pooled human serum from subjects challenged with S. sonnei and that a specific anti-IpaD monoclonal antibody effectively reduced bacterial virulence in a dose-dependent manner. Discussion: vAIA can therefore inform on the functionality of polyclonal and monoclonal responses thereby supporting the discovery of pathogenicity mechanisms and the development of candidate vaccines and immunotherapies. Lastly, this assay is very versatile and may be easily applied to other Shigella species or serotypes and to different pathogens. Competing Interests: GV, MMR, EB, GG, FBS, FM, and OR are employees of the GSK group of companies. GSK Vaccines Institute for Global Health S.r.l. is an affiliate of GlaxoSmithKline Biologicals SA. FM, FB, OR, and MP report ownership of shares/share options. EB and GV are participating in a Postdoctoral fellowship funded by Wellcome Trust. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2024 Batani, Vezzani, Lashchuk, Allaoui, Cardamone, Raso, Boero, Roscioli, Ridelfi, Gasperini, Pizza, Rossi, Berlanda Scorza, Micoli, Rappuoli and Sala.)
Databáze:	MEDLINE
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