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The Development of Immunological Assays to Evaluate the Level and Function of Antibodies Induced by Klebsiella pneumoniae O-Antigen Vaccines

Wagstaffe, Helen R; Johnson, Marina; Osman, Guled; Martin, Patricia; Carranza, Paula; Goldblatt, David; (2023) The Development of Immunological Assays to Evaluate the Level and Function of Antibodies Induced by Klebsiella pneumoniae O-Antigen Vaccines. mSphere 10.1128/msphere.00680-22. (In press). Green open access

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Abstract

Klebsiella pneumoniae, a Gram-negative bacterium, has been listed as a critical pathogen for urgent intervention by the World Health Organization. With no licensed vaccine and increasing resistance to antibiotics, Klebsiella pneumoniae causes a high incidence of hospital- and community-acquired infections. Recently, there has been progress in anti-Klebsiella pneumoniae vaccine development, which has highlighted the lack of standardized assays to measure vaccine immunogenicity. We have developed and optimized methods to measure antibody level and function after vaccination with an in-development Klebsiella pneumoniae O-antigen vaccine. We describe the qualification of a Luminex-based multiplex antibody binding assay and both an opsonophagocytic killing assay and serum bactericidal assay to measure antibody function. Serum from immunized animals were immunogenic and capable of binding to and killing specific Klebsiella serotypes. Cross-reactivity was observed but limited among serotypes sharing antigenic epitopes. In summary, these results demonstrate the standardization of assays that can be used to test new anti-Klebsiella pneumoniae vaccine candidates, which is important for moving them into clinical trials. IMPORTANCE There is no licensed vaccine for the prevention of Klebsiella pneumoniae infections, and increasing levels of antibiotic resistance make this pathogen a high priority for vaccine and therapeutic development. Standardized assays for testing vaccine immunogenicity are paramount for the development of vaccines, and so in this study, we optimized and standardized both antibody-level and function assays for evaluating in-development K. pneumoniae bioconjugate vaccine response in rabbits.

Type: Article
Title: The Development of Immunological Assays to Evaluate the Level and Function of Antibodies Induced by Klebsiella pneumoniae O-Antigen Vaccines
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1128/msphere.00680-22
Publisher version: https://doi.org/10.1128/msphere.00680-22
Language: English
Additional information: Copyright © 2023 Wagstaffe et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license (http://creativecommons.org/licenses/by/4.0/).
Keywords: Klebsiella pneumoniae, vaccine, immunogenicity, multiplex assay, opsonophagocytic killing assay, functional assay
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Infection, Immunity and Inflammation Dept
URI: https://discovery.ucl.ac.uk/id/eprint/10167182
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