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An Opsonophagocytic Killing Assay for the Evaluation of Group A Streptococcus Vaccine Antisera

McGregor, R; Jones, S; Jeremy, RM; Goldblatt, D; Moreland, NJ; (2020) An Opsonophagocytic Killing Assay for the Evaluation of Group A Streptococcus Vaccine Antisera. Methods in Molecular Biology , 2136 pp. 323-335. 10.1007/978-1-0716-0467-0_26. Green open access

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Abstract

Group A Streptococcus (GAS) is a major cause of global mortality, yet there are no licensed GAS vaccines. Vaccine progress has been hampered, in part, by a lack of standardized assays able to quantify antibody function in test antisera. The most widely used assay was developed over 50 years ago by Rebecca Lancefield and relies on human whole blood as a source of complement and neutrophils. Recently, an opsonophagocytic killing (OPK) assay has been developed for GAS by adapting the OPK methods utilized in Streptococcus pneumoniae vaccine testing. This assay uses dimethylformamide (DMF)-differentiated human promyelocytic leukemia cells (HL-60 cells) as a source of neutrophils and baby rabbit complement, thus removing the major sources of variation in the Lancefield assays. This protocol outlines methods for performing a GAS OPK assay including titering test sera to generate an opsonic index. This in vitro assay could aid in selecting vaccine candidates by demonstrating whether candidate-induced antibodies lead to complement deposition and opsonophagocytic killing.

Type: Article
Title: An Opsonophagocytic Killing Assay for the Evaluation of Group A Streptococcus Vaccine Antisera
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1007/978-1-0716-0467-0_26
Publisher version: http://dx.doi.org/10.1007/978-1-0716-0467-0_26
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Antibody, Complement, Group A Streptococcus, HL-60 cells, Phagocytosis, Vaccine
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/10103530
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