Abstract
In vaccine development, especially against pathogenic encapsulated extracellular bacteria, functional assays such as the opsonophagocytosis assay (OPA) are preferred to ELISA titers for evaluating protection against infection. Such assays are normally performed using phagocytic cell lines or purified cell types, which underestimate the complexity of blood bactericidal activity. Here, we describe an OPA using murine whole-blood as effector cells, in a small format (0.2 ml), which requires small quantities of sera (80 μl or less) from immunized individuals. Easy to develop and perform, this OPA can be readily adapted to various pathogens and could be used to evaluate sera from human or animal clinical trials of carbohydrate-based vaccines.
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References
Plotkin SA (2001) Immunologic correlates of protection induced by vaccination. Pediatr Infect Dis J 20:63–75
Plotkin SA (2008) Vaccines: correlates of vaccine-induced immunity. Clin Infect Dis 47:401–409
Plotkin SA (2010) Correlates of protection induced by vaccination. Clin Vaccine Immunol 17:1055–1065
Song JY, Moseley MA, Burton RL et al (2013) Pneumococcal vaccine and opsonic pneumococcal antibody. J Infect Chemother 19:412–425
Abbas AK, Lichtman AH, Pillai S (2007) Effector mechanisms of humoral immunity. In: Schmidtt W (ed) Cellular and Molecular Immunology, 6th edn. Saunders, Philadelphia, PA, pp 321–348
Underhill DM, Ozinsky A (2002) Phagocytosis of microbes: complexity in action. Annu Rev Immunol 20:825–852
Ricklin D, Hajishengallis G, Yang K et al (2010) Complement: a key system for immune surveillance and homeostasis. Nat Immunol 11:785–797
Guilliams M, Bruhns P, Saeys Y et al (2014) The function of Fcg receptors in dendritic cells and macrophages. Nat Rev Immunol 14:94–108
Borrow R, Balmer P, Miller E (2005) Meningococcal surrogates of protection–serum bactericidal antibody activity. Vaccine 23:2222–2227
Martin D, McCallum L, Glennie A et al (2005) Validation of the serum bactericidal assay for measurement of functional antibodies against group B meningococci associated with vaccine trials. Vaccine 23:2218–2221
Balmer P, Borrow R (2004) Serologic correlates of protection for evaluating the response to meningococcal vaccines. Expert Rev Vaccines 3:77–87
Groves E, Dart AE, Covarelli V et al (2008) Molecular mechanisms of phagocytic uptake in mammalian cells. Cell Mol Life Sci 65:1957–1976
Ison CA, Anwar N, Cole MJ et al (1999) Assessment of immune response to meningococcal disease: comparison of a whole-blood assay and the serum bactericidal assay. Microb Pathog 27:207–214
Tan TQ (2012) Pediatric invasive pneumococcal disease in the United States in the era of pneumococcal conjugate vaccines. Clin Microbiol Rev 25:409–419
Goyette-Desjardins G, Auger JP, Xu J et al (2014) Streptococcus suis, an important pig pathogen and emerging zoonotic agent-an update on the worldwide distribution based on serotyping and sequence typing. Emerg Microbes Infect 3:e45
Acknowledgement
This work was mainly supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) through a grant to MS (#342150). Partial contribution was also provided by Canadian Institutes of Health Research grant to MS and RR. The authors wish to thank Dr. Mario Feldman (University of Alberta) and Dr. Marcelo Gottschalk (University of Montreal) for their generous gifts of reference antisera and antibodies against S. pneumoniae and S. suis, respectively.
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Goyette-Desjardins, G., Roy, R., Segura, M. (2015). Murine Whole-Blood Opsonophagocytosis Assay to Evaluate Protection by Antibodies Raised Against Encapsulated Extracellular Bacteria. In: Lepenies, B. (eds) Carbohydrate-Based Vaccines. Methods in Molecular Biology, vol 1331. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2874-3_6
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DOI: https://doi.org/10.1007/978-1-4939-2874-3_6
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2873-6
Online ISBN: 978-1-4939-2874-3
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