Development of an opsonophagocytic killing assay for group a streptococcus

Scott Jones, Nicole J. Moreland, Marta Zancolli, Jeremy Raynes, Jacelyn M.S. Loh, Pierre R. Smeesters, Shiranee Sriskandan, Jonathan R. Carapetis, John D. Fraser, David Goldblatt

Research output: Contribution to journalArticle

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Abstract

Group A Streptococcus (GAS) or Streptococcus pyogenes is responsible for an estimated 500,000 deaths worldwide each year. Protection against GAS infection is thought to be mediated by phagocytosis, enhanced by bacteria-specific antibody. There are no licenced GAS vaccines, despite many promising candidates in preclinical and early stage clinical development, the most advanced of which are based on the GAS M-protein. Vaccine progress has been hindered, in part, by the lack of a standardised functional assay suitable for vaccine evaluation. Current assays, developed over 50 years ago, rely on non-immune human whole blood as a source of neutrophils and complement. Variations in complement and neutrophil activity between donors result in variable data that is difficult to interpret. We have developed an opsonophagocytic killing assay (OPKA) for GAS that utilises dimethylformamide (DMF)-differentiated human promyelocytic leukemia cells (HL-60) as a source of neutrophils and baby rabbit complement, thus removing the major sources of variation in current assays. We have standardised the OPKA for several clinically relevant GAS strain types (emm1, emm6 and emm12) and have shown antibody-specific killing for each emm-type using M-protein specific rabbit antisera. Specificity was demonstrated by pre-incubation of the antisera with homologous M-protein antigens that blocked antibody-specific killing. Additional qualifications of the GAS OPKA, including the assessment of the accuracy, precision, linearity and the lower limit of quantification, were also performed. This GAS OPKA assay has the potential to provide a robust and reproducible platform to accelerate GAS vaccine development.

Original languageEnglish
Pages (from-to)3756-3763
Number of pages8
JournalVaccine
Volume36
Issue number26
DOIs
Publication statusPublished - 18 Jun 2018

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Streptococcus
assays
Vaccines
neutrophils
complement
Neutrophils
vaccines
antibodies
antiserum
Antibodies
Immune Sera
rabbits
Rabbits
Streptococcus pyogenes
dimethylformamide
Dimethylformamide
Proteins
proteins
HL-60 Cells
vaccine development

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Jones, S., Moreland, N. J., Zancolli, M., Raynes, J., Loh, J. M. S., Smeesters, P. R., ... Goldblatt, D. (2018). Development of an opsonophagocytic killing assay for group a streptococcus. Vaccine, 36(26), 3756-3763. https://doi.org/10.1016/j.vaccine.2018.05.056
Jones, Scott ; Moreland, Nicole J. ; Zancolli, Marta ; Raynes, Jeremy ; Loh, Jacelyn M.S. ; Smeesters, Pierre R. ; Sriskandan, Shiranee ; Carapetis, Jonathan R. ; Fraser, John D. ; Goldblatt, David. / Development of an opsonophagocytic killing assay for group a streptococcus. In: Vaccine. 2018 ; Vol. 36, No. 26. pp. 3756-3763.
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Jones, S, Moreland, NJ, Zancolli, M, Raynes, J, Loh, JMS, Smeesters, PR, Sriskandan, S, Carapetis, JR, Fraser, JD & Goldblatt, D 2018, 'Development of an opsonophagocytic killing assay for group a streptococcus' Vaccine, vol. 36, no. 26, pp. 3756-3763. https://doi.org/10.1016/j.vaccine.2018.05.056

Development of an opsonophagocytic killing assay for group a streptococcus. / Jones, Scott; Moreland, Nicole J.; Zancolli, Marta; Raynes, Jeremy; Loh, Jacelyn M.S.; Smeesters, Pierre R.; Sriskandan, Shiranee; Carapetis, Jonathan R.; Fraser, John D.; Goldblatt, David.

In: Vaccine, Vol. 36, No. 26, 18.06.2018, p. 3756-3763.

Research output: Contribution to journalArticle

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AU - Jones, Scott

AU - Moreland, Nicole J.

AU - Zancolli, Marta

AU - Raynes, Jeremy

AU - Loh, Jacelyn M.S.

AU - Smeesters, Pierre R.

AU - Sriskandan, Shiranee

AU - Carapetis, Jonathan R.

AU - Fraser, John D.

AU - Goldblatt, David

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Jones S, Moreland NJ, Zancolli M, Raynes J, Loh JMS, Smeesters PR et al. Development of an opsonophagocytic killing assay for group a streptococcus. Vaccine. 2018 Jun 18;36(26):3756-3763. https://doi.org/10.1016/j.vaccine.2018.05.056