Development of a serological assay to predict antibody bactericidal activity against non-typeable Haemophilus influenzae

BMC Microbiology

Silvia Rossi-Paccani¹  Marco Soriani¹  Mariagrazia Pizza¹  Beatrice Aricò¹  Roberto Petracca¹  Sara Marchi¹  Greco Alessandra¹  Buket Baddal¹  Giuseppe Ercoli² 

[1] Novartis Vaccines & Diagnostics s.r.l. (a GSK company), Via Fiorentina 1, Siena, 53100, Italy;University of Leicester, Department of Genetics, Adrian building, Leicester, UK

关键词: Vaccine;    Complement;    Antibody;    Serum bactericidal assay;    Non-typeable Haemophilus influenzae;   

Others  :  1212065
DOI  :  10.1186/s12866-015-0420-x

 received in 2015-01-08, accepted in 2015-04-01,  发布年份 2015

【 摘 要 】

Background

Non-typeable Haemophilus influenzae (NTHi) is a Gram negative microorganism residing in the human nasopharyngeal mucosa and occasionally causing infections of both middle ear and lower respiratory airways. A broadly protective vaccine against NTHi has been a long-unmet medical need, as the high genetic variability of this bacterium has posed great challenges.

Results

In this study, we developed a robust serum bactericidal assay (SBA) to optimize the selection of protective antigens against NTHi. SBA takes advantage of the complement-mediated lysis of bacterial cells and is a key in vitro method for measuring the functional activity of antibodies. As a proof of concept, we assessed the bactericidal activity of antibodies directed against antigens known to elicit a protective response, including protein D used as carrier protein in the Synflorix pneumococcal polysaccharide conjugate vaccine. Prior to SBA screening, the accessibility of antigens to antibodies and the capacity of the latter to induce C3 complement deposition was verified by flow cytometry. Using baby rabbit serum as a source of complement, the proposed assay not only confirmed the bactericidal activity of the antibodies against the selected vaccine candidates, but also showed a significant reproducibility.

Conclusions

Considering the rapidity and cost-effectiveness of this novel SBA protocol, we conclude that it is likely to become an important tool to prove the capability of antibodies directed against recombinant antigens to induce NTHi in vitro killing and to both select new protective vaccine candidates, and predict vaccine efficacy.

【 授权许可】

   
2015 Ercoli et al.; licensee BioMed Central.

【 图 表 】

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