Evaluation of inactivated Bordetella pertussis as a delivery system for the immunization of mice with Pneumococcal Surface Antigen A

Autoři: Julia T. Castro aff001;  Giuliana S. Oliveira aff001;  Melissa A. Nishigasako aff001;  Anne-Sophie Debrie aff002;  Eliane N. Miyaji aff001;  Alessandra Soares-Schanoski aff001;  Milena A. Akamatsu aff003;  Camille Locht aff002;  Paulo L. Ho aff003;  Nathalie Mielcarek aff002;  Maria Leonor S. Oliveira aff001
Působiště autorů: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil aff001;  Univ. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019 –UMR 8204 –CIIL—Center for Infection and Immunity of Lille, Lille, France aff002;  Seção de Vacinas Aeróbicas, Divisão Bioindustrial, Instituto Butantan, São Paulo, SP, Brazil aff003
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0228055


Pneumococcal Surface Protein A (PspA) has been successfully tested as vaccine candidate against Streptococcus pneumoniae infections. Vaccines able to induce PspA-specific antibodies and Th1 cytokines usually provide protection in mice. We have shown that the whole cell pertussis vaccine (wP) or components from acellular pertussis vaccines, such as Pertussis Toxin or Filamentous Hemagglutinin (FHA), are good adjuvants to PspA, suggesting that combined pertussis-PspA vaccines would be interesting strategies against the two infections. Here, we evaluated the potential of wP as a delivery vector to PspA. Bordetella pertussis strains producing a PspA from clade 4 (PspA4Pro) fused to the N-terminal region of FHA (Fha44) were constructed and inactivated with formaldehyde for the production of wPPspA4Pro. Subcutaneous immunization of mice with wPPspA4Pro induced low levels of anti-PspA4 IgG, even after 3 doses, and did not protect against a lethal pneumococcal challenge. Prime-boost strategies using wPPspA4Pro and PspA4Pro showed that there was no advantage in using the wPPspA4Pro vaccine. Immunization of mice with purified PspA4Pro induced higher levels of antibodies and protection against pneumococcal infection than the prime-boost strategies. Finally, purified Fha44:PspA4Pro induced high levels of anti-PspA4Pro IgG, but no protection, suggesting that the antibodies induced by the fusion protein were not directed to protective epitopes.

Klíčová slova:

Antibodies – Bordetella pertussis – Cloning – Enzyme-linked immunoassays – Pneumococcus – Recombinant proteins – Recombinant vaccines – Vaccines


1. GBD (2018) Estimates of the global, regional, and national morbidity, mortality, and aetiologies of lower respiratory infections in 195 countries, 1990–2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet Infect Dis 18: 1191–1210. doi: 10.1016/S1473-3099(18)30310-4 30243584

2. Yildirim I, Shea KM, Pelton SI (2015) Pneumococcal Disease in the Era of Pneumococcal Conjugate Vaccine. Infect Dis Clin North Am 29: 679–697. doi: 10.1016/j.idc.2015.07.009 26610421

3. Davis SM, Deloria-Knoll M, Kassa HT, O'Brien KL (2013) Impact of pneumococcal conjugate vaccines on nasopharyngeal carriage and invasive disease among unvaccinated people: review of evidence on indirect effects. Vaccine 32: 133–145. doi: 10.1016/j.vaccine.2013.05.005 23684824

4. Weinberger DM, Malley R, Lipsitch M (2011) Serotype replacement in disease after pneumococcal vaccination. Lancet 378: 1962–1973. doi: 10.1016/S0140-6736(10)62225-8 21492929

5. Miyaji EN, Oliveira ML, Carvalho E, Ho PL (2013) Serotype-independent pneumococcal vaccines. Cell Mol Life Sci 70: 3303–3326. doi: 10.1007/s00018-012-1234-8 23269437

6. Ren B, Li J, Genschmer K, Hollingshead SK, Briles DE (2012) The absence of PspA or presence of antibody to PspA facilitates the complement-dependent phagocytosis of pneumococci in vitro. Clin Vaccine Immunol 19: 1574–1582. doi: 10.1128/CVI.00393-12 22855389

7. Shaper M, Hollingshead SK, Benjamin WH Jr., Briles DE (2004) PspA protects Streptococcus pneumoniae from killing by apolactoferrin, and antibody to PspA enhances killing of pneumococci by apolactoferrin [corrected]. Infect Immun 72: 5031–5040. doi: 10.1128/IAI.72.9.5031-5040.2004 15321996

8. Oliveira ML, Miyaji EN, Ferreira DM, Moreno AT, Ferreira PC, Lima FA et al. (2010) Combination of pneumococcal surface protein A (PspA) with whole cell pertussis vaccine increases protection against pneumococcal challenge in mice. PLoS One 5: e10863. doi: 10.1371/journal.pone.0010863 20523738

9. Tostes RO, Rodrigues TC, da Silva JB, Schanoski AS, Oliveira ML, Miyaji EN. (2017) Protection Elicited by Nasal Immunization with Recombinant Pneumococcal Surface Protein A (rPspA) Adjuvanted with Whole-Cell Pertussis Vaccine (wP) against Co-Colonization of Mice with Streptococcus pneumoniae. PLoS One 12: e0170157. doi: 10.1371/journal.pone.0170157 28103277

10. Salcedo-Rivillas C, Debrie AS, Miyaji EN, Ferreira JM Jr., Raw I, Locht C, et al. (2014) Pertussis toxin improves immune responses to a combined pneumococcal antigen and leads to enhanced protection against Streptococcus pneumoniae. Clin Vaccine Immunol 21: 972–981. doi: 10.1128/CVI.00134-14 24807055

11. Akamatsu MA, Nishiyama MY Jr., Morone M, Oliveira UC, Bezerra MF, Sakauchi MA, et al. (2015) Whole-Genome Sequence of a Bordetella pertussis Brazilian Vaccine Strain. Genome Announc 3.

12. Kammoun H, Roux X, Raze D, Debrie AS, De Filette M, Ysenbaert T, et al. (2013) Immunogenicity of live attenuated B. pertussis BPZE1 producing the universal influenza vaccine candidate M2e. PLoS One 8: e59198. doi: 10.1371/journal.pone.0059198 23555631

13. Moreno AT, Oliveira ML, Ferreira DM, Ho PL, Darrieux M, Leite LC, et al. (2010) Immunization of mice with single PspA fragments induces antibodies capable of mediating complement deposition on different pneumococcal strains and cross-protection. Clin Vaccine Immunol 17: 439–446. doi: 10.1128/CVI.00430-09 20089795

14. Ramos CR, Abreu PA, Nascimento AL, Ho PL (2004) A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide. Braz J Med Biol Res 37: 1103–1109. doi: 10.1590/s0100-879x2004000800001 15273812

15. Figueiredo DB, Carvalho E, Santos MP, Kraschowetz S, Zanardo RT, Campani G Jr et al. (2017) Production and purification of an untagged recombinant pneumococcal surface protein A (PspA4Pro) with high-purity and low endotoxin content. Appl Microbiol Biotechnol 101: 2305–2317. doi: 10.1007/s00253-016-7983-9 27889801

16. Briles DE, Hollingshead SK, King J, Swift A, Braun PA, Park, MK et al. (2000) Immunization of humans with recombinant pneumococcal surface protein A (rPspA) elicits antibodies that passively protect mice from fatal infection with Streptococcus pneumoniae bearing heterologous PspA. J Infect Dis 182: 1694–1701. doi: 10.1086/317602 11069242

17. Ferreira DM, Darrieux M, Oliveira ML, Leite LC, Miyaji EN (2008) Optimized immune response elicited by a DNA vaccine expressing pneumococcal surface protein a is characterized by a balanced immunoglobulin G1 (IgG1)/IgG2a ratio and proinflammatory cytokine production. Clin Vaccine Immunol 15: 499–505. doi: 10.1128/CVI.00400-07 18184825

18. Ferreira DM, Oliveira ML, Moreno AT, Ho PL, Briles DE, Miyaji EN. (2010) Protection against nasal colonization with Streptococcus pneumoniae by parenteral immunization with a DNA vaccine encoding PspA (Pneumococcal surface protein A). Microb Pathog 48: 205–213. doi: 10.1016/j.micpath.2010.02.009 20206678

19. Wang S, Li Y, Shi H, Scarpellini G, Torres-Escobar A, Roland KL, et al. (2010) Immune responses to recombinant pneumococcal PsaA antigen delivered by a live attenuated Salmonella vaccine. Infect Immun 78: 3258–3271. doi: 10.1128/IAI.00176-10 20479086

20. Campos IB, Darrieux M, Ferreira DM, Miyaji EN, Silva DA, Areas AP, et al. (2008) Nasal immunization of mice with Lactobacillus casei expressing the Pneumococcal Surface Protein A: induction of antibodies, complement deposition and partial protection against Streptococcus pneumoniae challenge. Microbes Infect 10: 481–488. doi: 10.1016/j.micinf.2008.01.007 18403234

21. Hanniffy SB, Carter AT, Hitchin E, Wells JM (2007) Mucosal delivery of a pneumococcal vaccine using Lactococcus lactis affords protection against respiratory infection. J Infect Dis 195: 185–193. doi: 10.1086/509807 17191163

22. Renauld-Mongenie G, Mielcarek N, Cornette J, Schacht AM, Capron A, Riveau G, et al. (1996) Induction of mucosal immune responses against a heterologous antigen fused to filamentous hemagglutinin after intranasal immunization with recombinant Bordetella pertussis. Proc Natl Acad Sci U S A 93: 7944–7949. doi: 10.1073/pnas.93.15.7944 8755582

23. Mielcarek N, Riveau G, Remoue F, Antoine R, Capron A, Locht C. (1998) Homologous and heterologous protection after single intranasal administration of live attenuated recombinant Bordetella pertussis. Nat Biotechnol 16: 454–457. doi: 10.1038/nbt0598-454 9592394

24. Coppens I, Alonso S, Antoine R, Jacob-Dubuisson F, Renauld-Mongenie G, Jacobs E, et al. (2001) Production of Neisseria meningitidis transferrin-binding protein B by recombinant Bordetella pertussis. Infect Immun 69: 5440–5446. doi: 10.1128/IAI.69.9.5440-5446.2001 11500415

25. Li R, Lim A, Ow ST, Phoon MC, Locht C, Chow VT, et al. (2011) Development of live attenuated Bordetella pertussis strains expressing the universal influenza vaccine candidate M2e. Vaccine 29: 5502–5511. doi: 10.1016/j.vaccine.2011.05.052 21624415

26. Poulain-Godefroy O, Menozzi FD, Alonso S, Vendeville C, Capron A, Locht C, et al. (2003) Adjuvant activity of free Bordetella pertussis filamentous haemagglutinin delivered by mucosal routes. Scand J Immunol 58: 503–510. doi: 10.1046/j.1365-3083.2003.01336.x 14629622

27. Alonso S, Reveneau N, Pethe K, Locht C (2002) Eighty-kilodalton N-terminal moiety of Bordetella pertussis filamentous hemagglutinin: adherence, immunogenicity, and protective role. Infect Immun 70: 4142–4147. doi: 10.1128/IAI.70.8.4142-4147.2002 12117922

Článek vyšel v časopise


2020 Číslo 1
Nejčtenější tento týden