CbpM and CbpG of Streptococcus Pneumoniae Elicit a High Protection in Mice Challenged with a Serotype 19F Pneumococcus

  • Hamid Kazemian Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Davoud Afshar Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Ernesto Garcia Centro de Investigación Biomédica en Red de Enfermedades Respiratorias, Madrid, Spain
  • Mohammad Reza Pourmand Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AND Urology Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran
  • Farzaneh Aminharati Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Fazel Shokri Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Hossein Yazdi Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran AND Biotechnology Research Center, Tehran University of Medical Sciences, Tehran, Iran
Keywords: Choline-binding proteins, Pneumococcal vaccine, Streptococcus pneumoniae

Abstract

Among many pneumococcal antigens, choline-binding proteins (CPBs) display a high immunogenicity in animal models. This study aims to determine the immunogenicity of CbpM, CbpG and CbpL proteins of Streptococcus pneumoniae in a mice model. The genes were cloned into pET21a expression vector and the recombinant proteins were produced. Mice were immunized with the purified recombinant proteins. Subsequently, the mice were challenged with S. pneumoniae ATCC 49619 (2×106 CFU) and their survival and bacterial clearances were followed 24 hours after infection. The antibody responses of the mice were determined by ELISA assay. The opsonophagocytosis assay was performed using rabbit’s sera. Passive immunization was carried out using two doses of anti-CbPs antibodies. Finally, these mice were  experimentally infected with virulent bacteria and the protective effects of two doses of 10 and 100 µg/mL by monitoring the survival rate and bacterial clearance were determined at 2, 3 and 7 days after bacterial challenge. The mice actively immunized with CbpM, CbpG and CbpL recombinant proteins showed survival rate of 100%, 85% and 75%, respectively. The survival rates among passively immunized mice groups which received 100 µg/mL dose of anti-CbpM, anti-CbpG and anti- CbpL were 50%, 50% and 25%, respectively. The rates of opsonization with rabbit’s antibodies against CbpM, CbpG, and CbpL  at 100 µg/mL doses was 45.6%, 14.7% and 82.3%, and at 10 µg/mL was 12.9%, 12.2% and 9.35%, respectively. Our findings suggest that the recombinant proteins particularly CbpM and CbpG can protect the mice against pneumococcus19F serotype and effectively induce a protective antibody response. Thus, CbpG and CbpM proteins might be used as suitable vaccine candidate in pneumococcal vaccine formulations.

Author Biographies

Hamid Kazemian, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Department of Pathobiology, School of Public Health

Davoud Afshar, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Department of Pathobiology, School of Public Health

Ernesto Garcia, Centro de Investigación Biomédica en Red de Enfermedades Respiratorias, Madrid, Spain

Centro de Investigación Biomédica

Mohammad Reza Pourmand, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AND Urology Research Center, Tehran University of Medical Sciences, Tehran, Iran

Department of Pathobiology, School of Public Health

Mahmood Jeddi-Tehrani, Monoclonal Antibody Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran

Monoclonal Antibody Research Center,

Farzaneh Aminharati, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Department of Pathobiology, School of Public Health

Fazel Shokri, Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Department of Immunology, School of Public Health,

References

1. Afshar D, Pourmand MR, Jeddi-Tehrani M, Saboor Yaraghi AA, Azarsa M, Shokri F. Fibrinogen and Fibronectin Binding Activity and Immunogenic Nature of Choline Binding Protein M. Iranian Journal of Public Health 2016; 45(12):1610-1617.

2. Arulanandam BP, Lynch JM, brile de, hollingshead s, metzgar dw. Intranasal vaccination with pneumococcal surface protein A and interleukin-12 augments antibody-mediated opsonization and protective immunity against Streptococcus pneumoniae infection. Infection and immunity 2001; 69(11):6718-6724.

3. Balachandran P, Brooks-Walter A, Virolainen-Julkunen A, Hollingshead SK, Briles DE.. Role of pneumococcal surface protein C in nasopharyngeal carriage and pneumonia and its ability to elicit protection against carriage of Streptococcus pneumoniae. Infection and immunity 2002; 70(5):2526-2534.

4. Brown J, Hammerschmidt S, Orihuela C. Streptococcus Pneumoniae: Molecular Mechanisms of Host-Pathogen Interactions. Academic Press, 2015.

5. Chen A, Mann B, Gao G, Heath R, King J, Maissoneuve J, et al. Multivalent pneumococcal protein vaccines comprising pneumolysoid with epitopes/fragments of CbpA and/or PspA elicit strong and broad protection. Clinical and Vaccine Immunology 2015; 22(10):1079-1089.

6. Corsini B, Aguinagalde L, Ruiz S, Domenech M, Antequera ML, Fenoll A, et al. Immunization with LytB protein of Streptococcus pneumoniae activates complement-mediated phagocytosis and induces protection against pneumonia and sepsis. Vaccine 2016; 34(50):6148-6157.

7. Darrieux M, Goulart C, Briles D, Leite LC.. Current status and perspectives on protein-based pneumococcal vaccines. Critical reviews in microbiology 2015; 41(2):190-200.

8. Frolet C, Beniazza M, Roux L, Gallet B, Noirclerc-Savoye M, Vernet T, et al. New adhesin functions of surface-exposed pneumococcal proteins. BMC Microbiol 2010;10:190.

9. Gosink KK, Mann ER, Guglielmo C, Tuomanen EI, Masure HR. Role of novel choline binding proteins in virulence of Streptococcus pneumoniae. Infect Immun 2000; 68(10):5690-5695.

10. Grabenstein J, Klugman K. A century of pneumococcal vaccination research in humans. Clinical Microbiology and Infection 2012; 18(5):15-24.

11. Gruber WC, Scott DA, Emini EA. Development and clinical evaluation of Prevnar 13, a 13‐valent pneumocococcal CRM197 conjugate vaccine. Annals of the New York Academy of Sciences 2012; 1263(1):15-26.

12. Guo B, Zhao X, Shi Y. Pathogenic implication of a fibrinogen-binding protein of Staphylococcus epidermidis in a rat model of intravascular-catheter-associated infection. Infection and immunity 2007; 75(6):2991-2995.

13. Gutierrez-Fernandez J, Saleh M, Alcorlo M, Gómez-Mejía A, Pantoja-Uceda D, Treviño MA, et al. Modular Architecture and Unique Teichoic Acid Recognition Features of Choline-Binding Protein L (CbpL) Contributing to Pneumococcal Pathogenesis. Sci Rep 2016;6:38094.

14. Hanage WP, Finkelstein JA, Huang SS, Pelton SI, Stevenson AE, Kleinman K, et al. Evidence that pneumococcal serotype replacement in Massachusetts following conjugate vaccination is now complete. Epidemics 2010; 2(2):80-84.

15. Hausdorff WP, Brueggemann AB, Hackell JG, Scott AJ. Pneumococcal serotype epidemiology 2008; Washington, DC: ASM Press.

16. Jedrzejas MJ. Pneumococcal virulence factors: structure and function. Microbiology and Molecular Biology Reviews 2001; 65(2):187-207.

17. Kadioglu A, Weiser JN, Paton JC, Andrew PW. The role of Streptococcus pneumoniae virulence factors in host respiratory colonization and disease. Nature Reviews Microbiology 2008; 6(4):288-301.

18. Keller LE, Luo X, Thornton JA, Seo KS, Moon BY, Robinson DA, et al. Immunization with pneumococcal surface protein K of nonencapsulated Streptococcus pneumoniae provides protection in a mouse model of colonization. Clinical and Vaccine Immunology 2015; 22(11):1146-1153.

19. Lipsitch M. Bacterial vaccines and serotype replacement: lessons from Haemophilus influenzae and prospects for Streptococcus pneumoniae. Emerging infectious diseases 1999; 5(3):336.

20. Maestro B, Sanz JM. Choline Binding Proteins from Streptococcus pneumoniae: A Dual Role as Enzybiotics and Targets for the Design of New Antimicrobials. Antibiotics 2016; 5(2):21.

21. Mangtani P, Cutts F, Hall AJ. Efficacy of polysaccharide pneumococcal vaccine in adults in more developed countries: the state of the evidence. The Lancet infectious diseases 2003; 3(2):71-78.

22. Ogunniyi AD, Grabowicz M, Briles DE, Cook J, Paton JC. Development of a vaccine against invasive pneumococcal disease based on combinations of virulence proteins of Streptococcus pneumoniae. Infection and immunity 2007; 75(1):350-357.

23. Ogunniyi AD, Woodrow MC, Poolman JT, Paton JC. Protection against Streptococcus pneumoniae elicited by immunization with pneumolysin and CbpA. Infection and immunity 2001; 69(10):5997-6003.

24. Park S, Nahm MH. Older adults have a low capacity to opsonize pneumococci due to low IgM antibody response to pneumococcal vaccinations. Infection and immunity 2011; 79(1):314-20.

25. Swiatlo E, Champlin FR, Holman SC, Wilson WW, Watt JM. Contribution of choline-binding proteins to cell surface properties of Streptococcus pneumoniae. Infection and immunity 2002; 70(1):412-5.

26. Weinberger DM, Malley R, Lipsitch M. Serotype replacement in disease after pneumococcal vaccination. The Lancet 2011; 378(9807):1962-73.

Published
2018-12-03
How to Cite
1.
Kazemian H, Afshar D, Garcia E, Pourmand MR, Jeddi-Tehrani M, Aminharati F, Shokri F, Yazdi MH. CbpM and CbpG of Streptococcus Pneumoniae Elicit a High Protection in Mice Challenged with a Serotype 19F Pneumococcus. Iran J Allergy Asthma Immunol. 17(6):574-585.
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Original Article(s)