Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses

Document Type : Original Article

Authors

1 Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of Immunology, Pasteur Institute of Iran, 69 Pasteur Ave., Tehran, Iran

Abstract

Objective(s): After decades of containment, pertussis disease, caused by Bordetella pertussis seems to be re-emerging and still remains a major cause of reported vaccine-preventable deaths worldwide. The current licensed whole-cell vaccines display reactogenicity while acellular vaccines are expensive and do not induce Th1-type immune responses that are required for optimum protection against the disease. Thus, there is an urgent need to develop new vaccines and the recombinant technology seems to be the method of choice for this purpose. The present study was an attempt to develop a new, simplified, cost-effective and well-defined vaccine against Bordetella pertussis, with capacity to induce a Th1 response.
Materials and Methods: A fusion DNA fragment encoding the N-terminal region of pertussis toxin S1 subunit and filamentous hemagglutinin type 1 immunodominant domain was constructed and the corresponding fusion protein (F1S1) was produced in Escherichia coli. F1S1 in conjunction with imiquimod was administered by subcutaneous (SC) and intranasal (IN) routes to BALB/c mice.
Results: This vaccine formulation could elicit high levels of IFN-γ, serum IgG (with higher IgG2a/IgG1 ratio) and lung IgA after the SC and, to a lesser extent, following the IN administration.  
Conclusion: Our results indicate that the above-mentioned important proteins of B. pertussis could be successfully produced in E. coli as a single fusion protein. Furthermore, this protein could induce proper systemic and mucosal immune responses after administration via SC or IN routes.

Keywords

Main Subjects


1.    Ulloa-Gutierrez R, Boza R, Carvajal-Riggioni D, Baltodano A. Pertussis: should we improve intensive care management or vaccination strategies? Expert Rev Vaccines. 2011;10:49-53.
2.    Melvin JA, Scheller EV, Miller JF, Cotter PA. Bordetella pertussis pathogenesis: current and future challenges. Nat Rev Microbiol. 2014;12:274-288.
3.    Dorji D, Mooi F, Yantorno O, Deora R, Graham RM, Mukkur TK. Bordetella pertussis virulence factors in the continuing evolution of whooping cough vaccines for improved performance. Med Microbiol Immunol. 2018;207:3-26.
4.    Berbers GA, de Greeff SC, Mooi FR. Improving pertussis vaccination. Hum Vaccin. 2009; 5:497-503.
5.    Mahon BP, Brady MT, Mills KH. Protection against Bordetella pertussis in mice in the absence of detectable circulating antibody: implications for long-term immunity in children. J Infect Dis. 2000;181:2087-2091.
6.    van der Lee S, Hendrikx LH, Sanders EAM, Berbers GAM, Buisman AM. Whole-cell or acellular pertussis primary immunizations in infancy determines adolescent cellular immune profiles. Front Immunol. 2018;9:51.
7.    Higgs R, Higgins SC, Ross PJ, Mills KH. Immunity to the respiratory pathogen Bordetella pertussis. Mucosal Immunol. 2012;5:485-500.
8.    Allen AC, Mills KH. Improved pertussis vaccines based on adjuvants that induce cell-mediated immunity. Expert Rev Vaccines. 2014;13:1253-1264.
9.    Schure RM, Hendrikx LH, de Rond LG, Ozturk K, Sanders EA, Berbers GA, et al. T-cell responses before and after the fifth consecutive acellular pertussis vaccination in 4-year-old Dutch children. Clin Vaccine Immunol. 2012;19:1879-1886.
10.    Ross PJ, Sutton CE, Higgins S, Allen AC, Walsh K, Misiak A, et al. Relative contribution of Th1 and Th17 cells in adaptive immunity to Bordetella pertussis: towards the rational design of an improved acellular pertussis vaccine. PLoS Pathog. 2013;9:e1003264.
11.    Misiak A, Leuzzi R, Allen AC, Galletti B, Baudner BC, D’Oro U, et al. Addition of a TLR7 agonist to an acellular pertussis vaccine enhances Th1 and Th17 responses and protective immunity in a mouse model. Vaccine. 2017;35:5256-5263.
12.    Latasa P, Garcia-Comas L, Gil de Miguel A, Barranco MD, Rodero I, Sanz JC, et al. Effectiveness of acellular pertussis vaccine and evolution of pertussis incidence in the community of Madrid from 1998 to 2015. Vaccine. 2018;36:1643-1649.
13.    Seubert A, D’Oro U, Scarselli M, Pizza M. Genetically detoxified pertussis toxin (PT-9K/129G): implications for immunization and vaccines. Expert Rev Vaccines. 2014;13:1191-1204.
14.    Ochiai M, Horiuchi Y, Yuen CT, Asokanathan C, Yamamoto A, Okada K, et al. Investigation in a murine model of possible mechanisms of enhanced local reactions to post-primary diphtheria-tetanus toxoid boosters in recipients of acellular pertussis-diphtheria-tetanus vaccine. Hum Vaccin Immunother. 2014;10:2074-2080.
15.    Kamachi K, Arakawa Y. Expression of a C terminally truncated form of pertussis toxin S1 subunit effectively induces protection against pertussis toxin following DNA-based immunization. Infect Immun. 2004;72:4293-4296.
16.    Kamachi K, Konda T, Arakawa Y. DNA vaccine encoding pertussis toxin S1 subunit induces protection against Bordetella pertussis in mice. Vaccine. 2003;21:4609-4615.
17.    Scheller EV, Cotter PA. Bordetella filamentous hemagglutinin and fimbriae: critical adhesins with unrealized vaccine potential. Pathog Dis. 2015;73:ftv079.
18.    Knight JB, Huang YY, Halperin SA, Anderson R, Morris A, Macmillan A, et al. Immunogenicity and protective efficacy of a recombinant filamentous haemagglutinin from Bordetella pertussis. Clin Exp Immunol. 2006;144:543-551.
19.    Cherry JD. Comparative efficacy of acellular pertussis vaccines: an analysis of recent trials. Pediatr Infect Dis J. 1997;16:S90-S96.
20.    Delisse-Gathoye AM, Locht C, Jacob F, Raaschou-Nielsen M, Heron I, Ruelle JL, et al. Cloning, partial sequence, expression, and antigenic analysis of the filamentous hemagglutinin gene of Bordetella pertussis. Infect Immun. 1990;58:2895-2905.
21.    Raman VS, Duthie MS, Fox CB, Matlashewski G, Reed SG. Adjuvants for Leishmania vaccines: from models to clinical application. Front Immunol. 2012;3:144.
22.    Jinyong Z, Xiaoli Z, Weijun Z, Ying G, Gang G, Xuhu M, et al. Fusion expression and immunogenicity of Bordetella pertussis PTS1-FHA protein: implications for the vaccine development. Mol Biol Rep. 2011 Mar;38(3):1957-63.
23.    Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-254.
24.    Reynolds E, Walker B, Xing D, Southern J, Asokanathan C, Dagg B, et al. Laboratory investigation of immune responses to acellular pertussis vaccines when used for boosting adolescents after primary immunisation with whole cell pertussis vaccines: a comparison with data from clinical study. Vaccine. 2006;24:3248-3257.
25.    Hung IF, Zhang AJ, To KK, Chan JF, Li C, Zhu HS, et al. Immunogenicity of intradermal trivalent influenza vaccine with topical imiquimod: a double blind randomized controlled trial. Clin Infect Dis. 2014;59:1246-1255.
26.    Ryan M, Gothefors L, Storsaeter J, Mills KH. Bordetella pertussis-specific Th1/Th2 cells generated following respiratory infection or immunization with an acellular vaccine: comparison of the T cell cytokine profiles in infants and mice. Dev Biol Stand. 1997;89:297-305.
27.    He P, Zou Y, Hu Z. Advances in aluminum hydroxide-based adjuvant research and its mechanism. Hum Vaccin Immunother. 2015;11:477-488.
28.    Aimanianda V, Haensler J, Lacroix-Desmazes S, Kaveri SV, Bayry J. Novel cellular and molecular mechanisms of induction of immune responses by aluminum adjuvants. Trends Pharmacol Sci. 2009; 30:287-295.
29.    Jurk M, Heil F, Vollmer J, Schetter C, Krieg AM, Wagner H, et al. Human TLR7 or TLR8 independently confer responsiveness to the antiviral compound R-848. Nat Immunol. 2002;3:499.
30.    Mills KH. Immunity to Bordetella pertussis. Microbes Infect. 2001;3:655-677.
31.    Hellwig SM, van Spriel AB, Schellekens JF, Mooi FR, van de Winkel JG. Immunoglobulin A-mediated protection against Bordetella pertussis infection. Infect Immun. 2001;69:4846-4850.
32.    Clements JD, Freytag LC. Parenteral vaccination can be an effective means of inducing protective mucosal responses. Clin Vaccine Immunol. 2016;23:438-441.