Abstract
It is still a challenge to develop needle-free mucosal vaccines. Despite progress in the development of the influenza vaccine, it must be reformulated annually because of antigenic changes in circulating influenza viral strains. Due to seasonal drift and shift of circulating strains, the influenza vaccine does not always match the circulating strains, and included adjuvants are not sufficient to induce a protective effect with long-lived memory cells. The adjuvants play a major role in the immune responses to a vaccine. Interestingly, the Bacillus anthracis detoxified anthrax edema toxin, which composes of protective antigen PA and N-fragment of edema factor (EFn), has shown improved effects for humoral and cellular immune responses. Here we describe the design of a universal influenza vaccine construct that consists of three tandem M2e repeats of the influenza antigen plus HA2 and detoxified toxin EFn, which is associated with the PA component, as well as the techniques used to corroborate protection. We present two major parts of description to demonstrate the vaccine strategy, using detoxified anthrax toxin for intranasal delivery of influenza antigen: (1) vaccine candidate design, production, and purification; (2) influenza virus microneutralization assay and cellular responses and lethal challenge with influenza viruses and B. anthracis Sterne spores. In the methods detailed here, we used different versions of the M2e–HA2 proteins.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Young JA, Collier RJ (2007) Anthrax toxin: receptor binding, internalization, pore formation, and translocation. Annu Rev Biochem 76:243–265
Tonello F, Montecucco CJ (2009) The anthrax lethal factor and its MAPK kinase-specific metalloprotease activity. Mol Aspects Med 30(6):431–438
Xu Q, Zeng M (2008) Detoxified lethal toxin as a potential mucosal vaccine against anthrax. Clin Vaccine Immunol 15(4):612–616
Arévalo MT, Li J, Diaz-Arévalo D, Chen Y, Navarro A, Wu L et al (2017) A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax. Immunology 150(3):276–289
Meng Q, Liu G, Liu Y, Deng X, Wang W, Xu K et al (2015) A broad protection provided by matrix protein 2 (M2) of avian influenza virus. Vaccine 33(31):3758–3765
Chen J, Lee KH, Steinhauer DA, Stevens DJ, Skehel JJ, Wiley DC (1998) Structure of the hemagglutinin precursor cleavage site, a determinant of influenza pathogenicity and the origin of the labile conformation. Cell 95(3):409–417
Edwards M, Dimmock NJ (2000) Two influenza A virus-specific Fabs neutralize by inhibiting virus attachment to target cells, while neutralization by their IgGs is complex and occurs simultaneously through fusion inhibition and attachment inhibition. Virology 278(2):423–435
Throsby M, van den Brink E, Jongeneelen M, Poon LL, Alard P, Cornelissen L et al (2008) Heterosubtypic neutralizing monoclonal antibodies cross-protective against H5N1 and H1N1 recovered from human IgM+ memory B cells. PLoS One 3(12):e3942
Wang TT, Tan GS, Hai R, Pica N, Ngai L, Ekiert DC et al (2010) Vaccination with a synthetic peptide from the influenza virus hemagglutinin provides protection against distinct viral subtypes. Proc Natl Acad Sci U S A 107(44):18979–18984
Turnbull P, Broster MG, Carman JA, Manchee RJ, Melling J (1986) Development of antibodies to protective antigen and lethal factor components of anthrax toxin in humans and guinea pigs and their relevance to protective immunity. Infect Immun 52(2):356–363
Ingram RJ, Metan G, Maillere B, Doganay M, Ozkul Y, Kim LU et al (2010) Natural exposure to cutaneous anthrax gives long-lasting T cell immunity encompassing infection-specific epitopes. J Immunol 184(7):3814–3821
Quesnel-Hellmann A, Cleret A, Vidal DR, Tournier JN (2006) Evidence for adjuvanticity of anthrax edema toxin. Vaccine 24(6):699–702
Duverger A, Jackson RJ, Van Ginkel FW, Fischer R, Tafaro A, Leppla SH et al (2006) Bacillus anthracis edema toxin acts as an adjuvant for mucosal immune responses to nasally administered vaccine antigens. J Immunol 176(3):1776–1783
Zeng M, Xu Q, Hesek ED, Pichichero ME (2006) N-fragment of edema factor as a candidate antigen for immunization against anthrax. Vaccine 24(5):662–670
Li J, Arévalo MT, Chen Y, Posadas O, Smith JA, Zeng M (2014) Intranasal immunization with influenza antigens conjugated with cholera toxin subunit B stimulates broad spectrum immunity against influenza viruses. Hum Vaccin Immunother 10(5):1211–1220
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Diaz-Arévalo, D., Chen, Y., Zeng, M. (2021). Vaccine Delivery with a Detoxified Bacterial Toxin. In: Pfeifer, B.A., Hill, A. (eds) Vaccine Delivery Technology. Methods in Molecular Biology, vol 2183. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0795-4_22
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0795-4_22
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0794-7
Online ISBN: 978-1-0716-0795-4
eBook Packages: Springer Protocols