Engineering a New Vaccine Platform for Heterologous Antigen Delivery in Live-attenuated

Overview

Journal Comput Struct Biotechnol J

Specialty Biotechnology

Date 2021 Aug 25

PMID 34429847

Citations 4

Authors

Esther Broset

Juan Calvet Seral

Carmen Arnal

Santiago Uranga

Alex I Kanno

Luciana C C Leite

Carlos Martin

Jesus Gonzalo-Asensio

Affiliations

Soon will be listed here.

Abstract

Live vaccines are attractive vehicles for antigen delivery as a strategy to immunize against heterologous pathogens. The live vaccine MTBVAC is based on rational attenuation of with the objective of improving BCG protection against pulmonary tuberculosis. However, the development of recombinant mycobacteria as antigen-presenting microorganisms has been hindered due to their fastidious genetic manipulation. In this study, we used MTBVAC as a genetic platform to deliver diphtheria, tetanus, or pertussis toxoids, which are the immunogenic constituents of the DTP vaccine. When using nonoptimal genetic conditions, the expression of these immunogens was barely detectable. Accordingly, we pursued a rational, step-by-step optimization of the genetic components to achieve the expression and secretion of these toxoids. We explored variants of the L5 mycobacteriophage promoter to ensure balanced antigen expression and plasmid stability. Optimal signal sequences were identified by comparative proteomics of MTBVAC and its parental strain. It was determined that proteins secreted by the Twin Arginine Translocation pathway displayed higher secretion in MTBVAC, and the Ag85A secretion sequence was selected as the best candidate. Because the coding regions of diphtheria, tetanus, and pertussis toxoids significantly differ in G + C content relative to mycobacterial genes, their codon usage was optimized. We also placed a 3xFLAG epitope in frame with the C-terminus of these toxoids to facilitate protein detection. Altogether, these optimizations resulted in the secretion of DTP antigens by MTBVAC, as demonstrated by western blot and MRM-MS. Finally, we examined specific antibody responses in mice vaccinated with recombinant MTBVAC expressing DTP antigens.

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References

Netea M, Schlitzer A, Placek K, Joosten L, Schultze J . Innate and Adaptive Immune Memory: an Evolutionary Continuum in the Host's Response to Pathogens. Cell Host Microbe. 2019; 25(1):13-26. DOI: 10.1016/j.chom.2018.12.006. View

Camacho L, Ensergueix D, Perez E, Gicquel B, Guilhot C . Identification of a virulence gene cluster of Mycobacterium tuberculosis by signature-tagged transposon mutagenesis. Mol Microbiol. 1999; 34(2):257-67. DOI: 10.1046/j.1365-2958.1999.01593.x. View

Borgers K, Vandewalle K, Festjens N, Callewaert N . A guide to Mycobacterium mutagenesis. FEBS J. 2019; 286(19):3757-3774. DOI: 10.1111/febs.15041. View

Mazzantini R, Miyaji E, Dias W, Sakauchi D, Nascimento A, Raw I . Adjuvant activity of Mycobacterium bovis BCG expressing CRM197 on the immune response induced by BCG expressing tetanus toxin fragment C. Vaccine. 2004; 22(5-6):740-6. DOI: 10.1016/j.vaccine.2003.08.017. View

Broset E, Saubi N, Guitart N, Aguilo N, Uranga S, Kilpelainen A . MTBVAC-Based TB-HIV Vaccine Is Safe, Elicits HIV-T Cell Responses, and Protects against in Mice. Mol Ther Methods Clin Dev. 2019; 13:253-264. PMC: 6395831. DOI: 10.1016/j.omtm.2019.01.014. View

Martinon-Torres F, Heininger U, Thomson A, Wirsing von Konig C . Controlling pertussis: how can we do it? A focus on immunization. Expert Rev Vaccines. 2018; 17(4):289-297. DOI: 10.1080/14760584.2018.1445530. View

Kanno A, Goulart C, Rofatto H, Oliveira S, Leite L, McFadden J . New Recombinant Mycobacterium bovis BCG Expression Vectors: Improving Genetic Control over Mycobacterial Promoters. Appl Environ Microbiol. 2016; 82(8):2240-2246. PMC: 4959472. DOI: 10.1128/AEM.03677-15. View

Yeung K, Duclos P, Nelson E, Hutubessy R . An update of the global burden of pertussis in children younger than 5 years: a modelling study. Lancet Infect Dis. 2017; 17(9):974-980. DOI: 10.1016/S1473-3099(17)30390-0. View

Nascimento I, Dias W, Mazzantini R, Miyaji E, Gamberini M, Quintilio W . Recombinant Mycobacterium bovis BCG expressing pertussis toxin subunit S1 induces protection against an intracerebral challenge with live Bordetella pertussis in mice. Infect Immun. 2000; 68(9):4877-83. PMC: 101688. DOI: 10.1128/IAI.68.9.4877-4883.2000. View

10.

Marques-Neto L, Piwowarska Z, Kanno A, Moraes L, Trentini M, Rodriguez D . Thirty years of recombinant BCG: new trends for a centenary vaccine. Expert Rev Vaccines. 2021; 20(8):1001-1011. DOI: 10.1080/14760584.2021.1951243. View

11.

Tarancon R, Dominguez-Andres J, Uranga S, Ferreira A, Groh L, Domenech M . New live attenuated tuberculosis vaccine MTBVAC induces trained immunity and confers protection against experimental lethal pneumonia. PLoS Pathog. 2020; 16(4):e1008404. PMC: 7117655. DOI: 10.1371/journal.ppat.1008404. View

12.

Brosch R, Gordon S, Garnier T, Eiglmeier K, Frigui W, Valenti P . Genome plasticity of BCG and impact on vaccine efficacy. Proc Natl Acad Sci U S A. 2007; 104(13):5596-601. PMC: 1838518. DOI: 10.1073/pnas.0700869104. View

13.

Arts R, Moorlag S, Novakovic B, Li Y, Wang S, Oosting M . BCG Vaccination Protects against Experimental Viral Infection in Humans through the Induction of Cytokines Associated with Trained Immunity. Cell Host Microbe. 2018; 23(1):89-100.e5. DOI: 10.1016/j.chom.2017.12.010. View

14.

Zmora N, Zilberman-Schapira G, Suez J, Mor U, Dori-Bachash M, Bashiardes S . Personalized Gut Mucosal Colonization Resistance to Empiric Probiotics Is Associated with Unique Host and Microbiome Features. Cell. 2018; 174(6):1388-1405.e21. DOI: 10.1016/j.cell.2018.08.041. View

15.

HUNG P, MORIN J, Lubeck M, Barton J, Molnar-Kimber K, Mason B . Expression of HBV surface antigen or HIV envelope protein using recombinant adenovirus vectors. Nat Immun Cell Growth Regul. 1988; 7(3):135-43. View

16.

Gonzalo-Asensio J, Marinova D, Martin C, Aguilo N . MTBVAC: Attenuating the Human Pathogen of Tuberculosis (TB) Toward a Promising Vaccine against the TB Epidemic. Front Immunol. 2018; 8:1803. PMC: 5736532. DOI: 10.3389/fimmu.2017.01803. View

17.

Angelidou A, Diray-Arce J, Conti M, Smolen K, van Haren S, Dowling D . BCG as a Case Study for Precision Vaccine Development: Lessons From Vaccine Heterogeneity, Trained Immunity, and Immune Ontogeny. Front Microbiol. 2020; 11:332. PMC: 7078104. DOI: 10.3389/fmicb.2020.00332. View

18.

Calvo A, Moreno-Igoa M, Mancuso R, Manzano R, Olivan S, Munoz M . Lack of a synergistic effect of a non-viral ALS gene therapy based on BDNF and a TTC fusion molecule. Orphanet J Rare Dis. 2011; 6:10. PMC: 3072305. DOI: 10.1186/1750-1172-6-10. View

19.

Sato H, Ito A, Chiba J, Sato Y . Monoclonal antibody against pertussis toxin: effect on toxin activity and pertussis infections. Infect Immun. 1984; 46(2):422-8. PMC: 261549. DOI: 10.1128/iai.46.2.422-428.1984. View

20.

Tameris M, Mearns H, Penn-Nicholson A, Gregg Y, Bilek N, Mabwe S . Live-attenuated Mycobacterium tuberculosis vaccine MTBVAC versus BCG in adults and neonates: a randomised controlled, double-blind dose-escalation trial. Lancet Respir Med. 2019; 7(9):757-770. DOI: 10.1016/S2213-2600(19)30251-6. View