MTBVAC: Attenuating the Human Pathogen of Tuberculosis (TB) Toward a Promising Vaccine against the TB Epidemic

Jesus Gonzalo-Asensio, Dessislava Marinova, Carlos Martin, Nacho Aguilo, Jesus Gonzalo-Asensio, Dessislava Marinova, Carlos Martin, Nacho Aguilo

Abstract

Bacille Calmette-Guérin (BCG) is a live-attenuated strain of Mycobacterium bovis developed a century ago by repeated subculture. It remains the only vaccine against tuberculosis (TB) in use today, and it offers variable protection against the respiratory forms of TB responsible for transmission. The principal genetic basis for BCG attenuation is the loss of the region of difference 1 (RD1) that includes the genes codifying for production and export of the major virulence factor ESAT6. Today more than 13 TB vaccine candidates are in clinical evaluation. One of these candidates is MTBVAC, which is based on a rationally attenuated Mycobacterium tuberculosis clinical isolate belonging to modern lineage 4, one of the most widespread lineages among humans. MTBVAC conserves most of the T cell epitopes described for TB including the major immunodominant antigens ESAT6 and CFP10 of the RD1, deleted in BCG. After almost 20 years of discovery and preclinical development, MTBVAC is the only live attenuated vaccine based on a human pathogen that has successfully entered clinical trials as a preventive vaccine in newborns, aiming to replace BCG, and as a preventive vaccine in adolescents and adults (BCG-vaccinated at birth). Our recent preclinical studies have demonstrated that MTBVAC-induced immunity to ESAT6 and CFP10 correlate with improved efficacy relative to BCG encouraging exploration of these responses in human clinical trials as potential biomarkers and identification of these antigens as possible correlates of vaccine-induced protection. Such data would be extremely valuable as they would greatly accelerate clinical development to efficacy trials.

Keywords: Bacille Calmette-Guérin; CFP10; ESAT6; MTBVAC; live vaccines; tuberculosis.

Figures

Figure 1
Figure 1
Genomic deletions between human and cattle tuberculosis pathogens. Eight regions of difference (RD) deleted in the bovine pathogen Mycobacterium bovis with respect to the human pathogen Mycobacterium tuberculosis. Repeated subcultivation of an M. bovis strain for 13 years (1908–1921), following classical Pasteur’s postulates, led to attenuation due to loss of RD1, giving rise to Bacille Calmette-Guérin (BCG). MTBVAC is the result of the rational attenuation of an M. tuberculosis clinical isolate by genetic deletions of the two independent virulence genes phoP and fadD26, following molecular Pasteur’s postulates for attenuated vaccines.
Figure 2
Figure 2
Localization of 1,603 experimentally demonstrated T cell epitopes (according to http://IEDB.org) in the Mycobacterium tuberculosis H37Rv chromosome. Those regions absent in Bacille Calmette-Guérin (BCG) but present in M. tuberculosis are shown by red sectors and the percentage of epitopes contained in each region is indicated. Three of these regions (RD1, RD2, and RD11) contain >25% of the total M. tuberculosis epitopes. It is also important to note that Ag85B (shown by a yellow sector) produced but not secreted by BCG also contains a high percentage of epitopes. Taken together, these results indicate that live attenuated vaccines based on the human pathogen, as MTBVAC, would present almost 40% more T cell epitopes than BCG.

References

    1. Brosch R, Gordon SV, Garnier T, Eiglmeier K, Frigui W, Valenti P, et al. Genome plasticity of BCG and impact on vaccine efficacy. Proc Natl Acad Sci U S A (2007) 104:5596–601.10.1073/pnas.0700869104
    1. Kaufmann SH, Weiner J, Reyn CF. Novel approaches to TB vaccine development. Int J Infect Dis (2016) 56:263–67.10.1016/j.ijid.2016.10.018
    1. Colditz GA, Berkey CS, Mosteller F, Brewer TF, Wilson ME, Burdick E, et al. The efficacy of bacillus Calmette-Guerin vaccination of newborns and infants in the prevention of tuberculosis: meta-analyses of the published literature. Pediatrics (1995) 96:29–35.
    1. Trunz BB, Fine P, Dye C. Effect of BCG vaccination on childhood tuberculous meningitis and miliary tuberculosis worldwide: a meta-analysis and assessment of cost-effectiveness. Lancet (2006) 367:1173–80.10.1016/S0140-6736(06)68507-3
    1. Abubakar I, Pimpin L, Ariti C, Beynon R, Mangtani P, Sterne JA, et al. Systematic review and meta-analysis of the current evidence on the duration of protection by bacillus Calmette-Guerin vaccination against tuberculosis. Health Technol Assess (2013) 17:1–372, v–vi.10.3310/hta17370
    1. WHO. Global Tuberculosis Report (2017). Available from:
    1. Ai JW, Ruan QL, Liu QH, Zhang WH. Updates on the risk factors for latent tuberculosis reactivation and their managements. Emerg Microbes Infect (2016) 5:e10.10.1038/emi.2016.10
    1. Kaufmann SH, Cotton MF, Eisele B, Gengenbacher M, Grode L, Hesseling AC, et al. The BCG replacement vaccine VPM1002: from drawing board to clinical trial. Expert Rev Vaccines (2014) 13:619–30.10.1586/14760584.2014.905746
    1. Scriba TJ, Kaufmann SH, Henri Lambert P, Sanicas M, Martin C, Neyrolles O. Vaccination against tuberculosis with whole-cell mycobacterial vaccines. J Infect Dis (2016) 214:659–64.10.1093/infdis/jiw228
    1. Arbues A, Aguilo JI, Gonzalo-Asensio J, Marinova D, Uranga S, Puentes E, et al. Construction, characterization and preclinical evaluation of MTBVAC, the first live-attenuated M. tuberculosis-based vaccine to enter clinical trials. Vaccine (2013) 31:4867–73.10.1016/j.vaccine.2013.07.051
    1. Stucki D, Brites D, Jeljeli L, Coscolla M, Liu Q, Trauner A, et al. Mycobacterium tuberculosis lineage 4 comprises globally distributed and geographically restricted sublineages. Nat Genet (2016) 48:1535–43.10.1038/ng.3704
    1. Kamath AT, Fruth U, Brennan MJ, Dobbelaer R, Hubrechts P, Ho MM, et al. New live mycobacterial vaccines: the Geneva consensus on essential steps towards clinical development. Vaccine (2005) 23:3753–61.10.1016/j.vaccine.2005.03.001
    1. Walker KB, Brennan MJ, Ho MM, Eskola J, Thiry G, Sadoff J, et al. The second Geneva consensus: recommendations for novel live TB vaccines. Vaccine (2010) 28:2259–70.10.1016/j.vaccine.2009.12.083
    1. Cox JS, Chen B, Mcneil M, Jacobs WR, Jr. Complex lipid determines tissue-specific replication of Mycobacterium tuberculosis in mice. Nature (1999) 402:79–83.10.1038/47042
    1. Trivedi OA, Arora P, Vats A, Ansari MZ, Tickoo R, Sridharan V, et al. Dissecting the mechanism and assembly of a complex virulence mycobacterial lipid. Mol Cell (2005) 17:631–43.10.1016/j.molcel.2005.02.009
    1. Kirksey MA, Tischler AD, Simeone R, Hisert KB, Uplekar S, Guilhot C, et al. Spontaneous phthiocerol dimycocerosate-deficient variants of Mycobacterium tuberculosis are susceptible to gamma interferon-mediated immunity. Infect Immun (2011) 79:2829–38.10.1128/IAI.00097-11
    1. Gonzalo-Asensio JÁ. Deciphering the Role of PhoP in Mycobacterium tuberculosis Virulence. Doctor en Bioquimica, Universidad de Zaragoza; (2006).
    1. Gonzalo-Asensio J, Mostowy S, Harders-Westerveen J, Huygen K, Hernandez-Pando R, Thole J, et al. PhoP: a missing piece in the intricate puzzle of Mycobacterium tuberculosis virulence. PLoS One (2008) 3:e3496.10.1371/journal.pone.0003496
    1. Solans L, Aguilo N, Samper S, Pawlik A, Frigui W, Martin C, et al. A specific polymorphism in Mycobacterium tuberculosis H37Rv causes differential ESAT6 expression and identifies WhiB6 as a novel ESX-1 component. Infect Immun (2014) 82:3446–56.10.1128/IAI.01824-14
    1. Solans L, Gonzalo-Asensio J, Sala C, Benjak A, Uplekar S, Rougemont J, et al. The PhoP-dependent ncRNA Mcr7 modulates the TAT secretion system in Mycobacterium tuberculosis. PLoS Pathog (2014) 10:e1004183.10.1371/journal.ppat.1004183
    1. Gonzalo Asensio J, Maia C, Ferrer NL, Barilone N, Laval F, Soto CY, et al. The virulence-associated two-component PhoP-PhoR system controls the biosynthesis of polyketide-derived lipids in Mycobacterium tuberculosis. J Biol Chem (2006) 281:1313–6.10.1074/jbc.C500388200
    1. Chesne-Seck ML, Barilone N, Boudou F, Gonzalo Asensio J, Kolattukudy PE, Martin C, et al. A point mutation in the two-component regulator PhoP-PhoR accounts for the absence of polyketide-derived acyltrehaloses but not that of phthiocerol dimycocerosates in Mycobacterium tuberculosis H37Ra. J Bacteriol (2008) 190:1329–34.10.1128/JB.01465-07
    1. Frigui W, Bottai D, Majlessi L, Monot M, Josselin E, Brodin P, et al. Control of M. tuberculosis ESAT6 secretion and specific T cell recognition by PhoP. PLoS Pathog (2008) 4:e33.10.1371/journal.ppat.0040033
    1. Martin C, Williams A, Hernandez-Pando R, Cardona PJ, Gormley E, Bordat Y, et al. The live Mycobacterium tuberculosis phoP mutant strain is more attenuated than BCG and confers protective immunity against tuberculosis in mice and guinea pigs. Vaccine (2006) 24:3408–19.10.1016/j.vaccine.2006.03.017
    1. Aguilo N, Uranga S, Marinova D, Monzon M, Badiola J, Martin C. MTBVAC vaccine is safe, immunogenic and confers protective efficacy against Mycobacterium tuberculosis in newborn mice. Tuberculosis (Edinb) (2016) 96:71–4.10.1016/j.tube.2015.10.010
    1. Aguilo N, Gonzalo-Asensio J, Alvarez-Arguedas S, Marinova D, Gomez AB, Uranga S, et al. Reactogenicity to major tuberculosis antigens absent in BCG is linked to improved protection against Mycobacterium tuberculosis. Nat Commun (2017) 8:16085.10.1038/ncomms16085
    1. Clark S, Lanni F, Marinova D, Rayner E, Martin C, Williams A. Revaccination of guinea pigs with the live attenuated Mycobacterium tuberculosis vaccine MTBVAC improves BCG’s protection against tuberculosis. J Infect Dis (2017) 216(5):525–33.10.1093/infdis/jix030
    1. Marinova D, Gonzalo-Asensio J, Aguilo N, Martin C. MTBVAC from discovery to clinical trials in tuberculosis-endemic countries. Expert Rev Vaccines (2017) 16:565–76.10.1080/14760584.2017.1324303
    1. Copin R, Coscolla M, Efstathiadis E, Gagneux S, Ernst JD. Impact of in vitro evolution on antigenic diversity of Mycobacterium bovis bacillus Calmette-Guerin (BCG). Vaccine (2014) 32:5998–6004.10.1016/j.vaccine.2014.07.113
    1. Pym AS, Brodin P, Brosch R, Huerre M, Cole ST. Loss of RD1 contributed to the attenuation of the live tuberculosis vaccines Mycobacterium bovis BCG and Mycobacterium microti. Mol Microbiol (2002) 46:709–17.10.1046/j.1365-2958.2002.03237.x
    1. Lewis KN, Liao R, Guinn KM, Hickey MJ, Smith S, Behr MA, et al. Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette-Guerin attenuation. J Infect Dis (2003) 187:117–23.10.1086/345862
    1. Romagnoli A, Etna MP, Giacomini E, Pardini M, Remoli ME, Corazzari M, et al. ESX-1 dependent impairment of autophagic flux by Mycobacterium tuberculosis in human dendritic cells. Autophagy (2012) 8:1357–70.10.4161/auto.20881
    1. Gutierrez MG, Master SS, Singh SB, Taylor GA, Colombo MI, Deretic V. Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuberculosis survival in infected macrophages. Cell (2004) 119:753–66.10.1016/j.cell.2004.11.038
    1. Dallenga T, Repnik U, Corleis B, Eich J, Reimer R, Griffiths GW, et al. M. tuberculosis-induced necrosis of infected neutrophils promotes bacterial growth following phagocytosis by macrophages. Cell Host Microbe (2017) 22:519–30.e513.10.1016/j.chom.2017.09.003
    1. Derrick SC, Morris SL. The ESAT6 protein of Mycobacterium tuberculosis induces apoptosis of macrophages by activating caspase expression. Cell Microbiol (2007) 9:1547–55.10.1111/j.1462-5822.2007.00892.x
    1. Aguilo JI, Alonso H, Uranga S, Marinova D, Arbues A, De Martino A, et al. ESX-1-induced apoptosis is involved in cell-to-cell spread of Mycobacterium tuberculosis. Cell Microbiol (2013) 15:1994–2005.10.1111/cmi.12169
    1. Davis JM, Ramakrishnan L. The role of the granuloma in expansion and dissemination of early tuberculous infection. Cell (2009) 136:37–49.10.1016/j.cell.2008.11.014
    1. Aguilo N, Marinova D, Martin C, Pardo J. ESX-1-induced apoptosis during mycobacterial infection: to be or not to be, that is the question. Front Cell Infect Microbiol (2013) 3:88.10.3389/fcimb.2013.00088
    1. Smith J, Manoranjan J, Pan M, Bohsali A, Xu J, Liu J, et al. Evidence for pore formation in host cell membranes by ESX-1-secreted ESAT6 and its role in Mycobacterium marinum escape from the vacuole. Infect Immun (2008) 76:5478–87.10.1128/IAI.00614-08
    1. Simeone R, Bobard A, Lippmann J, Bitter W, Majlessi L, Brosch R, et al. Phagosomal rupture by Mycobacterium tuberculosis results in toxicity and host cell death. PLoS Pathog (2012) 8:e1002507.10.1371/journal.ppat.1002507
    1. Simeone R, Sayes F, Song O, Groschel MI, Brodin P, Brosch R, et al. Cytosolic access of Mycobacterium tuberculosis: critical impact of phagosomal acidification control and demonstration of occurrence in vivo. PLoS Pathog (2015) 11:e1004650.10.1371/journal.ppat.1004650
    1. Wassermann R, Gulen MF, Sala C, Perin SG, Lou Y, Rybniker J, et al. Mycobacterium tuberculosis differentially activates cGAS- and inflammasome-dependent intracellular immune responses through ESX-1. Cell Host Microbe (2015) 17:799–810.10.1016/j.chom.2015.05.003
    1. McNab F, Mayer-Barber K, Sher A, Wack A, O’Garra A. Type I interferons in infectious disease. Nat Rev Immunol (2015) 15:87–103.10.1038/nri3787
    1. Aagaard C, Hoang T, Dietrich J, Cardona PJ, Izzo A, Dolganov G, et al. A multistage tuberculosis vaccine that confers efficient protection before and after exposure. Nat Med (2011) 17:189–94.10.1038/nm.2285
    1. Rogerson BJ, Jung YJ, Lacourse R, Ryan L, Enright N, North RJ. Expression levels of Mycobacterium tuberculosis antigen-encoding genes versus production levels of antigen-specific T cells during stationary level lung infection in mice. Immunology (2006) 118:195–201.10.1111/j.1365-2567.2006.02355.x
    1. Bold TD, Banaei N, Wolf AJ, Ernst JD. Suboptimal activation of antigen-specific CD4+ effector cells enables persistence of M. tuberculosis in vivo. PLoS Pathog (2011) 7:e1002063.10.1371/journal.ppat.1002063
    1. Moguche AO, Musvosvi M, Penn-Nicholson A, Plumlee CR, Mearns H, Geldenhuys H, et al. Antigen availability shapes T cell differentiation and function during tuberculosis. Cell Host Microbe (2017) 21:695–706.e695.10.1016/j.chom.2017.05.012
    1. Pym AS, Brodin P, Majlessi L, Brosch R, Demangel C, Williams A, et al. Recombinant BCG exporting ESAT6 confers enhanced protection against tuberculosis. Nat Med (2003) 9:533–9.10.1038/nm859
    1. Groschel MI, Sayes F, Shin SJ, Frigui W, Pawlik A, Orgeur M, et al. Recombinant BCG expressing ESX-1 of Mycobacterium marinum combines low virulence with cytosolic immune signaling and improved TB protection. Cell Rep (2017) 18:2752–65.10.1016/j.celrep.2017.02.057
    1. Tameris MD, Hatherill M, Landry BS, Scriba TJ, Snowden MA, Lockhart S, et al. Safety and efficacy of MVA85A, a new tuberculosis vaccine, in infants previously vaccinated with BCG: a randomised, placebo-controlled phase 2b trial. Lancet (2013) 381:1021–8.10.1016/S0140-6736(13)60177-4
    1. Spertini F, Audran R, Chakour R, Karoui O, Steiner-Monard V, Thierry AC, et al. Safety of human immunisation with a live-attenuated Mycobacterium tuberculosis vaccine: a randomised, double-blind, controlled phase I trial. Lancet Respir Med (2015) 3:953–62.10.1016/S2213-2600(15)00435-X
    1. Andrews JR, Nemes E, Tameris M, Landry BS, Mahomed H, Mcclain JB, et al. Serial QuantiFERON testing and tuberculosis disease risk among young children: an observational cohort study. Lancet Respir Med (2017) 5:282–90.10.1016/S2213-2600(17)30060-7
    1. Tran V, Liu J, Behr MA. BCG vaccines. Microbiol Spectr (2014) 2:MGM2–28.10.1128/microbiolspec.MGM2-0028-2013
    1. Zhang L, Ru HW, Chen FZ, Jin CY, Sun RF, Fan XY, et al. Variable virulence and efficacy of BCG vaccine strains in mice and correlation with genome polymorphisms. Mol Ther (2016) 24:398–405.10.1038/mt.2015.216
    1. Liu J, Tran V, Leung AS, Alexander DC, Zhu B. BCG vaccines: their mechanisms of attenuation and impact on safety and protective efficacy. Hum Vaccin (2009) 5:70–8.10.4161/hv.5.2.7210
    1. Gonzalo-Asensio J, Malaga W, Pawlik A, Astarie-Dequeker C, Passemar C, Moreau F, et al. Evolutionary history of tuberculosis shaped by conserved mutations in the PhoPR virulence regulator. Proc Natl Acad Sci U S A (2014) 111:11491–6.10.1073/pnas.1406693111
    1. Broset E, Martin C, Gonzalo-Asensio J. Evolutionary landscape of the Mycobacterium tuberculosis complex from the viewpoint of PhoPR: implications for virulence regulation and application to vaccine development. MBio (2015) 6:e1289–1215.10.1128/mBio.01289-15
    1. Zhang W, Zhang Y, Zheng H, Pan Y, Liu H, Du P, et al. Genome sequencing and analysis of BCG vaccine strains. PLoS One (2013) 8:e71243.10.1371/journal.pone.0071243
    1. Nambiar JK, Pinto R, Aguilo JI, Takatsu K, Martin C, Britton WJ, et al. Protective immunity afforded by attenuated, PhoP-deficient Mycobacterium tuberculosis is associated with sustained generation of CD4+ T-cell memory. Eur J Immunol (2012) 42:385–92.10.1002/eji.201141903
    1. Yruela I, Contreras-Moreira B, Magalhaes C, Osorio NS, Gonzalo-Asensio J. Mycobacterium tuberculosis complex exhibits lineage-specific variations affecting protein ductility and epitope recognition. Genome Biol Evol (2016) 8:3751–64.10.1093/gbe/evw279
    1. Coscolla M, Copin R, Sutherland J, Gehre F, De Jong B, Owolabi O, et al. M. tuberculosis T cell epitope analysis reveals paucity of antigenic variation and identifies rare variable TB antigens. Cell Host Microbe (2015) 18:538–48.10.1016/j.chom.2015.10.008
    1. Andrews JR, Noubary F, Walensky RP, Cerda R, Losina E, Horsburgh CR. Risk of progression to active tuberculosis following reinfection with Mycobacterium tuberculosis. Clin Infect Dis (2012) 54:784–91.10.1093/cid/cir951

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