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• W2912486588 abstract "The tuberculosis (TB) vaccine MTBVAC is the only live-attenuated Mycobacterium tuberculosis (Mtb)-based vaccine in clinical development, and it confers superior protection in different animal models compared to the current vaccine, BCG (Mycobacterium bovis bacillus Calmette-Guérin). With the aim of using MTBVAC as a vector for a dual TB-HIV vaccine, we constructed the recombinant MTBVAC.HIVA2auxo strain. First, we generated a lysine auxotroph of MTBVAC (MTBVACΔlys) by deleting the lysA gene. Then the auxotrophic MTBVACΔlys was transformed with the E. coli-mycobacterial vector p2auxo.HIVA, harboring the lysA-complementing gene and the HIV-1 clade A immunogen HIVA. This TB-HIV vaccine conferred similar efficacy to the parental strain MTBVAC against Mtb challenge in mice. MTBVAC.HIVA2auxo was safer than BCG and MTBVAC in severe combined immunodeficiency (SCID) mice, and it was shown to be maintained up to 42 bacterial generations in vitro and up to 100 days after inoculation in vivo. The MTBVAC.HIVA2auxo vaccine, boosted with modified vaccinia virus Ankara (MVA).HIVA, induced HIV-1 and Mtb-specific interferon-γ-producing T cell responses and polyfunctional HIV-1-specific CD8+ T cells producing interferon-γ (IFN-γ), tumor necrosis factor alpha (TNF-α), and CD107a in BALB/c mice. Here we describe new tools to develop combined vaccines against TB and HIV with the potential of expansion for other infectious diseases. The tuberculosis (TB) vaccine MTBVAC is the only live-attenuated Mycobacterium tuberculosis (Mtb)-based vaccine in clinical development, and it confers superior protection in different animal models compared to the current vaccine, BCG (Mycobacterium bovis bacillus Calmette-Guérin). With the aim of using MTBVAC as a vector for a dual TB-HIV vaccine, we constructed the recombinant MTBVAC.HIVA2auxo strain. First, we generated a lysine auxotroph of MTBVAC (MTBVACΔlys) by deleting the lysA gene. Then the auxotrophic MTBVACΔlys was transformed with the E. coli-mycobacterial vector p2auxo.HIVA, harboring the lysA-complementing gene and the HIV-1 clade A immunogen HIVA. This TB-HIV vaccine conferred similar efficacy to the parental strain MTBVAC against Mtb challenge in mice. MTBVAC.HIVA2auxo was safer than BCG and MTBVAC in severe combined immunodeficiency (SCID) mice, and it was shown to be maintained up to 42 bacterial generations in vitro and up to 100 days after inoculation in vivo. The MTBVAC.HIVA2auxo vaccine, boosted with modified vaccinia virus Ankara (MVA).HIVA, induced HIV-1 and Mtb-specific interferon-γ-producing T cell responses and polyfunctional HIV-1-specific CD8+ T cells producing interferon-γ (IFN-γ), tumor necrosis factor alpha (TNF-α), and CD107a in BALB/c mice. Here we describe new tools to develop combined vaccines against TB and HIV with the potential of expansion for other infectious diseases. Today, tuberculosis (TB) has reached alarming proportions. An estimated 10 million people have developed TB in 2017 and 9% were people living with HIV (72% in Africa). There were an estimated 1.3 million TB deaths among HIV-negative people and an additional 300,000 deaths among HIV-positive people, as reported by the World Health Organization (WHO)1WHOGlobal Tuberculosis Report 2018.https://www.who.int/tb/publications/global_report/en/Date: 2018Google Scholar in 2018. TB is poverty related with a major burden in the poor and developing parts of the world, and it is aggravated by the HIV-AIDS pandemic, which greatly increases the risk of the infection evolving into active TB disease. HIV-AIDS is a major global public health issue. Between 2010 and 2016, new HIV infections fell by 11% in adults and 47% in children, and AIDS-related deaths fell by 48% since the peak in 2005. This achievement was the result of great efforts by national HIV programs supported by civil society and a range of development partners.2WHOHIV/AIDS fact sheet.https://www.who.int/en/news-room/fact-sheets/detail/hiv-aidsDate: 2018Google Scholar However, sub-Saharan Africa accounted for 64% of new HIV infections in 2016, and, even though it is encouraging that 1.6 million people are currently receiving treatment in resource-poor settings, ensuring universal access to antiretroviral therapy still represents an enormous challenge.3UNAIDSFact sheet – World AIDS Day 2018.http://www.unaids.org/sites/default/files/media_asset/UNAIDS_FactSheet_en.pdfDate: 2018Google Scholar Thus, the development of effective, safe, and affordable vaccines against both diseases could have a tremendous impact on public health. The risk of active TB is estimated to be between 16 and 27 times greater in people living with HIV than among those without HIV infection.4WHOTuberculosis and HIV.https://www.who.int/hiv/topics/tb/about_tb/en/Date: 2015Google Scholar Mycobacterium bovis bacillus Calmette-Guérin (BCG) has been the only licensed vaccine against TB for more than 90 years,5Rodrigues L.C. Diwan V.K. Wheeler J.G. Protective effect of BCG against tuberculous meningitis and miliary tuberculosis: a meta-analysis.Int. J. Epidemiol. 1993; 22: 1154-1158Crossref PubMed Scopus (526) Google Scholar but the BCG-induced protective effects against pulmonary disease over all ages are variable.6Colditz G.A. Brewer T.F. Berkey C.S. Wilson M.E. Burdick E. Fineberg H.V. Mosteller F. Efficacy of BCG vaccine in the prevention of tuberculosis. Meta-analysis of the published literature.JAMA. 1994; 271: 698-702Crossref PubMed Scopus (1708) Google Scholar, 7Matsuo K. Yasutomi Y. Mycobacterium bovis Bacille Calmette-Guérin as a Vaccine Vector for Global Infectious Disease Control.Tuberc. Res. Treat. 2011; 2011: 574591PubMed Google Scholar Nevertheless, BCG8Fine P.E. Variation in protection by BCG: implications of and for heterologous immunity.Lancet. 1995; 346: 1339-1345Abstract PubMed Scopus (1094) Google Scholar vaccination has several beneficial effects: (1) BCG vaccination reduces rates of Mycobacterium tuberculosis (Mtb) infection, aiding in the decrease of the pool of latent infections from which future cases of active disease may arise;9Roy A. Eisenhut M. Harris R.J. Rodrigues L.C. Sridhar S. Habermann S. Snell L. Mangtani P. Adetifa I. Lalvani A. Abubakar I. Effect of BCG vaccination against Mycobacterium tuberculosis infection in children: systematic review and meta-analysis.BMJ. 2014; 349: g4643Crossref PubMed Scopus (352) Google Scholar (2) BCG provides strong protection against disseminated forms of the disease in infants and young children;10Mangtani P. Abubakar I. Ariti C. Beynon R. Pimpin L. Fine P.E.M. Rodrigues L.C. Smith P.G. Lipman M. Whiting P.F. Sterne J.A. Protection by BCG vaccine against tuberculosis: a systematic review of randomized controlled trials.Clin. Infect. Dis. 2014; 58: 470-480Crossref PubMed Scopus (564) Google Scholar, 11Graham S.M. Sismanidis C. Menzies H.J. Marais B.J. Detjen A.K. Black R.E. Importance of tuberculosis control to address child survival.Lancet. 2014; 383: 1605-1607Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar (3) BCG revaccination of adolescents may provide additional benefits for the prevention of TB;12Nemes E. Geldenhuys H. Rozot V. Rutkowski K.T. Ratangee F. Bilek N. Mabwe S. Makhethe L. Erasmus M. Toefy A. et al.C-040-404 Study TeamPrevention of M. tuberculosis Infection with H4:IC31 Vaccine or BCG Revaccination.N. Engl. J. Med. 2018; 379: 138-149Crossref PubMed Scopus (366) Google Scholar and (4) BCG vaccination reduces all-cause mortality through beneficial non-specific (heterologous) effects on the immune system.13Freyne B. Marchant A. Curtis N. BCG-associated heterologous immunity, a historical perspective: experimental models and immunological mechanisms.Trans. R. Soc. Trop. Med. Hyg. 2015; 109: 46-51Crossref PubMed Scopus (30) Google Scholar, 14Arts R.J.W. Moorlag S.J.C.F.M. Novakovic B. Li Y. Wang S.-Y. Oosting M. Kumar V. Xavier R.J. Wijmenga C. Joosten L.A.B. et al.BCG Vaccination Protects against Experimental Viral Infection in Humans through the Induction of Cytokines Associated with Trained Immunity.Cell Host Microbe. 2018; 23: 89-100.e5Abstract Full Text Full Text PDF PubMed Scopus (612) Google Scholar These four effects strengthen the motivation for the inclusion of BCG in the global vaccination program.15Marais B.J. Seddon J.A. Detjen A.K. van der Werf M.J. Grzemska M. Hesseling A.C. Curtis N. Graham S.M. WHO Child TB SubgroupInterrupted BCG vaccination is a major threat to global child health.Lancet Respir. Med. 2016; 4: 251-253Abstract Full Text Full Text PDF PubMed Scopus (25) Google Scholar We previously constructed and characterized MTBVAC, the first and only live-attenuated Mtb-based vaccine candidate in clinical development against TB disease in the pipeline. MTBVAC contains two independent deletions in the phoP and fadD26 genes without antibiotic resistance markers, and it fulfills the Geneva consensus requirements for progressing into clinical trials.16Arbues A. Aguilo J.I. Gonzalo-Asensio J. Marinova D. Uranga S. Puentes E. Fernandez C. Parra A. Cardona P.J. Vilaplana C. et al.Construction, characterization and preclinical evaluation of MTBVAC, the first live-attenuated M. tuberculosis-based vaccine to enter clinical trials.Vaccine. 2013; 31: 4867-4873Crossref PubMed Scopus (167) Google Scholar The vaccine candidate MTBVAC was safe, and it conferred superior protection in different animal models compared to the licensed BCG reference strain in use today. To date, phase I trials in adults and neonates (ClinicalTrials.gov: NCT02013245 and NCT02729571) have been successfully completed, and phase II trials for dose definition, at birth and in adults with and without latent TB, are in progress (ClinicalTrials.gov: NCT02933281 and NCT03536117). Clinical results showed that MTBVAC was immunogenic, in a dose-dependent manner, and it had a similar safety profile as that of BCG.16Arbues A. Aguilo J.I. Gonzalo-Asensio J. Marinova D. Uranga S. Puentes E. Fernandez C. Parra A. Cardona P.J. Vilaplana C. et al.Construction, characterization and preclinical evaluation of MTBVAC, the first live-attenuated M. tuberculosis-based vaccine to enter clinical trials.Vaccine. 2013; 31: 4867-4873Crossref PubMed Scopus (167) Google Scholar, 17Spertini F. Audran R. Chakour R. Karoui O. Steiner-Monard V. Thierry A.-C. Mayor C.E. Rettby N. Jaton K. Vallotton L. 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: 953-962Abstract Full Text Full Text PDF PubMed Scopus (116) Google Scholar It is well known that there is strong evidence in favor of a role for HIV-1-specific T cell responses in the control of HIV-1 replication.18Koup R.A. Safrit J.T. Cao Y. Andrews C.A. McLeod G. Borkowsky W. Farthing C. Ho D.D. Temporal association of cellular immune responses with the initial control of viremia in primary human immunodeficiency virus type 1 syndrome.J. Virol. 1994; 68: 4650-4655Crossref PubMed Google Scholar, 19Rowland-Jones S.L. Dong T. Fowke K.R. Kimani J. Krausa P. Newell H. Blanchard T. Ariyoshi K. Oyugi J. Ngugi E. et al.Cytotoxic T cell responses to multiple conserved HIV epitopes in HIV-resistant prostitutes in Nairobi.J. Clin. Invest. 1998; 102: 1758-1765Crossref PubMed Scopus (377) Google Scholar One promising approach for T cell induction is M. bovis BCG as a bacterial live recombinant vaccine vehicle. Specific humoral and cellular immune responses against HIV-1 have been detected after immunization of mice with recombinant BCG (rBCG) expressing HIV-1 antigens.20Stover C.K. de la Cruz V.F. Fuerst T.R. Burlein J.E. Benson L.A. Bennett L.T. Bansal G.P. Young J.F. Lee M.H. Hatfull G.F. et al.New use of BCG for recombinant vaccines.Nature. 1991; 351: 456-460Crossref PubMed Scopus (1205) Google Scholar, 21Aldovini A. Young R.A. Humoral and cell-mediated immune responses to live recombinant BCG-HIV vaccines.Nature. 1991; 351: 479-482Crossref PubMed Scopus (253) Google Scholar, 22Lagranderie M. Murray A. Gicquel B. Leclerc C. Gheorghiu M. Oral immunization with recombinant BCG induces cellular and humoral immune responses against the foreign antigen.Vaccine. 1993; 11: 1283-1290Crossref PubMed Scopus (63) Google Scholar, 23Kim B.-J. Kim B.-R. Kook Y.-H. Kim B.-J. Development of a Live Recombinant BCG Expressing Human Immunodeficiency Virus Type 1 (HIV-1) Gag Using a pMyong2 Vector System: Potential Use As a Novel HIV-1 Vaccine.Front. Immunol. 2018; 9: 643Crossref PubMed Scopus (13) Google Scholar, 24Venkataswamy M.M. Ng T.W. Kharkwal S.S. Carreño L.J. Johnson A.J. Kunnath-Velayudhan S. Liu Z. Bittman R. Jervis P.J. Cox L.R. et al.Improving Mycobacterium bovis bacillus Calmette-Guèrin as a vaccine delivery vector for viral antigens by incorporation of glycolipid activators of NKT cells.PLoS One. 2014; 9: e108383Crossref PubMed Scopus (21) Google Scholar We previously developed several rBCG HIV-1 vaccine candidates with the aim of inducing protective cell-mediated responses. Confirming the efficacy of an HIV-1 vaccine candidate in humans is as of yet not possible in animal studies alone. Achieving protection against HIV infection in humans following active vaccination, and subsequently identifying the correlates of protection, would allow the validation of protection in animal models. Our aim is to induce a strong CD8+ T cell response capable of aiding and complementing the protective efficacy of antibody-based vaccines, while providing viral control in the case of an infection occurring. Our starting platform was based on a heterologous rBCG prime and recombinant modified vaccinia virus Ankara (MVA) boost regimen delivering a common immunogen called HIVA (MVA.HIVA), which is derived from consensus Gag protein of HIV-1 clade A, prevalent in central and eastern Africa, and a string of human CD8+ T cell epitopes.25Hanke T. McMichael A.J. Design and construction of an experimental HIV-1 vaccine for a year-2000 clinical trial in Kenya.Nat. Med. 2000; 6: 951-955Crossref PubMed Scopus (185) Google Scholar Recently, we engineered a new BCG.HIVA2auxo vaccine strain harboring an antibiotic-free plasmid selection system and maintenance. The BCG.HIVA2auxo vaccine in combination with MVA.HIVA was safe, and it induced HIV-1 and Mtb-specific interferon-γ-producing T cell responses in adult BALB/c mice.26Saubi N. Gea-Mallorquí E. Ferrer P. Hurtado C. Sánchez-Úbeda S. Eto Y. Gatell J.M. Hanke T. Joseph J. Engineering new mycobacterial vaccine design for HIV-TB pediatric vaccine vectored by lysine auxotroph of BCG.Mol. Ther. Methods Clin. Dev. 2014; 1: 14017Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar In this study we have constructed a novel live-attenuated vaccine for HIV-1 and TB that is vectored by a lysine auxotroph of MTBVAC, MTBVAC.HIVA2auxo. This is an innovative approach to develop bivalent TB and HIV vaccines that could be administered at birth and with the potential to confer protection against both diseases. For MTBVACΔlys construction, the previously described recombineering-based technique was used.27Aguilo N. Gonzalo-Asensio J. Alvarez-Arguedas S. Marinova D. Gomez A.B. Uranga S. Spallek R. Singh M. Audran R. Spertini F. Martin C. Reactogenicity to major tuberculosis antigens absent in BCG is linked to improved protection against Mycobacterium tuberculosis.Nat. Commun. 2017; 8: 16085Crossref PubMed Scopus (78) Google Scholar Rv1293 (lysA) gene, which codes for the last enzyme involved in Lysine (Lys) synthesis,28Pavelka Jr., M.S. Jacobs Jr., W.R. Comparison of the construction of unmarked deletion mutations in Mycobacterium smegmatis, Mycobacterium bovis bacillus Calmette-Guérin, and Mycobacterium tuberculosis H37Rv by allelic exchange.J. Bacteriol. 1999; 181: 4780-4789Crossref PubMed Google Scholar was inactivated by homologous recombination with a PCR product containing a kanamycin (Km) resistance cassette disturbing the lysA gene (Figure 1A). To ensure proper selection of recombinants, the final MTBVAC transformed with the homologous PCR product lysA-Km was plated on 7H10 complete medium supplemented with Km and Lys. Correct recombination was confirmed by PCR using three different pairs of primers, which amplify the complete recombined region (Lys-fw/Lys-rv), the upstream (Lys-fw/km-OUT1-rv), or the downstream (km-OUT2-fw/Lys-rv) region (Figure 1B). After MTBVACΔlys construction, Lys auxotrophy was confirmed by plating on 7H10-ADC with and without Lys supplementation (Figure 1C) and also by colony-forming unit (CFU) enumeration after removing Lys from medium (data not shown). Results showed the absence of MTBVACΔlys growth in non-lysine-supplemented plates, whereas the MTBVAC strain grew at a similar level in both supplemented and non-supplemented plates. Accordingly, this auxotrophic MTBVACΔlys strain was used in the subsequent experiments to generate recombinant vaccines expressing the HIVA immunogen. The plasmid p2auxo.HIVA (Figure 2A)26Saubi N. Gea-Mallorquí E. Ferrer P. Hurtado C. Sánchez-Úbeda S. Eto Y. Gatell J.M. Hanke T. Joseph J. Engineering new mycobacterial vaccine design for HIV-TB pediatric vaccine vectored by lysine auxotroph of BCG.Mol. Ther. Methods Clin. Dev. 2014; 1: 14017Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar was transformed into the MTBVACΔlys host strain to generate the recombinant MTBVAC.HIVA2auxo. The selection of positive recombinant MTBVAC.HIVA2auxo colonies was performed by culturing the MTBVACΔlys transformants on Middlebrook agar 7H10 medium without lysine supplementation. The MTBVAC.HIVA2auxo strain harboring the lysine-complementing gene abolished the requirement for exogenous lysine, and colonies were observed in non-lysine-supplemented agar plates (Figure 2B). The expression of the full-size chimeric 19-kDa signal sequence-HIVA protein (total weight, 64 kDa) was confirmed by western blot analysis of the MTBVAC.HIVA2auxo cell lysates (Figure 2C). No HIVA protein expression was detected in recombinant MTBVAC strains harboring the p2auxo plasmid without heterologous insert (MTBVAC.∅2auxo, negative control). This proper molecular characterization led us to prepare a master seed stock and derivative working vaccine stock for downstream experiments. To assess the in vitro stability of the p2auxo.HIVA plasmid, subcultures of MTBVAC.HIVA2auxo on selective media (no lysine supplementation) were carried out every 7 days. The maintenance of the p2auxo.HIVA plasmid DNA was evaluated by PCR analysis of HIVA and GlyA DNA-coding sequences. Bands corresponding to the HIVA DNA-coding sequence (Figure 3A) and to the E. coli GlyA-coding sequence (Figure S1A) were observed in all 6 MTBVAC.HIVA2auxo subcultures (42 bacterial generations), indicating that there were no major genetic rearrangements in the HIVA and glyA genes of MTBVAC.HIVA2auxo vaccine strain over the subsequent subculturing passages. In vivo stability of p2auxo.HIVA plasmid in MTBVAC.HIVA2auxo was assessed in severe combined immunodeficiency (SCID) mice used in the safety trial. Homogenized spleens were plated on Lys-supplemented medium, and p2auxo.HIVA presence in the mycobacterial burden was analyzed by colony PCR using primers to detect the HIVA DNA-coding sequence (Figure 3B) and the glyA gene (Figure S1B). The analysis showed that 95.5% of the colonies retained the plasmid during in vivo infection. We previously demonstrated that heterologous BCG.HIVA prime boosted with MVA.HIVA elicited high-quality HIV-1-specific T cell responses.26Saubi N. Gea-Mallorquí E. Ferrer P. Hurtado C. Sánchez-Úbeda S. Eto Y. Gatell J.M. Hanke T. Joseph J. Engineering new mycobacterial vaccine design for HIV-TB pediatric vaccine vectored by lysine auxotroph of BCG.Mol. Ther. Methods Clin. Dev. 2014; 1: 14017Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar, 29Im E.-J. Saubi N. Virgili G. Sander C. Teoh D. Gatell J.M. McShane H. Joseph J. Hanke T. Vaccine platform for prevention of tuberculosis and mother-to-child transmission of human immunodeficiency virus type 1 through breastfeeding.J. Virol. 2007; 81: 9408-9418Crossref PubMed Scopus (42) Google Scholar, 30Mahant A. Saubi N. Eto Y. Guitart N. Gatell J.M. Hanke T. Joseph J. Preclinical development of BCG.HIVA2auxo.int, harboring an integrative expression vector, for a HIV-TB Pediatric vaccine. Enhancement of stability and specific HIV-1 T-cell immunity.Hum. Vaccin. Immunother. 2017; 13: 1798-1810Crossref PubMed Scopus (12) Google Scholar In this study, we evaluated the specific HIV-1 T cell responses in adult BALB/c mice after intradermal immunization with MTBVAC.HIVA2auxo or MTBVAC.Ø2auxo prime and intramuscular MVA.HIVA boost (Figure 4A). The intradermal route mimics the administration performed in human BCG vaccination, and it has been shown to elicit a higher HIV-1-specific CD8+ T cell response in adult BALB/c mice.31Saubi N. Im E.-J. Fernández-Lloris R. Gil O. Cardona P.-J. Gatell J.M. Hanke T. Joseph J. Newborn mice vaccination with BCG.HIVA222 + MVA.HIVA enhances HIV-1-specific immune responses: influence of age and immunization routes.Clin. Dev. Immunol. 2011; 2011: 516219Crossref PubMed Scopus (19) Google Scholar The immunogenicity readout was focused on the P18-I10 epitope, an immunodominant cytotoxic T-lymphocytes (CTL) epitope derived from HIV-1 Env and H-2Dd murine restricted, which was fused to HIVA immunogen to evaluate the immunogenicity in mice. On day 0, adult mice were either left unimmunized or primed with MTBVAC.HIVA2auxo or MTBVAC.Ø2auxo, and on week 6 the animals received an MVA.HIVA boost. Mice were sacrificed on week 8, and the functional quality of the elicited CD8+ T cells to produce interferon-γ (IFN-γ) and tumor necrosis factor alpha (TNF-α) and to degranulate (surface expression of CD107a) in response to P18-I10 peptide stimulation was measured by intracellular cytokine staining (ICS) (Figure 4B). We observed in adult mice that MTBVAC.HIVA2auxo prime and MVA.HIVA boost induced higher frequencies of P18-I10 epitope-specific CD8+ splenocytes producing IFN-γ, TNF-α, and CD107 than mice primed with MTBVAC.Ø2auxo, MVA.HIVA alone, or naive mice. We found that MTBVAC.HIVA2auxo prime and MVA.HIVA boost induced higher frequencies of trifunctional specific CD8+ T cells compared with the MTBVAC.Ø2auxo priming and MVA.HIVA boost and with MVA.HIVA alone (Figure 4C). The capacity of splenocytes from vaccinated mice to secrete IFN-γ was also assessed by the enzyme-linked immunosorbent spot (ELISPOT) assay. We observed the highest frequency of specific cells secreting IFN-γ when stimulated with P18-I10 in mice primed with MTBVAC.HIVA2auxo and boosted with MVA.HIVA, 1,280 spot-forming units (SFU)/106 splenocytes, compared to 1,043 SFU/106 splenocytes obtained when mice were primed with MTBVAC.Ø2auxo and 1,095 SFU/106 splenocytes when mice were only boosted with MVA.HIVA (Figure 4D). The capacity of splenocytes from vaccinated mice to secrete IFN-γ after overnight stimulation with the Mtb-purified protein derivative (PPD) was also assessed by ELISPOT. The median SFUs per 106 splenocytes were similar in mice primed with MTBVAC.HIVA2auxo and MTBVAC.Ø2auxo (102 and 86 SFU/106 splenocytes, respectively; Figure 4E). As shown in Figure 5, the body mass was monitored over time and recorded to depict any adverse events and body mass loss due to vaccination. To detect vaccine-derived adverse events, a 12-week period between MTBVAC.HIVA2auxo and MVA.HIVA boost was established for this trial. Importantly, no statistically significant difference (by ANOVA) was observed between the vaccinated mouse groups and the control mouse group at the final time point. Furthermore, between weeks 1 and 14, the body mass monitored in all vaccinated mouse groups was found to lie between the mean ± 2 SD body mass curves in control mice. It is also important to mention that no mice died during the trial, and no local adverse events or associated systemic reactions were observed. We evaluated the efficacy of the bivalent vaccine strain MTBVAC.HIVA2auxo with respect to the parental MTBVAC vaccine. Groups of 6 C57BL/6 mice were left unimmunized or vaccinated with MTBVAC, MTBVAC.HIVA2auxo, or MTBVACΔlys by subcutaneous injection, a route previously used for efficacy studies in mouse models.32Zhang L. Ru H.-W. Chen F.-Z. Jin C.-Y. Sun R.-F. Fan X.-Y. Guo M. Mai J.T. Xu W.X. Lin Q.X. Liu J. Variable Virulence and Efficacy of BCG Vaccine Strains in Mice and Correlation With Genome Polymorphisms.Mol. Ther. 2016; 24: 398-405Abstract Full Text Full Text PDF PubMed Scopus (74) Google Scholar At 7 weeks post-vaccination, the mice were challenged with the pathogenic H37Rv strain by the intranasal route. Bacterial load in lungs and spleens was examined 4 weeks post-challenge by plating homogenized organs on complete 7H10 medium (Figure 6). In all vaccinated groups, the bacterial reduction was significant with respect to the unvaccinated group, both in lungs and spleens. The auxotrophic strain MTBVACΔlys, which was expected not to be able to survive without lysine, also displayed significant protection against Mtb H37Rv when compared to the naive group. No differences were found between the different MTBVAC strains tested, which validates the protective behavior of MTBVAC.HIVA2auxo vaccine against Mtb despite the genetic manipulations introduced. As well as affecting vaccine efficacy, genetic manipulation may also affect attenuation of live vaccines.32Zhang L. Ru H.-W. Chen F.-Z. Jin C.-Y. Sun R.-F. Fan X.-Y. Guo M. Mai J.T. Xu W.X. Lin Q.X. Liu J. Variable Virulence and Efficacy of BCG Vaccine Strains in Mice and Correlation With Genome Polymorphisms.Mol. Ther. 2016; 24: 398-405Abstract Full Text Full Text PDF PubMed Scopus (74) Google Scholar With the aim of corroborating the attenuation status of MTBVAC.HIVA2auxo and MTBVACΔlys strains, SCID mice were inoculated with 106 CFU by the intraperitoneal route, and the survival of animals was monitored (Figure 7). SCID mice are the reference model for safety assessments of live vaccines in preclinical TB studies, as recommended by regulatory bodies.33Walker K.B. Brennan M.J. Ho M.M. Eskola J. Thiry G. Sadoff J. Dobbelaer R. Grode L. Liu M.A. Fruth U. Lambert P.H. The second Geneva Consensus: Recommendations for novel live TB vaccines.Vaccine. 2010; 28: 2259-2270Crossref PubMed Scopus (85) Google Scholar Intraperitoneal, as well as intravenous, administration is a systemic inoculation route that allows rapid dissemination of the bacteria and, thereby, virulence assessments.32Zhang L. Ru H.-W. Chen F.-Z. Jin C.-Y. Sun R.-F. Fan X.-Y. Guo M. Mai J.T. Xu W.X. Lin Q.X. Liu J. Variable Virulence and Efficacy of BCG Vaccine Strains in Mice and Correlation With Genome Polymorphisms.Mol. Ther. 2016; 24: 398-405Abstract Full Text Full Text PDF PubMed Scopus (74) Google Scholar The auxotrophic MTBVACΔlys strain showed a hyper-attenuated profile; all mice inoculated with this strain survived until the endpoint of the experiment at week 31. Bacterial burden per spleen of these SCID mice vaccinated with MTBVACΔlys at week 31 was approximately 5 × 103 CFU (Figure S2), which demonstrated that this strain survived in vivo. When we analyzed survival time of MTBVAC.HIVA2auxo-vaccinated mice, data revealed a marked attenuation profile (they survived approximately 160 days) when compared to the BCG-vaccinated mice (deceased by day 90) and the MTBVAC-vaccinated mice (deceased by day 120). Despite the progress made in the development of a safe, effective, and affordable vaccine against HIV-1 and TB shortly after birth, the prevention of mother-to-child HIV-1 transmission via breast milk and childhood TB still remain great challenges. The use of mycobacteria as a vaccine vector is an attractive option; on top of the previously mentioned advantages (cheap mass production, good safety profile, suitable for neonates, etc.), it induces a potent Th1 type immune response (the central defense mechanism against intracellular pathogens) in humans and mice.34Ota M.O.C. Vekemans J. Schlegel-Haueter S.E. Fielding K. Sanneh M. Kidd M. Newport M.J. Aaby P. Whittle H. Lambert P.H. et al.Influence of Mycobacterium bovis bacillus Calmette-Guérin on antibody and cytokine responses to human neonatal vaccination.J. Immunol. 2002; 168: 919-925Crossref PubMed Scopus (249) Google Scholar, 35Marchant A. Goetghebuer T. Ota M.O. Wolfe I. Ceesay S.J. De Groote D. Corrah T. Bennett S. Wheeler J. Huygen K. et al.Newborns develop a Th1-type immune response to Mycobacterium bovis bacillus Calmette-Guérin vaccination.J. Immunol. 1999; 163: 2249-2255PubMed Google Scholar Three experimental systems must be orchestrated to develop a recombinant Mycobacterium-based HIV vaccine: (1) a live vaccine vehicle based on mycobacteria, (2) an E. coli-mycobacterial expression vector without antibiotic resistance markers, and (3) an HIV immunogen design. In this study, we have engineered a novel live-attenuated vaccine for HIV-1 and Mtb infection that is vectored by a lysine auxotroph of MTBVAC,16Arbues A. Aguilo J.I. Gonzalo-Asensio J. Marinova D. Uranga S. Puentes E. Fernandez C. Parra A. Cardona P.J. Vilaplana C. et al.Construction, characterization and preclinical evaluation of MTBVAC, the first live-attenuated M. tuberculosis-based vaccine to enter clinical trials.Vaccine. 2013; 31: 4867-4873Crossref PubMed Scopus (167) Google Scholar which expresses the HIV-1 clade A-derived immunogen HIVA.25Hanke T. McMichael A.J. Design and construction of an experimental HIV-1 vaccine for a year-2000 clinical trial in Kenya.Nat. Med. 2000; 6: 951-955Crossref PubMed Scopus (185) Google Scholar The use of mycobacterial vectors20Stover C.K. de la Cruz V.F. Fuerst T.R. Burlein J.E. Benson L.A. Bennett L.T. Bansal G.P. Young J.F. Lee M.H. Hatfull G.F. et al.New use of BCG for recombinant vaccines.Nature. 1991; 351: 456-460Crossref PubMed Scopus (1205) Google Scholar," @default.
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• W2912486588 cites W2171499092 @default.
• W2912486588 cites W2171843497 @default.
• W2912486588 cites W2176522783 @default.
• W2912486588 cites W2239561092 @default.
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