Article thumbnail

Molecular Characterization of Heterologous HIV-1gp120 Gene Expression Disruption in Mycobacterium bovis BCG Host Strain: A Critical Issue for Engineering Mycobacterial Based-Vaccine Vectors

By Joan Joseph, Raquel Fernández-Lloris, Elías Pezzat, Narcís Saubi, Pere-Joan Cardona, Beatriz Mothe and Josep Maria Gatell

Abstract

Mycobacterium bovis Bacillus Calmette-Guérin (BCG) as a live vector of recombinant bacterial vaccine is a promising system to be used. In this study, we evaluate the disrupted expression of heterologous HIV-1gp120 gene in BCG Pasteur host strain using replicative vectors pMV261 and pJH222. pJH222 carries a lysine complementing gene in BCG lysine auxotrophs. The HIV-1 gp120 gene expression was regulated by BCG hsp60 promoter (in plasmid pMV261) and Mycobacteria spp. α-antigen promoter (in plasmid pJH222). Among 14 rBCG:HIV-1gp120 (pMV261) colonies screened, 12 showed a partial deletion and two showed a complete deletion. However, deletion was not observed in all 10 rBCG:HIV-1gp120 (pJH222) colonies screened. In this study, we demonstrated that E. coli/Mycobacterial expression vectors bearing a weak promoter and lysine complementing gene in a recombinant lysine auxotroph of BCG could prevent genetic rearrangements and disruption of HIV 1gp120 gene expression, a key issue for engineering Mycobacterial based vaccine vectors

Topics: Research Article
Publisher: Hindawi Publishing Corporation
OAI identifier: oai:pubmedcentral.nih.gov:2896670
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles

Citations

  1. (2009). A novel insertion sequence, IS1642, of Mycobacterium avium, which forms long direct repeats of variable length: research letter,”FEMSMicrobiologyLetters,vol.291,no.2,pp.216–221,
  2. (2004). A Population, Health and Nutrition Information Project under USAID contract no.
  3. (1995). C i r i l l o ,C .K .S t o v e r ,B .R .B l o o m ,W .R .J a c o b sJ r . ,a n dR
  4. (2002). Combined intrarectal/intradermal inoculation of recombinant Mycobacterium bovis bacillus Calmette-Gu´ erin (BCG) induces enhanced immune responses against the inserted
  5. (1999). Comparison of the construction of unmarked deletion mutations in Mycobacterium smegmatis, Mycobacterium bovis bacillus Calmette-Gu´ erin, and Mycobacterium tuberculosis H37Rv by allelic exchange,”
  6. (2004). Cytotoxic T lymphocyte-based control of simian immunodeficiency virus replication in a preclinical AIDS vaccine trial,”
  7. Eds., Current Protocols in Molecular Biology,
  8. (1996). Efficient homologous recombination in fast-growing and slow-growing mycobacteria,”
  9. (2008). eller ,E.C.B¨ ottger,andP.Sander,“Tuberculosisvaccine strain Mycobacterium bovis BCG Russia is a natural recA mutant,”
  10. (1990). Establishment of a foreign antigen secretion system
  11. (2007). Evaluation of recombinant BCG expressing rotavirus VP6 as an anti-rotavirus vaccine,”
  12. (2002). Expression of foreign genes in Mycobacterium bovis BCG strains using different promoters reveals instability of the hsp60 promoter for expression of foreign genes
  13. (1991). Expression of heterologous genes in Mycobacterium bovis BCG: induction of a cellular response against
  14. (1991). F u e r s t ,C .K .S t o v e r ,a n dV .F .d el aC r u z ,“ D e v e l o p m e n t of BCG as a live recombinant vector system: potential use as an HIV vaccine,”
  15. (2005). Factors influencing the immuneresponsetoforeignantigenexpressedinrecombinant
  16. (2008). Gag-specific cytotoxic T-lymphocyte-based control of primary simian immunodeficiency virus replication in a vaccine trial,”
  17. (1998). Genetic rearrangements leading to disruption of heterologous gene expression in mycobacteria: an observation with Escherichia coli β-galactosidase in Mycobacterium smegmatis and its implication in vaccine development,”
  18. (2008). H u b e r ,M . - T .R u f ,G .P l u s c h k e ,a n dM .K¨ aser, “Independent loss of immunogenic proteins
  19. (1975). H.J.ReifandH.Saedler,“IS1isinvolvedindeletionformation in the gal region of E.
  20. (1991). Humoral and cell-mediated immune responses to live recombinant
  21. (2001). Instability and site-specific excision of integrationproficient mycobacteriophage L5 plasmids: development of stably maintained integrative vectors,”
  22. (2008). IS1096-mediated DNA rearrangements play a key role in genome evolution of Mycobacterium smegmatis,”
  23. (2008). IS6110, a Mycobacterium tuberculosis complex-specific insertion sequence, is also present in the genome of Mycobacterium smegmatis, suggestive of lateral gene transfer among mycobacterialspecies,”JournalofBacteriology,vol.190,no.9,pp.3408– 3410,
  24. (2001). P.Sander,K.G.Papavinasasundaram,T.Dicketal.,“Mycobacterium bovis BCG recA deletion mutant shows increased susceptibility to DNA-damaging agents but wild-type survival in a mouse infection model,”
  25. (2006). Progress towards an HIV vaccine based on recombinant Bacillus Calmette-Gu´ erin: failures and challenges,” Expert Review of Vaccines,
  26. (1996). Rearrangements of the bacterial chromosome formation and applications,”
  27. (1997). Recombinant Mycobacterium bovis BCG producing the N-terminal half of SIVmac251 Env antigen induces neutralizing antibodies and cytotoxic T lymphocyte responses in mice and guinea pigs,”
  28. (1988). Recombination between repeated genes in microorganisms,”
  29. (1999). S c h m i t z ,M .J .K u r o d a ,S .S a n t r ae ta l . ,“ C o n t r o lo f viremia in simian immunodeficiency virus infection by CD8+ lymphocytes,”
  30. (1991). S t o v e r ,V .F .d el aC r u z ,T .R .F u e r s te ta l . ,“ N e wu s eo f BCGforrecombinantvaccines,”Nature,vol.351,no.6326,pp. 456–460,
  31. (1991). Site-specific integration of mycobacteriophage L5: integration-proficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis, and bacille
  32. (1994). Structural instability of recombinant plasmids in mycobacteria,”
  33. (1994). The BCG experience: implications for future vaccines against tuberculosis,”
  34. (1988). The stability and immunogenicity of a dispersed-grown freeze-dried Pasteur BCG vaccine,”
  35. (1977). Transposable genetic elements as agents of gene instability and chromosomal rearrangements,”
  36. (1999). Transposition-induced structural instability of Escherichia coli-mycobacteria shuttle vectors,”
  37. (2007). Vaccine platform for prevention of tuberculosis and mother-to-child transmission of human immunodeficiency virus type 1 through breastfeeding,”
  38. (1997). Vigorous HIV-1-specific CD4+ T cell responses associated with control of viremia,”