Mycobacterial codon optimization of the gene encoding the Sm14 antigen of Schistosoma mansoni in recombinant Mycobacterium bovis Bacille Calmette-Guérin enhances protein expression but not protection against cercarial challenge in mice.
A mycobacterial codon-optimized gene encoding the Sm14 antigen of Schistosoma mansoni was generated using oligonucleotide assembly. This synthetic gene enhanced approximately fourfold the protein expression level in recombinant Mycobacterium bovis Bacille Calmette-Guérin (rBCG) when compared to that obtained using the native gene in the same expression vector. Immunization of mice with rBCG expressing Sm14 via the synthetic gene induced specific cellular Th1-predominant immune responses, as determined by interferon-gamma production of Sm14-stimulated splenocytes, which were comparable to those recorded in animals immunized with an rBCG strain expressing the native gene. Administration of a single dose of the rBCG-Sm14 construct carrying the synthetic gene conferred protection against cercarial challenge in outbred Swiss mice, at a level equivalent to those provided by either a single dose of rBCG expressing the native gene or three doses of Escherichia coli-derived recombinant Sm14. Our data demonstrated that despite improving the level of antigen expression, the codon optimization strategy did not result in enhanced immunity or protection against cercarial S. mansoni challenge.
Varaldo
Paula B
PB
Centro de Biotecnologia, Instituto Butantan, Sao Paulo, Brazil.
Miyaji
Eliane N
EN
Vilar
Monica M
MM
Campos
Adriano S
AS
Dias
Waldely O
WO
Armôa
Geraldo R G
GR
Tendler
Miriam
M
Leite
Luciana C C
LC
McIntosh
Douglas
D
D000818:Animals; D001500:BCG Vaccine / Q000008:administration & dosage / Q000276:immunology*; D003062:Codon / Q000235:genetics; D050612:Fatty Acid Transport Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology* / Q000627:therapeutic use; D015870:Gene Expression / Q000187:drug effects*; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology* / Q000627:therapeutic use; D051379:Mice; D008807:Mice, Inbred BALB C; D009163:Mycobacterium bovis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D012550:Schistosoma mansoni / Q000737:chemistry* / Q000235:genetics; D012555:Schistosomiasis mansoni / Q000517:prevention & control*; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology; D014614:Vaccines, Synthetic
D001500:BCG Vaccine; D003062:Codon; D050612:Fatty Acid Transport Proteins; D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D014614:Vaccines, Synthetic; C068709:SM14 protein, Schistosoma mansoni