Mycobacterial Codon Optimization Enhances Antigen Expression and Virus-Specific Immune Responses in Recombinant<i>Mycobacterium bovis</i>Bacille Calmette-Guérin Expressing Human Immunodeficiency Virus Type 1 Gag

Kanekiyo, Masaru; Matsuo, Kazuhiro; Hamatake, Makiko; Hamano, Takaichi; Ohsu, Takeaki; Matsumoto, Sohkichi; Yamada, Takeshi; Yamazaki, Shudo; Hasegawa, Atsuhiko; Yamamoto, Naoki; Honda, Mitsuo

doi:10.1128/jvi.79.14.8716-8723.2005

Cited by 32 publications

(27 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Indeed, the result that the codon-usage adapted flp m gene confers a much higher recombination efficiency both in M. smegmatis and M. bovis BCG than the flp e gene of S. cerevisiae, strongly indicates that mRNA transcribed from the flp e gene of S. cerevisiae was not efficiently translated in both organisms. This is consistent with previous observations that codon usage adaptation improved the expression of other heterologous genes with low G + C content in mycobacteria, such as Sm14 antigen of Schistosoma mansoni (Varaldo et al, 2006) and human immunodeficiency virus type 1 Gag (Kanekiyo et al, 2005). Taken together, these studies indicate that codon usage adaptation may be a generally useful approach to increase expression of heterologous genes in mycobacteria.…”

Section: Role Of Codon Usage For the Expression Of Heterologous Genessupporting

confidence: 90%

Functional expression of the Flp recombinase in Mycobacterium bovis BCG

Song

Niederweis

2007

Gene

View full text Add to dashboard Cite

Mycobacteria contain a large number of redundant genes whose functions are difficult to analyze in mutants because there are only two efficient antibiotic resistance genes available for allelic exchange experiments. Sequence-specific recombinbases such as the Flp recombinase can be used to excise resistance markers. Expression of the flp(e) gene from Saccharomyces cerevisiae is functional for this purpose in fast-growing Mycobacterium smegmatis but not in slow-growing mycobacteria such as M. bovis BCG or M. tuberculosis. We synthesized the flp(m) gene by adapting the codon usage to that preferred by M. tuberculosis. This increased the G+C content from 38% to 61%. Using the synthetic flp(m) gene, the frequency of removal of FRT-hyg-FRT cassette from the chromosome by the Flp recombinase was increased by more than 100-fold in M. smegmatis. In addition, 40% of all clones of M. bovis BCG had lost the hyg resistance cassette after transient expression of the flp(m) gene. Sequencing of the chromosomal DNA showed that excision of the FRT-hyg-FRT cassette by Flp was specific. These results show that the flp(m) encoded Flp recombinase is not only an improved genetic tool for M. smegmatis, but can also be used in slow growing mycobacteria such as M. tuberculosis for constructing unmarked mutations. Other more sophisticated applications in mycobacterial genetics would also profit from the improved Flp/FRT system.

show abstract

Section: Role Of Codon Usage For the Expression Of Heterologous Genessupporting

confidence: 90%

Functional expression of the Flp recombinase in Mycobacterium bovis BCG

Song

Niederweis

2007

Gene

View full text Add to dashboard Cite

show abstract

“…However, this did not translate into an increased immune response (IFNg secretion by splenocytes stimulated in vitro with rSm14) or protective efficacy. This contrasts with an earlier report in which an rBCG strain expressing codon-optimized HIV Gag was more immunogenic than an rBCG expressing wildtype Gag (~40-fold increase in expression for the codon-optimized Gag) [99].…”

Section: Parasitic Diseasescontrasting

confidence: 69%

New Generation BCG Vaccines

Tullius

Horwitz

2010

Replicating Vaccines

View full text Add to dashboard Cite

Bacille de Calmette et Guérin (BCG) was attenuated from a virulent strain of Mycobacterium bovis a century ago and has since been administered as an anti-tuberculosis (TB) vaccine to more than four billion people, making it the most widely used vaccine of all time. Although BCG provides significant protection against disease and death due to childhood and disseminated forms of TB, the efficacy of BCG against adult, pulmonary disease is inconsistent. Thus, despite near universal vaccination with BCG in TB endemic areas, TB remains a heavy burden worldwide, especially in developing nations. In recent years, BCG has been utilized in two major vaccine development strategies. First, BCG has been used as a vector to express foreign antigens in studies aimed at developing new vaccines against a variety of viral, parasitic, and bacterial pathogens, and against cancer and allergic diseases. More recently, in a new vaccine paradigm, BCG has been used as a homologous vector to overexpress native mycobacterial antigens in studies aimed at developing improved vaccines against TB. As a vaccine vector, BCG has several major advantages including a very well-established safety profile, high immunogenicity (excellent CD4þ and CD8þ T-cell responses and strong TH1-Type immune responses), and low manufacturing cost. As a vector for recombinant TB vaccines, BCG has the additional advantages of providing a broad array of relevant mycobacterial antigens in addition to the recombinant antigens, moderate efficacy to begin with, high acceptability as a replacement vaccine for BCG in TB endemic countries, and the capacity to express M. tuberculosis proteins in native form and release them in a way that results in their being processed similarly to M. tuberculosis proteins. In addition to the overexpression of native proteins to improve their immunogenicity and protective efficacy against TB, recombinant BCG vaccines have been developed that express immunomodulatory cytokines or have been

show abstract

“…As most mammalian genes favour a different codon usage it is not surprising that codon optimisation can increase foreign gene expression in rBCG. This is reflected by an increase in immunogenicity of rBCG expressing codon optimised p24 compared to native p24 [122]. However, there are also some examples of genes with low G+C content that are expressed at high levels in BCG, such as ospA of Borrelia burgdorferi (30% G+C) [11] and Streptococcus pneumoniae pspA (38% G+C) [123].…”

Section: Factors To Consider When Designing a Rbcg Vaccine Vectormentioning

confidence: 99%

Recombinant Mycobacterium bovis BCG as an HIV Vaccine Vector

Chapman

Chege²,

Shephard³

et al. 2010

CHR

View full text Add to dashboard Cite

HIV-1 has resulted in a devastating AIDS pandemic. An effective HIV/AIDS vaccine that can be used to either, prevent HIV infection, control infection or prevent progression of the disease to AIDS is needed. In this review we discuss the use of Mycobacterium bovis BCG, the tuberculosis vaccine, as a vaccine vector for an HIV vaccine. Numerous features make BCG an attractive vehicle to deliver HIV antigens. It has a good safety profile, elicits long-lasting cellular immune responses and in addition manufacturing costs are affordable, a necessary consideration for developing countries. In this review we discuss the numerous factors that influence generation of a genetically stable recombinant BCG vaccine for HIV.

show abstract

Mycobacterial Codon Optimization Enhances Antigen Expression and Virus-Specific Immune Responses in RecombinantMycobacterium bovisBacille Calmette-Guérin Expressing Human Immunodeficiency Virus Type 1 Gag

Cited by 32 publications

References 42 publications

Functional expression of the Flp recombinase in Mycobacterium bovis BCG

Functional expression of the Flp recombinase in Mycobacterium bovis BCG

New Generation BCG Vaccines

Recombinant Mycobacterium bovis BCG as an HIV Vaccine Vector

Contact Info

Product

Resources

About