Evaluation of mammalian codon usage of fimH in DNA vaccine design

Bagherpour, Ghasem; Fooladi, Abbas Ali Imani; Mehrabadi, Jalil Fallah; Nourani, Mohammad Reza; Einollahi, Behzad

doi:10.1556/amicr.58.2011.4.2

Cited by 5 publications

(6 citation statements)

References 40 publications

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“…In addition to the use of optimized codons, several studies [32–34] have also reported that the expression and ELISA titer of synthesized genes markedly increased upon the removal of rare codons and unstable sequences. Bagherpour et al synthesized fimH of Uropathogenic Escherichia coli ( UPEC ) using mammalian codons [35]. Compared with the wild-type gene, the fimH gene of the mammalian codon has been shown to be compatible with eukaryotic expression systems.…”

Section: Discussionmentioning

confidence: 99%

Codon Preference Optimization Increases Prokaryotic Cystatin C Expression

Wang

Cui²,

Zhen³

et al. 2012

Journal of Biomedicine and Biotechnology

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Gene expression is closely related to optimal vector-host system pairing in many prokaryotes. Redesign of the human cystatin C (cysC) gene using the preferred codons of the prokaryotic system may significantly increase cysC expression in Escherichia coli (E. coli). Specifically, cysC expression may be increased by removing unstable sequences and optimizing GC content. According to E. coli expression system codon preferences, the gene sequence was optimized while the amino acid sequence was maintained. The codon-optimized cysC (co-cysC) and wild-type cysC (wt-cysC) were expressed by cloning the genes into a pET-30a plasmid, thus transforming the recombinant plasmid into E. coli BL21. Before and after the optimization process, the prokaryotic expression vector and host bacteria were examined for protein expression and biological activation of CysC. The recombinant proteins in the lysate of the transformed bacteria were purified using Ni2+-NTA resin. Recombinant protein expression increased from 10% to 46% based on total protein expression after codon optimization. Recombinant CysC purity was above 95%. The significant increase in cysC expression in E. coli expression produced by codon optimization techniques may be applicable to commercial production systems.

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Section: Discussionmentioning

confidence: 99%

Codon Preference Optimization Increases Prokaryotic Cystatin C Expression

Wang

Cui²,

Zhen³

et al. 2012

Journal of Biomedicine and Biotechnology

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“…Plasmids pVAX/ fim H(mam) and pVAX/ fim H(wt) were constructed in our laboratory as described previously [14]. Briefly, fim H(mam) comprised of mammalian (mam) codon usage sequences of Fim H protein and fim H(wt) composed of wild type (wt) codon usage sequences of FimH protein.…”

Section: Methodsmentioning

confidence: 99%

“…Electoporation delivery system was used to enhance host immune responses. In an independent study [14], we confirmed the expression of current DNA vaccine in vitro ; in the present study, we intended to examine the induction of cellular immunity against recurrent UTI in response to DNA vaccine in BALB/c mice.…”

Section: Introductionmentioning

confidence: 93%

Cellular immunity survey against urinary tract infection using pVAX/fimH cassette with mammalian and wild type codon usage as a DNA vaccine

Fooladi

Bagherpour

Khoramabadi

et al. 2014

Clin Exp Vaccine Res

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PurposeFimH (the adhesion fragment of type 1 fimbriae) is implicated in uropathogenic Escherichia coli (UPEC) attachment to epithelial cells through interaction with mannose. Recently, some studies have found that UPEC can thrive intracellularly causing recurrent urinary tract infection (UTI). Almost all vaccines have been designed to induce antibodies against UPEC. Yet, the humoral immune response is not potent enough to overcome neither the primary UTI nor recurrent infections. However, DNA vaccines offer the possibility of inducing cell mediated immune responses and may be a promising preventive tool.Materials and MethodsIn this study, we employed two different open reading frames within mammalian (mam) and wild type (wt) codons of fimH gene. Optimized fragments were cloned in pVAX-1. Expression of the protein in COS-7 was confirmed by western blot analysis after assessing pVAX/fimH(mam) and pVAX/fimH(wt). The constructs were injected to BALB/c mice at plantar surface of feet followed by electroporation.ResultsThe mice immunized with both constructs following booster injection with recombinant FimH showed increased interferon-γ and interleukin-12 responses significantly higher than non-immunized ones (p<0.05). The immunized mice were challenged with UPEC and then the number of bacteria recovered from the immunized mice was compared with the non-immunized ones. Decreased colony count in immunized mice along with cytokine responses confirmed the promising immune response by the DNA vaccines developed in this study.ConclusionIn conclusion, DNA vaccines of UPEC proteins may confer some levels of protection which can be improved by multiple constructs or boosters.

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“…Immunization resulted in T H 1 and T H 2 responses as assessed by cytokine responses in splenocyte proliferation assays and ratio of IgG1 to IgG2a. Lastly, another group has recently developed a mammalian codon-optimized fimH plasmid construct that could be used in a DNA vaccine, but they have not published results of trials in animals (312). …”

Section: Uti Vaccinesmentioning

confidence: 99%

Drug and Vaccine Development for the Treatment and Prevention of Urinary Tract Infections

O’Brien¹,

Hannan

Nielsen³

et al. 2016

Microbiol Spectr

112

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Urinary tract infections (UTI) are among the most common bacterial infections in humans, affecting millions of people every year. UTI cause significant morbidity in women throughout their lifespan, in infant boys, in older men, in individuals with underlying urinary tract abnormalities, and in those that require long-term urethral catheterization, such as patients with spinal cord injuries or incapacitated individuals living in nursing homes. Serious sequelae include frequent recurrences, pyelonephritis with sepsis, renal damage in young children, pre-term birth, and complications of frequent antimicrobial use including high-level antibiotic resistance and Clostridium difficile colitis. Uropathogenic E. coli (UPEC) cause the vast majority of UTI, but less common pathogens such as Enterococcus faecalis and other enterococci frequently take advantage of an abnormal or catheterized urinary tract to cause opportunistic infections. While antibiotic therapy has historically been very successful in controlling UTI, the high rate of recurrence remains a major problem, and many individuals suffer from chronically recurring UTI, requiring long-term prophylactic antibiotic regimens to prevent recurrent UTI. Furthermore, the global emergence of multi-drug resistant UPEC in the past ten years spotlights the need for alternative therapeutic and preventative strategies to combat UTI, including anti-infective drug therapies and vaccines. In this chapter, we review recent advances in the field of UTI pathogenesis, with an emphasis on the identification of promising drug and vaccine targets. We then discuss the development of new UTI drugs and vaccines, highlighting the challenges these approaches face and the need for a greater understanding of urinary tract mucosal immunity.

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Evaluation of mammalian codon usage of fimH in DNA vaccine design

Cited by 5 publications

References 40 publications

Codon Preference Optimization Increases Prokaryotic Cystatin C Expression

Codon Preference Optimization Increases Prokaryotic Cystatin C Expression

Cellular immunity survey against urinary tract infection using pVAX/fimH cassette with mammalian and wild type codon usage as a DNA vaccine

Drug and Vaccine Development for the Treatment and Prevention of Urinary Tract Infections

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