A Multiepitope Fusion Antigen Elicits Neutralizing Antibodies against Enterotoxigenic Escherichia coli and Homologous Bovine Viral Diarrhea Virus<i>In Vitro</i>

Hashish, Emad A.; Zhang, Chengxian; Ruan, Xiaosai; Knudsen, David; Chase, Christopher; Isaacson, Richard E.; Zhou, Guoqiang; Zhang, Weiping

doi:10.1128/cvi.00249-13

Cited by 11 publications

(9 citation statements)

References 39 publications

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“…An impediment to creating better vaccines for weaning diarrhea is that unlike neonatal diarrhea, good and reproducible pig disease models are not available. A newer approach that has been developed is based on the creation of vaccines that incorporate the expected protective epitopes of fimbriae and enterotoxins in a single protein molecule (548–551). The incorporation of epitopes from F4, F18, and heat stable and heat labile enterotoxins in a single protein molecule has been shown to elicit antibodies that can neutralize attachment of ETEC to mucosal surfaces and neutralize the enterotoxins.…”

Section: Vaccines and Colonization Inhibitorsmentioning

confidence: 99%

Animal Enterotoxigenic Escherichia coli

2016

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Enterotoxigenic Escherichia coli (ETEC) is the most common cause of E. coli diarrhea in farm animals. ETEC are characterized by the ability to produce two types of virulence factors; adhesins that promote binding to specific enterocyte receptors for intestinal colonization and enterotoxins responsible for fluid secretion. The best-characterized adhesins are expressed in the context of fimbriae, such as the F4 (also designated K88), F5 (K99), F6 (987P), F17 and F18 fimbriae. Once established in the animal small intestine, ETEC produces enterotoxin(s) that lead to diarrhea. The enterotoxins belong to two major classes; heat-labile toxin that consist of one active and five binding subunits (LT), and heat-stable toxins that are small polypeptides (STa, STb, and EAST1). This chapter describes the disease and pathogenesis of animal ETEC, the corresponding virulence genes and protein products of these bacteria, their regulation and targets in animal hosts, as well as mechanisms of action. Furthermore, vaccines, inhibitors, probiotics and the identification of potential new targets identified by genomics are presented in the context of animal ETEC.

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Section: Vaccines and Colonization Inhibitorsmentioning

confidence: 99%

Animal Enterotoxigenic Escherichia coli

2016

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“…Bovine rotavirus infection in young cattle less than 6 months old may not show typical symptoms as of BVDV. Both BRV and BVDV are frequently exhibits acute watery diarrhea and emaciation [ 12 , 13 ]. Therefore, it is important to diagnose and differentiate these infectious viruses accurately for the rapid control and prevention strategies [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Development of a GeXP-multiplex PCR assay for the simultaneous detection and differentiation of six cattle viruses

Fan

Xie

et al. 2017

PLoS ONE

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Foot-and-mouth disease virus (FMDV), Bluetongue virus (BTV), Vesicular stomatitis Virus (VSV), Bovine viral diarrheal (BVDV), Bovine rotavirus (BRV), and Bovine herpesvirus 1 (IBRV) are common cattle infectious viruses that cause a great economic loss every year in many parts of the world. A rapid and high-throughput GenomeLab Gene Expression Profiler (GeXP) analyzer-based multiplex PCR assay was developed for the simultaneous detection and differentiation of these six cattle viruses. Six pairs of chimeric primers consisting of both the gene-specific primer and a universal primer were designed and used for amplification. Then capillary electrophoresis was used to separate the fluorescent labeled PCR products according to the amplicons size. The specificity of GeXP-multiplex PCR assay was examined with samples of the single template and mixed template of six viruses. The sensitivity was evaluated using the GeXP-multiplex PCR assay on serial 10-fold dilutions of ssRNAs obtained via in vitro transcription. To further evaluate the reliability, 305 clinical samples were tested by the GeXP-multiplex PCR assay. The results showed that the corresponding virus specific fragments of genes were amplified. The detection limit of the GeXP-multiplex PCR assay was 100 copies/μL in a mixed sample of ssRNAs containing target genes of six different cattle viruses, whereas the detection limit for the Gexp-mono PCR assay for a single target gene was 10 copies/μL. In detection of viruses in 305 clinical samples, the results of GeXP were consistent with simplex real-time PCR. Analysis of positive samples by sequencing demonstrated that the GeXP-multiplex PCR assay had no false positive samples of nonspecific amplification. In conclusion, this GeXP-multiplex PCR assay is a high throughput, specific, sensitive, rapid and simple method for the detection and differentiation of six cattle viruses. It is an effective tool that can be applied for the rapid differential diagnosis of clinical samples and for epidemiological investigation.

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“…FanC‐STa‐E2 fusion protein was well expressed by IPTG‐induction (1 mM) at 37°C for 4 hr and purified by Ni‐NTA agarose using His‐tag. This multiepitope fusion protein could develop neutralizing antibodies against ETEC and BVDV in vitro [Hashish et al, ].…”

Section: Discussionmentioning

confidence: 99%

Design and production of a multiepitope construct derived fromhepatitis E viruscapsid protein

2015

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The aim of this study was to design a high density multiepitope protein, which can be a promising multiepitope vaccine candidate against Hepatitis E virus (HEV). Initially, conserved and antigenic helper T-lymphocyte (HTL) epitopes in the HEV capsid protein were predicted by in silico analysis. Subsequently, a multiepitope comprising four HTL epitopes with high-affinity binding to the HLA molecules was designed, and repeated four times as high density multiepitope construct. This construct was synthesized and cloned into pET-30a (+) vector. Then, it was transformed and expressed in Escherichia coli BL21 cells. The high density multiepitope protein was purified by Ni-NTA agarose and concentrated using Amicon filters. Finally, the immunological properties of this high density multiepitope protein were evaluated in vitro. The results showed that the high density multiepitope construct was successfully expressed and purified. SDS-PAGE and Western blot analyses showed the presence of a high density multiepitope protein band of approximately 33 kDa. Approximately 1 mg of the purified protein was obtained from each liter of the culture media. Moreover, the purified multiepitope protein was capable of induction of proliferation responses, IFN-γ ELISPOT responses and IFN-γ and IL-12 cytokines production in a significant level in peripheral blood mononuclear cells (PBMCs) isolated from HEV-recovered individuals compared to the control group. In conclusion, the newly produced multiepitope protein can induce significant T helper type 1 responses in vitro, and can be considered as a novel strategy for the development of HEV vaccines in the future.

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A Multiepitope Fusion Antigen Elicits Neutralizing Antibodies against Enterotoxigenic Escherichia coli and Homologous Bovine Viral Diarrhea VirusIn Vitro

Cited by 11 publications

References 39 publications

Animal Enterotoxigenic Escherichia coli

Animal Enterotoxigenic Escherichia coli

Development of a GeXP-multiplex PCR assay for the simultaneous detection and differentiation of six cattle viruses

Design and production of a multiepitope construct derived fromhepatitis E viruscapsid protein

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