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In order to develop a safe and effective broad-spectrum vaccine for foot-and-mouth disease (FMDV), here, we developed a recombinant FMD multiple-epitope trivalent vaccine based on three distinct topotypes of FMDV. Potency of the vaccine was evaluated by immune efficacy in pigs. The results showed that the vaccine with no less than 25 μg of antigen elicited FMDV serotype O specific antibodies and neutralization antibodies by primary-booster regime, and offered immune protection to pigs. More importantly, the vaccine elicited not only the same level of neutralization antibodies against the three distinct topotypes of FMDV, but also provided complete protection in pigs from the three corresponding virus challenge. None of the fully protected pigs were able to generate anti-3ABC antibodies throughout the experiment, which implied the vaccine can offer sterilizing immunity. The vaccine elicited lasting-long high-level antibodies and effectively protected pigs from virulent challenge within six months of immunization. Therefore, we consider that this vaccine may be used in the future for the prevention and control of FMD. • Multiple-epitope vaccines is a very promising vaccine in future. • Multi-epitope vaccine can offer solid immune protection for pig against three antigenic shift isolates. • Multi-epitope vaccine can elicit broaden spectrum immune response. [ABSTRACT FROM AUTHOR]

Abstract Background shRNA targeting the integrin αv subunit, which is the foot-and-mouth disease virus (FMDV) receptor, plays a key role in virus attachment to susceptible cells. We constructed a RNAi lentiviral vector, iαv pLenti6/BLOCK -iT™, which expressed siRNA targeting the FMDV receptor, the porcine integrin αv subunit, on PK-15 cells. We also produced a lentiviral stock, established an iαv-PK-15 cell line, evaluated the gene silencing efficiency of mRNA using real-time qRT-PCR, integrand αv expression by indirect immunofluorescence assay (IIF) and cell enzyme linked immunosorbent assays (cell ELISA), and investigated the in vivo inhibitory effect of shRNA on FMDV replication in PK-15 cells. Results Our results indicated successful establishment of the iαv U6 RNAi entry vector and the iαv pLenti6/BLOCK -iT expression vector. The functional titer of obtained virus was 1.0 × 106 TU/mL. To compare with the control and mock group, the iαv-PK-15 group αv mRNA expression rate in group was reduced by 89.5%, whilst IIF and cell ELISA clearly indicated suppression in the experimental group. Thus, iαv-PK-15 cells could reduce virus growth by more than three-fold and there was a > 99% reduction in virus titer when cells were challenged with 102 TCID50 of FMDV. Conclusions Iαv-PK-15 cells were demonstrated as a cell model for anti-FMDV potency testing, and this study suggests that shRNA could be a viable therapeutic approach for controlling the severity of FMD infection and spread.

Abstract Background Foot-and-mouth disease virus (FMDV) causes an economically important and highly contagious disease of cloven-hoofed animals. RNAi triggered by small RNA molecules, including siRNAs and miRNAs, offers a new approach for controlling viral infections. There is no report available for FMDV inhibition by vector-delivered miRNA, although miRNA is believed to have more potential than siRNA. In this study, the inhibitory effects of vector-delivered miRNAs targeting the 3D gene on FMDV replication were examined. Results Four pairs of oligonucleotides encoding 3D-specific miRNA of FMDV were designed and selected for construction of miRNA expression plasmids. In the reporter assays, two of four miRNA expression plasmids were able to significantly silence the expression of 3D-GFP fusion proteins from the reporter plasmid, p3D-GFP, which was cotransfected with each miRNA expression plasmid. After detecting the silencing effects of the reporter genes, the inhibitory effects of FMDV replication were determined in the miRNA expression plasmid-transfected and FMDV-infected cells. Virus titration and real-time RT-PCR assays showed that the p3D715-miR and p3D983-miR plasmids were able to potently inhibit the replication of FMDV when BHK-21 cells were infected with FMDV. Conclusion Our results indicated that vector-delivered miRNAs targeting the 3D gene efficiently inhibits FMDV replication in vitro. This finding provides evidence that miRNAs could be used as a potential tool against FMDV infection.

Recently, many countries, including China, have experienced a series of type A and O foot‐and‐mouth disease virus (FMDV) epidemics, causing serious economic losses. Although concerns about the safety of inactivated FMD vaccines have been raised, the development of a safe and effective subunit vaccine is necessary. We constructed two chimeric virus‐like particles (VLPs; rHBc/AO and rHBc/AOT VLPs) displaying tandem repeats of B cell epitopes (VP1 residue 134‐161 and 200‐213) derived from type A and O FMDV and one T cell epitope (3 A residue 21‐35) using the truncated hepatitis B virus core (HBc) carrier. Our results indicate that the chimeric HBc can self‐assemble into VLPs with these FMDV epitopes displayed on the surface. Immunization with the chimeric VLPs induced specific IgG and neutralization antibodies against type A and O FMDV in mice. Compared with the commercial type A/O FMDV bivalent inactivated vaccine, rHBc/AO and rHBc/AOT VLPs significantly stimulated the production of Th1 type cytokines (IFN‐γ and IL‐2), whereas Th2 cytokine production (IL‐4 and IL‐10) was decreased. Compared with rHBc/AO, rHBc/AOT induced increased Th2 cytokine and specific IgG production. These results demonstrate that the VLPs constructed in the current study induced both humoral and cellular immune responses and may represent potential bivalent VLP vaccines targeting both FMDV type A and O strains. [ABSTRACT FROM AUTHOR]

Chemiluminescence technique as a novel detection method has gained much attention in recent years owning to the merits of high sensitivity, wider linear ranges, and low background signal. Similarly, nanotechnology especially for gold nanoparticles has emerged as detection tools due to their unique physical and chemical properties. Recently, it has become increasingly popular to couple gold nanoparticles with chemiluminescence technique in biological agents' detection. In this review, we describe the superiority of both chemiluminescence and gold nanoparticles and conclude the different applications of gold nanoparticle-initialed chemiluminescence in biomedical detection. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease virus (FMDV) serotype Asia 1 was mostly endemic in Asia and then was responsible for economically important viral disease of cloven-hoofed animals, but the study on its selection and evolutionary process is comparatively rare. In this study, we characterized 377 isolates from Asia collected up until 2012, including four vaccine strains. Maximum likelihood analysis suggested that the strains circulating in Asia were classified into 8 different groups (groups I–VIII) or were unclassified (viruses collected before 2000). On the basis of divergence time analyses, we infer that the TMRCA of Asia 1 virus existed approximately 86.29 years ago. The result suggested that the virus had a high mutation rate (5.745 × 10−3 substitutions/site/year) in comparison to the other serotypes of FMDV VP1 gene. Furthermore, the structural protein VP1 was under lower selection pressure and the positive selection occurred at many sites, and four codons (positions 141, 146, 151, and 169) were located in known critical antigenic residues. The remaining sites were not located in known functional regions and were moderately conserved, and the reason for supporting all sites under positive selection remains to be elucidated because the power of these analyses was largely unknown. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD), caused by foot-and-mouth disease virus (FMDV), is considered one of the most important viral diseases of cloven-hoofed animals, causing severe economic losses in affected regions of the world. Three serotypes (A, O, and Asia 1) of FMDV have been identified in China since 1958. In addition, the occurrence of novel subtypes within these serotypes has made the epidemiology of FMDV more complicated over the last few years. In this review, we summarize the history and the current epidemiological situation in China, genetic diversity (e.g., quasispecies dynamics, antigenic heterogeneity, and functional constraints), intertypic recombination, and the evidence for positive selection of different FMDV serotypes. We also assess these genetic data to understand the origin, evolution, and transmission of FMDV, the findings of which may be useful in developing control measures for future epidemics. [ABSTRACT FROM AUTHOR]

A bovine viral diarrhea disease virus (BVDV) GS-4 was isolated in Western China form dairy cattle with respiratory disease. Genomic comparison analysis with the 5′ half genome sequence encompassing the coding region of N, capsid, and envelope glycoproteins showed that the GS-4 should be classified into BVDV-1b1, which is considered as one of the predominant subgenotypes found in China. This classification was confirmed by phylogenetic analysis based on E2 coding region. [ABSTRACT FROM AUTHOR]

Abstract: Bovine viral diarrhea virus (BVDV) infects both domestic and wild animals, causing substantial economic losses. In order to investigate possible infection in Bactrian camels in Western China, a total of 56 blood samples were collected from clinically healthy Bactrian camels and tested for BVDV antigens and antibodies using antigen capture enzyme-linked immunosorbent assay (ELISA) and virus neutralization test. The antigen-positive samples (n =17) were further tested for viral nucleic acids by species-specific real-time RT-PCR assays, which showed presence of BVDV-1, but not BVDV-2 nor atypical bovine pestivirus, in the camel samples. Twelve non-cytopathogenic viruses were isolated and genetically typed by sequencing of the 5′untranslated region (5′UTR) and Npro coding sequences. Phylogenetic analysis divided the isolates into six known subgenotypes: BVDV-1a, BVDV-1b, BVDV-1c, BVDV-1m, BVDV-1o, BVDV-1p and a putative subgenotype, BVDV-1q. This study provides, for the first time, serological and molecular evidence for natural infection of Bactrian camels in Western China with highly divergent BVDV-1 strains. Further investigations are needed to elucidate the possible roles of Bactrian camels in the epidemiology of BVD in Western China. [Copyright &y& Elsevier]

• ASFV is one of the "notifiable diseases" listed by the OIE. • Urgency of developing effective vaccines to combat the devastating viral disease in pigs. • Enhancement in antigen-specific cellular and humoral immunity induced by ASFV proteins. • Providing valuable information to develop subunit vaccines against ASF. African swine fever (ASF) is a highly fatal swine disease threatening the global pig industry. Currently, vaccine is not commercially available for ASF. Hence, it is desirable to develop effective subunit vaccines against ASF. Here, we expressed and purified two recombinant fusion proteins comprising ASFV proteins p30 and p54 fused to a novel cell-penetrating peptide Z12, which were labeled as ZPM (Z12-p30-modified p54) and ZPMT (Z12-p30-modified p54-T cell epitope). Purified recombinant p30 and modified p54 expressed alone or fused served as controls. The transduction capacity of these recombinant proteins was assessed in RAW264.7 cells. Both ZPM and ZPMT exhibited higher transduction efficiency than the other proteins. Subsequently, humoral and cellular immune responses elicited by these proteins were evaluated in mice. ZPMT elicited the highest levels of antigen-specific IgG responses, cytokines (interleukin-2, interferon-γ, and tumor necrosis factor-α) and lymphocyte proliferation. Importantly, sera from mice immunized with ZPM or ZPMT neutralized greater than 85% of ASFV in vitro. Our results indicate that ZPMT induces potent neutralizing antibody responses and cellular immunity in mice. Therefore, ZPMT may be a suitable candidate to elicit immune responses in swine, providing valuable information for the development of subunit vaccines against ASF. [ABSTRACT FROM AUTHOR]