Article

# Genetic characterization of the cell-adapted PanAsia strain of foot-and-mouth disease virus O/Fujian/CHA/5/99 isolated from swine

[more]
National Foot-and-Mouth Disease Reference Laboratory, State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of the Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu 730046, PR China.
(Impact Factor: 2.18). 08/2010; 7(1):208. DOI: 10.1186/1743-422X-7-208
Source: PubMed

ABSTRACT

According to Office International Des Epizooties (OIE) Bulletin, the PanAsia strain of Foot-and-Mouth Disease Virus (FMDV) was invaded into the People's Republic of China in May 1999. It was confirmed that the outbreaks occurred in Tibet, Hainan and Fujian provinces. In total, 1280 susceptible animals (68 cattle, 1212 swine) were destroyed for the epidemic control.To investigate the distinct biological properties, we performed plaque assay, estimated the pathogenicity in suckling mice and determined the complete genomic sequence of FMDV swine-isolated O/Fujian/CHA/5/99 strain. In addition, a molecular modeling was carried out with the external capsid proteins.
The pathogenicity study showed that O/Fujian/CHA/5/99 had high virulence with respect to infection in 3-day-old suckling-mice (LD50 = 10-8.3), compared to O/Tibet/CHA/1/99 (LD50 = 10-7.0) which isolated from bovine. The plaque assay was distinguishable between O/Fujian/CHA/5/99 and O/Tibet/CHA/1/99 by their plaque phenotypes. O/Fujian/CHA/5/99 formed large plaque while O/Tibet/CHA/1/99 formed small plaque.The 8,172 nucleotides (nt) of O/Fujian/CHA/5/99 was sequenced, and a phylogenetic tree was generated from the complete nucleotide sequences of VP1 compared with other FMDV reference strains. The identity data showed that O/Fujian/CHA/5/99 is closely related to O/AS/SKR/2002 (94.1% similarity). Based on multiple sequence alignments, comparison of sequences showed that the characteristic nucleotide/amino acid mutations were found in the whole genome of O/Fujian/CHA/5/99.
Our finding suggested that C275T substitution in IRES of O/Fujian/CHA/5/99 may induce the stability of domain 3 for the whole element function. The structure prediction indicated that most of 14 amino acid substitutions are fixed in the capsid of O/Fujian/CHA/5/99 around B-C loop and E-F loop of VP2 (antigenic site 2), and G-H loop of VP1 (antigenic site 1), respectively. These results implicated that these substitutions close to heparin binding sites (E136G in VP2, A174 S in VP3) and at antigenic site 1 (T142A, A152T and Q153P in VP1) may influence plaque size and the pathogenicity to suckling mice.The potential of genetic characterization would be useful for microevolution and viral pathogenesis of FMDV in the further study.

### Full-text

Available from: Zaixin Liu, Oct 07, 2015
0 Followers
·
22 Reads
• Source
• "BHK-21, WT-CHO and pgsD-677 cell monolayers in 6-well plates were inoculated with the serial ten-fold dilutions of all the selected PanAsia-1 strains and rescued viruses, following the previously described procedures [46]. The values of plaque titrations of each virus were calculated as described previously [24]. "
##### Article: Effects of two amino acid substitutions in the capsid proteins on the interaction of two cell-adapted PanAsia-1 strains of foot-and-mouth disease virus serotype O with heparan sulfate receptor
[Hide abstract]
ABSTRACT: Background Some cell-adapted strains of foot-and-mouth disease virus (FMDV) can utilize heparan sulfate (HS) as a receptor to facilitate viral infection in cultured cells. A number of independent sites on the capsid that might be involved in FMDV-HS interaction have been studied. However, the previously reported residues do not adequately explain HS-dependent infection of two cell-adapted PanAsia-1 strains (O/Tibet/CHA/6/99tc and O/Fujian/CHA/9/99tc) of FMDV serotype O. To identify the molecular determinant(s) for the interaction of O/Tibet/CHA/6/99tc and O/Fujian/CHA/9/99tc with HS receptor, several chimeric viruses and site-directed mutants were generated by using an infectious cDNA of a non-HS-utilizing rescued virus (Cathay topotype) as the genomic backbone. Phenotypic properties of these viruses were determined by plaque assays and virus adsorption and penetration assays in cultured cells. Results Only two of the rescued viruses encoding VP0 of O/Tibet/CHA/6/99tc or VP1 of O/Fujian/CHA/9/99tc formed plaques on wild-type Chinese hamster ovary (WT-CHO; HS+) cells, but not on HS-negative pgsD-677 cells. The formation of plaques by these two chimeric viruses on WT-CHO cells could be abolished by the introduction of single amino acid mutations Gln-2080 → Leu in VP2 of O/Tibet/CHA/6/99tc and Lys-1083 → Glu in VP1 of O/Fujian/CHA/9/99tc, respectively. Nonetheless, the introduced mutation Leu-2080 → Gln in VP2 of O/Fujian/CHA/9/99tc for the construction of expectant recombinant plasmid led to non-infectious progeny virus in baby hamster kidney 21 (BHK-21) cells, and the site-directed mutant encoding Glu-1083 → Lys in VP1 of O/Tibet/CHA/6/99tc did not acquire the ability to produce plaques on WT-CHO cells. Significant differences in the inhibition of the infectivity of four HS-utilizing viruses by heparin and RGD-containing peptide were observed in BHK-21 cells. Interestingly, the chimeric virus encoding VP0 of O/Fujian/CHA/9/99tc, and the site-directed mutant encoding Gln-2080 → Leu in VP2 of O/Tibet/CHA/6/99tc could bind to HS, but there was no expression of the 3A protein of these two viruses in WT-CHO cells. Conclusion The results suggest that the cooperation of certain specific amino acid residues in the capsid proteins of these two cell-adapted PanAsia-1 strains is essential for viral infectivity, the heparin affinity and the capability on FMDV-HS interaction.
Virology Journal 07/2014; 11(1):132. DOI:10.1186/1743-422X-11-132 · 2.18 Impact Factor
• Source
• "Genomic sequencing of the structural proteins revealed two aa substitutions (VP2 D133N and VP3 H56R) in the mildly pathogenic small plaque virus when compared to the large plaque virus in suckling mice (Kanno et al., 2002; Morioka et al., 2008). Nevertheless, these two mutations did not account for the characteristics of another PanAsia strain isolated from China (Bai et al., 2010). This study demonstrated that the BHK-21 cell-adapted O/Fujian/CHA/5/99 strain had high virulence in suckling mice and formed larger plaques than the bovine-isolated O/Tibet/CHA/1/99, while sequence analysis revealed that both strains carried a D residue in position 133 of VP2 and an H in position 56 of VP3. "
##### Article: Role of arginine-56 within the structural protein VP3 of foot-and-mouth disease virus (FMDV) O1 Campos in virus virulence
[Hide abstract]
ABSTRACT: FMDV O1 subtype undergoes antigenic variation under diverse growth conditions. Of particular interest is the amino acid variation observed at position 56 within the structural protein VP3. Selective pressures influence whether histidine (H) or arginine (R) is present at this position, ultimately influencing in vitro plaque morphology and in vivo pathogenesis in cattle. Using reverse genetics techniques, we have constructed FMDV type O1 Campos variants differing only at VP3 position 56, possessing either an H or R (O1Ca-VP3-56H and O1Ca-VP3-56R, respectively), and characterized their in vitro phenotype and virulence in the natural host. Both viruses showed similar growth kinetics in vitro. Conversely, they had distinct temperature-sensitivity (ts) and displayed significantly different pathogenic profiles in cattle and swine. O1Ca-VP3-56H was thermo stable and induced typical clinical signs of FMD, whereas O1Ca-VP3-56R presented a ts phenotype and was nonpathogenic unless VP3 position 56 reverted in vivo to either H or cysteine (C).
Virology 01/2012; 422(1):37-45. DOI:10.1016/j.virol.2011.09.031 · 3.32 Impact Factor
• Source
##### Article: A compactness result for Landau state in thin-film micromagnetics
[Hide abstract]
ABSTRACT: We deal with a nonconvex and nonlocal variational problem coming from thin-film micromagnetics. It consists in a free-energy functional depending on two small parameters $\eps$ and $\eta$ and defined over $S^2-$vector fields $m$ that are tangent at the boundary of a two-dimensional domain $\Omega$. We are interested in the behavior of minimizers as $\eps, \eta \to 0$. The minimizers tend to be in-plane away from a region of length scale $\eps$ (generically, an interior vortex ball or two boundary vortex balls) and of vanishing divergence, so that $S^1-$transition layers of length scale $\eta$ (N\'eel walls) are enforced by the boundary condition. We first prove an upper bound for the minimal energy that corresponds to the cost of a vortex and the configuration of N\'eel walls associated to the viscosity solution, so-called Landau state. Our main result concerns the compactness of vector fields $m_{\eps, \eta}$ of energies close to the Landau state in the regime where a vortex is energetically more expensive than a N\'eel wall. Our method uses techniques developed for the Ginzburg-Landau type problems for the concentration of energy on vortex balls, together with an approximation argument of $S^2-$vector fields by $S^1-$vector fields away from the vortex balls.
09/2010; 28(2). DOI:10.1016/j.anihpc.2011.01.001
Show more