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In the present study, we report the sequences and characterization of the porcine integrin cDNAs encoding alphav, beta3, beta6 and beta8 subunits and compare them to those of other species. The coding sequences for the porcine alphav, beta3, beta6 and beta8 subunits were found to be 3141, 2289, 2367 and 2304 nucleotides in length, encoding 1046, 762, 788 and 767-amino-acid-residue protein, respectively. The porcine integrin alphav, beta3, beta6 and beta8 subunit shares common structural and functional elements with their counterparts from the other species. Phylogenetic trees showed that the porcine alphav, beta3, beta6 and beta8 were clustered into the Artiodactyla group, together with those of camels, sheep, and cattle, that are susceptible to FMDV infection. Real-time RT-PCR was used to investigate expression of the integrins alphavbeta3, alphavbeta6 and alphavbeta8 in different tissues of pigs in order to determine the role of these receptors in tissue tropism. Expression analysis showed that alphavbeta6 and alphavbeta8 mRNA expression were detected at high levels in tissues known to support replication of FMDV. Tissue distribution pattern of alphavbeta3 mRNA seems to be unrelated to the known tissue tropism of FMDV. This study provided the first data of porcine integrins for the further studies of the FMDV pathogenesis in pigs., (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)

Integrins are heterodimeric adhesion receptors that participate in a variety of cell-cell and cell-extracellular matrix protein interactions. Many integrins recognize RGD sequences displayed on extracellular matrix proteins and the exposed loops of viral capsid proteins. Four members of the alphav integrin family of cellular receptors, alphavbeta3, alphavbeta6, alphavbeta1 and alphavbeta8, have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro, and integrins are believed to be the receptors used to target epithelial cells in the infected animals. To analyse the roles of the alphav integrins from a susceptible species as viral receptors, we have cloned Bactrian camel alphav, beta3 and beta6 integrin cDNAs and compared them to those of other species. The coding sequences for Bactrian camel integrin alphav, beta3 and beta6 were found to be 3165, 2289 and 2367 nucleotides in length, encoding 1054, 762 and 788 amino acids, respectively. The Bactrian camel alphav, beta3 and beta6 subunits share many structural features with homologues of other species, including the ligand binding domain and cysteine-rich region. Phylogenetic trees and similarity analyses showed the close relationships of integrin genes from Bactrian camels, pigs and cattle, which are each susceptible to FMDV infection, that were distinct from the orders Rodentia, Primates, Perissodactyla, Carnivora, Galliformes and Xenopus. We postulate that host tropism of FMDV may in part be related to the divergence in integrin subunits among different species.

Four members of the alphav integrin family of cellular receptors, alphavbeta1, alphavbeta3, alphavbeta6, and alphavbeta8, have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro, and integrins are believed to be the receptors used to target epithelial cells in the infected animals. To analyse roles of the alphav integrins from a susceptible species as viral receptors, we have cloned sheep alphav, beta1, and beta6 integrin cDNAs and compared them to those of other species. The coding sequences for sheep integrin alphav, beta1, and beta6 were found to be 3147, 2397, and 2364 nuclotides in length, encoding 1048, 798, and 787 amino acids, respectively. The sheep alphav, beta1, and beta6 subunits share many structural features including ligand binding domain and cysteine-rich region with homologues of other species. Phylogenetic trees and similarity analyses showed the close relationship of integrin genes among sheep, pigs, cattle and Bactrian camels that are susceptible to FMDV infection, which were distinct from the order Rodentia, Primates, Perissodactyla, Carnivora, Galliformes. We postulate that host tropism of FMDV may be related to divergence in integrin subunits among different species.

Foot-and-mouth disease virus (FMDV) is an RNA virus that causes a highly contagious disease in domestic and wild cloven-hoofed animals. Although vaccination has been used to protect animals against FMDV, there are shortcomings in the efficacy of the available vaccines. RNA interference (RNAi) is triggered by small RNA molecules, including short interfering RNAs and microRNAs (miRNAs), and the use of RNAi-based methods have demonstrated promise as an alternative method of controlling the transmission of FMDV. However, the method of delivery, short duration of siRNA and miRNA in vivo , and the genetic variability of FMDV confound the use of RNAi-based strategies for FMDV control. FMDV has been shown to exploit host-cell integrins as cell-surface receptors to initiate infection. We selected the gene for the integrin αv subunit as an RNAi target, and constructed three αv-specific miRNA expression plasmids. The effects of these miRNAs on FMDV infection were examined in PK-15 cells and transgenic suckling mice. In PK-15 cells, the expression of the αv-specific miRNAs significantly inhibited the expression of integrin αv receptor and decreased FMDV infection. The transgenic mice were generated by integrating the αv-specific miRNA expression cassette using pronuclear microinjection. When challenged with a dose of FMDV ten times greater than the LD 50 , the survival rate of transgenic suckling mice was approximately six-fold higher than that of their non-transgenic littermates, indicating that the interference of the miRNAs significantly reduced FMDV infection in the transgenic mice. This is the first report of limiting FMDV attachment to cellular receptors using miRNA-mediated gene knock down of cell-surface receptors to significantly reduce FMDV infection in cell culture and transgenic suckling mice. [ABSTRACT FROM AUTHOR]