Computational prediction of MHC class I epitopes for most common viral diseases in cattle (Bos taurus).

Viral diseases like foot-and-mouth disease (FMD), calf scour (CS), bovine viral diarrhea (BVD), infectious bovine rhinotracheitis (IBR) etc. affect the growth and milk production of cattle (Bos taurus) causing severe economic loss. Epitope-based vaccine designing have been evolved to provide a new strategy for therapeutic application of pathogen-specific immunity in animals. Therefore, identification of major histocompatibility complex (MHC) binding peptides as potential T-cell epitopes is widely applied in peptide vaccine designing and immunotherapy. In this study, MetaMHCI tool was used with seven different algorithms to predict the potential T-cell epitopes for FMD, BVD, IBR and CS in cattle. A total of 54 protein sequences were filtered out from a total set of 6351 sequences of the pathogens causing the said diseases using bioinformatics approaches. These selected protein sequences were used as the key inputs for MetaMHCI tool to predict the epitopes for the BoLA-A11 MHC class I allele of B. taurus. Further, the epitopes were ranked based on a proposed principal component analysis based epitope score (PbES). The best epitope for each disease based on its predictability through maximum number of predictors and low PbES was modeled in PEP-FOLD server and docked with the BoLA-A11 protein for understanding the MHC-epitope interaction. Finally, a total of 78 epitopes were predicted, out of which 27 were for FMD, 25 for BVD, 12 for CS and 14 for IBR. These epitopes could be artificially synthesized and recommended to vaccinate the cattle for the considered diseases. Besides, the methodology adapted here could also be used to predict and analyze the epitopes for other microbial diseases of important animal species.

Comparative studies of the potency of foot and mouth disease virus trivalent vaccine with different concentration of the antigenic content [146S]

Estimation of antigenic content [146S] of FMDV serotypes [A, O, SAT2] by sucrose density gradient [SDG] ultracentrifugation by determining the absorbance at 254 nm using ISCO520C density gradient system to produce a highly potent trivalent virus vaccine. The antigenic mass 146S [microg/ml] of serotype [O Pan Asia2, A Iran O5 and SAT2/EGY/2012] were 6.5, 6.2 and 5.9, respectively. The vaccine was injected into three groups of calves [2individuals/each group] subcutaneously in lateral part of the neck for a dose 3 ml [6.2 microg/serotype/ml], a dose 1.5 ml [4.1 microg/serotype/ml] and a dose 1 ml [2 micro/ml], the sera samples were collected at 7[th] day post vaccination [dpv], 14[th] dpv, 21[th] dpv, 28[th] dpv and every 2 wks till 40 weeks to evaluate the immune response along that period. The antibody titers/40wpv for a 3 ml dose [6.2 microg/ml] of serotypes [O Pan Asia-2, A Iran O5 and SAT-2/EGY/2012] were 2.08, 2 and 1.94, respectively [over the protective titer, PT=1.5 in SNT for cattle], a dose [4.1 microg/ml] of the three serotypes were 1.56, 1.62 and 1.63 [over PT], respectively, but for [2 microg/ml] dose of the three serotypes, the antibodies titer were 1.25, 1.19 and 1.2 [below PT], that show the antibodies titer depend on the concentration of the antigenic mass [146S] and with increase of the 146S concentration increase of the potency of the vaccine. The potency testing of the study depend upon the correlation between 146S and the neutralizing antibody titers were measured by SNT which are the perfect alternative of other potency tests which employ the challenge of the cattle with virulent virus. The immune response of the highly potent vaccine [4.1 microg/serotype/ml and 6.2 microg/serotype/ml] started early after 1[st] wpv and the protective titer remain for more than 38 wpv [especially in 6.2 microg/ml injected calves] and that confer the potency of the vaccine of that dose

Establishment and evaluation of a murine alphavbeta3-integrin-expressing cell line with increased susceptibility to Foot-and-mouth disease virus

Integrin alphavbeta3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and roles of alphavbeta3 integrin, a stable CHO-677 cell line expressing the murine alphavbeta3 heterodimer (designated as "CHO-677-malphavbeta3" cells) was established using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits alphav and beta3 were detected at the gene and protein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-malphavbeta3 cell line at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-forming assay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replication levels of Foot-and-mouth disease virus (FMDV) in the CHO-677-malphavbeta3 cell line. After infection with FMDV/O/ZK/93, the cell line showed a significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stable alphavbeta3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of alphavbeta3 integrin, and as a cell model for FMDV research.

[Comparative study on 1D [VP1] region of foot and mouth disease virus [type A strain] among different isolates: Khorasan Razavi isolate and other Iranian and neighboring countries isolates]

Foot-and-mouth disease [FMD] is one of the most important virus disease in farm animals. Types O, A and Asial FMD virus have been endemic in Iran. In this study, samples from suspected livestock were analyzed by RT-PCR experiment. The number of 702 nucleotides determined at 1D- 2B region of type A strain isolated from Khorasan Razavi province sequenced and compared with that of other reported isolates type A from Iran and neighboring countries. The results show that field isolated type A has about 89% similarity with other reported isolates type A from Iran and neighboring countries. Furthermore, this virus shows the most similarity with A/IRN/1/87[Samuel. Phylogenitic analysis revealed that virus was closely related to A22-Iraq/99 and A/IRN/iso/105 that rest in the same lineage. The data showed high similarity between type A viruses involved in the Khorasan Razavi province and A/IRN/87v [vaccine strain]; so that it can be concluded that the vaccine can produce prophylactic antibody against this virus