Transcriptional profiling of feline infectious peritonitis virus infection in CRFK cells and in PBMCs from FIP diagnosed catsReport as inadecuate




Transcriptional profiling of feline infectious peritonitis virus infection in CRFK cells and in PBMCs from FIP diagnosed cats - Download this document for free, or read online. Document in PDF available to download.

Virology Journal

, 10:329

Veterinary DNA viruses

Abstract

BackgroundFeline Infectious Peritonitis FIP is a lethal systemic disease, caused by the FIP Virus FIPV; a virulent mutant of Feline Enteric Coronavirus FECV. Currently, the viruses virulence determinants and host gene expressions during FIPV infection are not fully understood.

MethodsRNA sequencing of Crandell Rees Feline Kidney CRFK cells, infected with FIPV strain 79–1146 at 3 hours post infection h.p.i, were sequenced using the Illumina next generation sequencing approach. Bioinformatic’s analysis, based on Felis catus 2X annotated shotgun reference genome, using CLC bio Genome Workbench mapped both control and infected cell reads to 18899 genes out of 19046 annotated genes. Kal’s Z test statistical analysis was used to analyse the differentially expressed genes from the infected CRFK cells. Real time RT-qPCR was developed for further transcriptional profiling of three genes PD-1, PD-L1 and A3H in infected CRFK cells and Peripheral Blood Mononuclear Cells PBMCs from healthy and FIP-diseased cats.

ResultsBased on Kal’s Z-test, with False Discovery Rate FDR <0.05 and >1.99 fold change on gene expressions, a total of 61 genes were differentially expressed by both samples, where 44 genes were up-regulated and the remainder were down-regulated. Most genes were closely clustered together, suggesting a homogeneous expression. The majority of the genes that were significantly regulated, were those associated with monocytes-macrophage and Th1 cell functions, and the regulation of apoptosis. Real time RT-qPCR developed focusing on 2 up-regulated genes PD-L1 and A3H together with an apoptosis associated gene PD-1 expressions in FIPV infected CRFK cells and in PBMCs from healthy and FIP diagnosed cats produced concordant results with transcriptome data.

ConclusionThe possible roles of these genes, and their importance in feline coronaviruses infection, are discussed.

KeywordsFIPV CRFK PBMCs Transcriptome RT-qPCR Gene expression Fold change AbbreviationsBLASTBasic Local Alignment Search Tool

CRFKCrandell Rees Feline Kidney

CMICell-mediated immunity

FCOVFeline coronavirus

FDRFalse discovery rate

FECVFeline enteric coronavirus

FIPVFeline infectious peritonitis virus

FIVFeline immunodeficiency virus

kbKilobase

minMinute

mlMililitre

MOIMultiplicity of infection

NGSNext generation sequencing

PBMCsPeripheral Blood Mononuclear Cells

PCRPolymerase chain reaction

RPKMReads per kilobase of exon model per million mapped reads.

Electronic supplementary materialThe online version of this article doi:10.1186-1743-422X-10-329 contains supplementary material, which is available to authorized users.

Download fulltext PDF



Author: Mohammad Syamsul Reza Harun - Choong Oi Kuan - Gayathri Thevi Selvarajah - Tan Sheau Wei - Siti Suri Arshad - Mohd Hai

Source: https://link.springer.com/







Related documents