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Unravelling the cellular and humoral immune responses of Antheraea pernyi hemolymph to Antheraea pernyi nucleopolyhedrovirus infection

ABSTRACT: Purpose: With the advent of next generation sequencing technologies, several researches have been focused on the molecular mechanism underlying the innate immunity of insects based on the changes of gene expression profiles in hemolymph. The goal of this study is to detect the differences in transcript levels in A. pernyi hemolymph in response to ApNPV. Methods:The A. pernyi hemolymph infected by ApNPV and normal samples were generated by deep sequencing, in triplicate, using Illumina Hiseq 4000. The high-quality reads were obtained by removing the reads that contained adaptor contamination, low quality bases and undetermined bases.The transcriptome were de novo assembly. Results:We constructed six cDNA libraries using RNA samples from the hemolymph in ApNPV-infected and non-infected A. pernyi larvae with three replicates for each group. A total of 50459962, 49797542, 54402474 and 56342454, 56036432, 55471402 raw reads in ApNPV-infected and control groups were generated. After filtering, 48009732, 47241848, 51319586 and 53555764, 53364108, 53077540 clean reads were obtained. All short-read sequences were assembled into 49966 transcripts and 37498 unigenes. Conclusions: The transcriptome sequences were de novo assembled. The differentially expressed genes (DEGs) including both up- and down-regulated genes were obtained. We focused on the innate immunity related DEGs, and screened numerous of DEGs involved in cellular and humoral immune related pathways via bioinformatic analysis. The results of this study lay the foundation for a better understanding of the molecular mechanisms of cellular and humoral immunity of A. pernyi.

ORGANISM(S): Antheraea pernyi

PROVIDER: GSE125800 | GEO | 2019/01/29

REPOSITORIES: GEO

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Project description:Purpose: The goals of this study are to investigate the differentially expressed genes between ALV-J infected (WRR+) and uninfected (WRR-)chickens spleens by Illumina deep sequencing. Methods: 140-day-old female chickens of White Recessive Rock (WRR) were confirmed as J subgroup avian leukosis virus (ALV-J) infection. Total RNA from three ALV-J-infected spleens (designated: WRR1+, WRR2+, WRR3+) and three uninfected normal spleen samples (designated: WRR1-, WRR2-, WRR3-) was isolated by TRIzol following the manufacturer’s instruction (Invitrogen, CA, USA). RNA samples of three individuals within each group were pooled with equal amounts, and then were subjected to Illumina deep sequencing by Illumina Genome Analyzer IIx. Results: Through raw data processed, 49,979,648 and 43,704,401 clean reads with an average length of 101 bp, which represented total residues of 4,859,084,087 and 4,238,826,168 bp, were obtained for WRR- and WRR+ libraries, respectively. Subsequently, the clean reads in the two libraries were assembled. Altogether, 121,493 contigs were assembled with an average length of 927 bp (ranged from 300 bp to 23,402 bp), leading to generation of 82,829 unigenes. The length of unigenes varied from 351 bp to 28,928 bp, with an average length of 1,155 bp. Based on the FPKM value of each gene, 252 DEGs were identified by DEGseq package using Benjamini-q-value of 0.05 as a cut-off. In ALV-J infected spleens, 90 genes showed up-regulated and 162 showed down-regulated expression when compared to uninfected samples. Conclusions: Our study represents the first time to elucidate the ALV-J infected chickens’spleens at the transcription level by RNA-seq technology. A total of 252 genes were found to be differentially expressed in ALV-J infected spleens when compared to uninfected chickens. These genes can be considered as candidates for further study ALV-J invasion. Spleen mRNA profiles of 140-day-old ALV-J infected (WRR+) and uninfected (WRR-) female chickens of White Recessive Rock were generated by deep sequencing, using Illumina Genome Analyzer IIx.

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Unravelling the cellular and humoral immune responses of Antheraea pernyi hemolymph to Antheraea pernyi nucleopolyhedrovirus infection

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