Screening of differentially expressed immune-related genes from spleen of broilers fed with probiotic Bacillus cereus PAS38 based on suppression subtractive hybridization.

Jiajun Li,Dan Xiao,Yufei Wang,Jianzhen Li,Xueqin Ni,Lei Liu,Dong Zeng,Bo Jing,Dongmei Zhang,Zhenhua Wang,Qihui Luo,Wanqiang Li,Kangcheng Pan,Juan J Loor

doi:10.1371/journal.pone.0226829

Abstract

The aim of this study was to construct the spleen differential genes library of broilers fed with probiotic Bacillus cereus PAS38 by suppression subtractive hybridization (SSH) and screen the immune-related genes. Sixty seven-day-old broilers were randomly divided into two groups. The control group was fed with basal diet, and the treated group was fed with basal diet containing Bacillus cereus PAS38 1×106 CFU/g. Spleen tissues were taken and extracted its total RNA at 42 days old, then SSH was used to construct differential gene library and screen immune-related genes. A total of 119 differentially expressed sequence tags (ESTs) were isolated by SSH and 9 immune-related genes were screened out by Gene ontology analysis. Nine differentially expressed genes were identified by qRT-PCR. JCHAIN, FTH1, P2RX7, TLR7, IGF1R, SMAD7, and SLC7A6 were found to be significantly up-regulated in the treated group. Which was consistent with the results of SSH. These findings imply that probiotic Bacillus cereus PAS38-induced differentially expressed genes in spleen might play an important role in the improvement of immunity for broilers, which provided useful information for further understanding of the molecular mechanism of probiotics responsible to affect the poultry immunity.

Highlights

Bacillus spp. is a common probiotic and widely used in poultry industry [1]
Gao et al [15] studied the immune mechanism of silkworm against Bombyxmori nuclear polyhedrosis virus (BmNPV), the differentially expressed gene libraries of resistant strains and susceptible strains were constructed by suppression subtractive hybridization (SSH) technique, and they found that 17 genes were up-regulated in resistant strain BC10, which involved in cell metabolism, transmembrane transport, cytoskeleton, protease, development and immunity, and these genes may be related to resistance to BmNPV in silkworm
In order to study the effect of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) infection on the response of pig cells, SSH was used to compare cDNA libraries of porcine alveolar macrophages (PAM) infected with HP-PRRSV and uninfected HP-PRRSV, and results showed that 21 genes including IL-16, TGF-β type 1 receptor, epidermal growth factor, MHC-I SLA, toll-like receptor, hepatoma-derived growth factor, ferritin heavy chain 1 (FTH1), and MHC-II SLA-DRB1 were up-regulated in PAM infected with HP-PRRSV, by the way, this research demonstrated differential gene expression between HP-PRRSV-infected and uninfected PAMs in vivo for the first time [16]

Summary

Introduction

Bacillus spp. is a common probiotic and widely used in poultry industry [1]. A large number of studies have shown that Bacillus could promote the development of immune organs and activate immune-related signaling pathways serving as an immune activator, thereby stimulating the specific and non-specific immunity of the host and improving the immune ability of animals [2,3,4]. Screening of immune-related genes of broilers fed with Bacillus cereus by SSH (GenBank accession numbers JZ980559JZ980677.)

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