Chaohui Dai, Li Sun, Weiyun Qin, Ying Liu, Guoqiang Zhu, Shenglong Wu and Wenbin Bao
Yangzhou University, China
Scientific Tracks Abstracts: Biol syst Open Access
Three full-sib individuals of the resistance and susceptibility to E. coli F18 in Meishan piglets were obtained by challenging with the pathogens through feeding F18ab and F18ac strains and then the transcriptome sequencing of intestinal tissue in sensitive and resistant pigs to E. coli F18 was analyzed. The pig small intestinal epithelial cell line (IPEC-J2) with CD14 gene silencing were established by Lentivirus-mediated RNAi and the transcription and protein expression level of TLR4 pathwayrelated genes (MyD88, IFN-α, IL-1β, TLR4 and TNF-α) were detected; the levels of proinflammatory cytokines IL-6, IL-8, IL- 12, MIP-1α and MIP-1β in cell culture supernatants were measured; the E. coli F18ab and F18acâ??s adhesion ability to IPEC-J2 were detected. The transcriptome sequencing results revealed that toll-like receptor 4 (TLR4) signaling pathway especially CD14 gene played important roles in immune process of E. coli F18-invasion. After CD14 gene silencing, the transcription and protein expression levels of IFN-α, IL-1β, TLR4 and TNF-α were significantly down-regulation (P<0.05); the levels of IL-6 and IL-12 in cell supernatants were significantly reduced (P<0.05); the E. coli F18abâ??s adhesion to IPEC-J2 enhanced highly significantly (P<0.01). In summary, the establishment of IPEC-J2 cell line with CD14 gene silencing stably mediated by Lentivirus offered important material for mechanism research of CD14 gene and TLR4 signal pathway. These results suggested that CD14 gene played an important role in not only TLR4 signaling pathway but also regulating the immune process of E. coli F18-invasion.
Chaohui Dai is currently a Postgraduate student of Yangzhou University, China. She is mainly engaged in pig disease-resistant breeding and reproduction.
Email: chdai1993@163.com
