禽致病性大肠杆菌Hcp2b对雏鸡气管黏膜细胞因子-细胞因子受体相互作用通路的影响

doi:10.11843/j.issn.0366-6964.2021.03.018

Abstract

Abstract: T6SS (type VI secretion system) is a common secretion system in Gram-negative bacteria. And its mechanism of effector protein Hcp2b is still unclear. The Hcp2b protein of avian pathogenic Escherichia coli (APEC) is the main subject of this study, which aims to explore the role and mechanism of Hcp2b protein in the process of APEC infection of chicken tracheal mucosa. The Red recombination method was used to construct the hcp2b-deleted strain by using the plasmid pKD3 as the template. And the pSTV-28 plasmid was connected to the hcp2b target fragment to construct the recombinant vector, which was introduced to the competent Δhcp2b strain to construct the hcp2b-reverted strain. Then, we measured the growth curve of Δhcp2b strain. Seven-day-old chicks were infected with the hcp2b-deleted strain and the wild strain by tracheal injection. The infected tracheal mucosal cells were collected at 12 and 24 h after infection and subjected to transcriptomics sequencing and bioinformatics analysis. The results showed that the Δhcp2b strain and Chcp2b strain were successfully constructed. And the deletion of hcp2b gene did not affect the growth performance of the strain. The expression profile of the mRNA of the tracheal mucosa changed after the hcp2b gene-deleted strain infected. Twelve hours after infection, the expression of 144 genes changed (87 differential genes were up-regulated and 57 were down-regulated); Twenty-four hours after infection, 135 gene expressions changed (79 differential genes up-regulated and 56 down-regulated). Bioinformatics analysis found that differential genes were enriched in signaling pathways such as Cytokine-cytokine receptor interaction, Protein processing in endoplasmic reticulum, etc. The deletion of hcp2b gene does not affect the growth characteristics of APEC. After the deletion of hcp2b gene, it affects the change of mRNA transcription level of Cytokine-cytokine receptor interaction pathway gene in chick tracheal mucosa. And the expression of IL-1β and IL-12b are all up-regulated. This study provides a theoretical basis for the study of the pathogenic mechanism of APEC.

Key words: avian pathogenic Escherichia coli, APEC, hcp2b, tracheal mucosal cells, transcriptome

CLC Number:

S852.612

SONG Xiangjun, SHEN Xiao, JIANG Huyan, CHEN Zhe, LIU Hua, SHAO Ying, TU Jian, QI Kezong. Effect of Avian Pathogenic Escherichia coli Hcp2b on the Cytokine-Cytokine Receptor Interaction Pathway in Chick Tracheal Mucosa[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 742-751.

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Effect of Avian Pathogenic Escherichia coli Hcp2b on the Cytokine-Cytokine Receptor Interaction Pathway in Chick Tracheal Mucosa

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