传染性支气管炎病毒对鸡免疫反应及重组禽β-防御素11和12抗病毒作用的研究

doi:10.16431/j.cnki.1671-7236.2023.01.025

Abstract

Abstract: 【Objective】 This experiment was conducted to study the host immune response to respiratory Infectious bronchitis virus virulent strain infection in layer and discover the antiviral effect of recombinant avian β-defensin (AvBDs).【Method】 70 7-day old White Leghorn chickens were randomly devided into two groups, thirty-five chickens in each group respectively.The challenge group was inoculated with respiratory IBV virulent strain (CK/CH/LSD/05I) by a combined intraocular and intranasal route, while the control group was inoculated with PBS by the same way.The clinical symptoms were regularly observed for 14 d and recorded.Five chickens from each group were randomly selected to kill by blood sampling at 12 h, 36 h, 3 d, 7 d and 14 d after the infection.Blood samples were collected to detect the level of serum antibody.Part of trachea and kidney were collected for histopathological examination.RNA was extracted from bursa of Fabricius, spleen, kidney and trachea, which was used to detect the viral load of IBV, the expression of Toll-like receptor (TLRs) and signal transduction molecules, AvBDs by Real-time quantitative PCR.Recombinant AvBD11 and AvBD12 were transfected into chicken embryo kidney (CEK) cells.The supernatant and cells were collected at 6, 12, 24, 36 and 48 h after the infection.RNA was extracted for detecting the viral load by Real-time quantitative PCR.【Result】 After being infected with respiratory IBV virulent strain, chickens showed slight respiratory symptoms, and there was no obvious pathological change in autopsy.The antibody level of the infection group turned positive at 14 d post infection, and the positive rate of respiratory IBV virulent strain group was 100%.The results of Real-time quantitative PCR detection showed that no virus was detected in control group, IBV antigen was only detected in the trachea at 14 d post infection in challenge group and the virus load was very low.Compared with control group, the expression of TLR1, TLR5, AvBD9 and AvBD12 genes in spleen were significantly up-regulated at 7 d post infection with respiratory IBV virulent strain (P<0.05).In challenge group, the expression of TLR1, TLR2, TLR3, AvBD5 and AvBD9 genes in bursa of Fabricius and the expression of nuclear factor-κB p65 (NF-κB p65), NF-κB c-Rel, AvBD5, AvBD9, AvBD11 and AvBD13 genes in trachea were significantly increased at 14 d after the infection (P<0.05).The results of recombinant defensin antiviral experiment showed that compared with the empty vector group, the viral load of CEK cells transfected with AvBD11 was significantly down-regulated at 24 and 48 h after the infection (P<0.05), and the viral load in the supernatant of CEK cells transfected with AvBD11 was significantly down-regulated at 48 h post infection (P<0.05).Compared with the transfected empty vector group, the viral load of CEK cells transfected with AvBD12 was significantly down-regulated at 12 and 24 h after the infection (P<0.05), and the viral load in the supernatant of CEK cells transfected with AvBD12 was significantly down-regulated at 12 and 48 h post infection (P<0.05).It showed that recombinant AvBD11 and AvBD12 had anti-IBV activity in CEK cells.【Conclusion】 The infection of the respiratory IBV virulent strain enhanced the gene expression of relative receptor and signal transduction molecules, upregulated the gene expression of defensins, and strengthened the body's natural immune response.The recombinant defensins AvBD11 and AvBD12 had antiviral effects, which provided a new idea for the preparation and research of non-antiviral feed.

Key words: Infectious bronchitis virus (IBV); chicken; Toll-like receptor; avian β-defensin; antiviral effect

CLC Number:

S852.4

XIE Jingjing, REN Mengting, FAN Jiahui, JIAO Yaru, HAN Zongxi, MA Deying. Study on Immune Response of Infectious Bronchitis Virus to Chickens and Antiviral Effect of Recombinant Avian Beta-defensins 11 and 12[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 246-259.

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Study on Immune Response of Infectious Bronchitis Virus to Chickens and Antiviral Effect of Recombinant Avian Beta-defensins 11 and 12

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