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表达 J 亚群禽白血病病毒 Gp85 蛋白的重组侵袭性植物乳杆菌诱导鸡对禽白血病的保护。

Recombinant invasive Lactobacillus plantarum expressing the J subgroup avian leukosis virus Gp85 protein induces protection against avian leukosis in chickens.

机构信息

College of Animal Science, Yangtze University, Jingzhou, 434025, China.

College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education, Jilin Agricultural University, Changchun, 130118, China.

出版信息

Appl Microbiol Biotechnol. 2022 Jan;106(2):729-742. doi: 10.1007/s00253-021-11699-9. Epub 2021 Dec 31.

Abstract

Avian leukosis, caused by avian leukosis virus (ALV), is an infectious tumor disease and severely hinders the development of the poultry industry. The use of Lactobacillus plantarum (L. plantarum) could effectively alleviate viremia in the early period of J subgroup ALV (ALV-J) infection. In this study, an invasive L. plantarum NC8 expressing Gp85 protein of ALV-J was constructed. After chickens were orally administered the recombinant invasive NC8, the levels of expression of CD4 and CD8 T lymphocytes in peripheral blood and spleen by flow cytometry and the proliferation ability of splenocytes by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay were examined, and the contents of cytokines, the anti-ALV-J antibody in serum, and mucosal antibody sIgA in intestinal lavage fluid were detected by enzyme-linked immunosorbent assay (ELISA). The immunoprotective efficiency was evaluated by monitoring the infection rate, the percent of cloacal swabs and survival, body weight gain, the organ indexes, and relative virus loads after challenge with ALV-J. The results showed that the recombinant invasive strain (FnBPA-gp85) could promote the expression levels of the CD8T cells in peripheral blood and spleen, the proliferation of splenocytes, the secretions of cytokines interleukin 2 (IL-2) and γ-interferon (IFN-γ), and the production of IgG and sIgA compared with the PBS and FnBPA control groups in chickens. The FnBPA-gp85 group was exhibited the highest immune protection against ALV-J infection. The above results indicated that the recombinant invasive NC8 could promote the cellular immunity, humoral immunity, and mucosal immunity responses in chicken and provide a new method for exploring the live vaccine against ALV-J.Key points• The FnBPA-gp85 strain could enhance cellular immunity response.• The FnBPA-gp85 strain could improve the immune protection against ALV-J infection.

摘要

禽白血病,由禽白血病病毒(ALV)引起,是一种传染性肿瘤病,严重阻碍了家禽业的发展。使用植物乳杆菌(L. plantarum)可以有效缓解 J 亚群 ALV(ALV-J)感染早期的病毒血症。本研究构建了一株能表达 ALV-J 囊膜糖蛋白 Gp85 的侵袭性植物乳杆菌 NC8。经鸡口服重组侵袭性 NC8 后,流式细胞术检测外周血和脾脏中 CD4 和 CD8 T 淋巴细胞的表达水平,3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法检测脾淋巴细胞的增殖能力,酶联免疫吸附试验(ELISA)检测血清中细胞因子、抗 ALV-J 抗体和肠灌洗液黏膜抗体 sIgA 的含量。通过监测攻毒后感染率、泄殖腔拭子阳性率和存活率、体重增长、器官指数和相对病毒载量,评估免疫保护效率。结果表明,与 PBS 和 FnBPA 对照组相比,重组侵袭株(FnBPA-gp85)能促进鸡外周血和脾脏中 CD8T 细胞表达水平、脾淋巴细胞增殖、细胞因子白细胞介素 2(IL-2)和γ-干扰素(IFN-γ)分泌以及 IgG 和 sIgA 的产生。FnBPA-gp85 组对 ALV-J 感染的免疫保护作用最强。上述结果表明,重组侵袭性 NC8 能促进鸡的细胞免疫、体液免疫和黏膜免疫反应,为探索 ALV-J 活疫苗提供了新方法。

关键词

  • FnBPA-gp85 株可增强细胞免疫应答。

  • FnBPA-gp85 株可提高对 ALV-J 感染的免疫保护。

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