[含禽流感病毒N端M2e的传染性法氏囊病病毒重组VP2免疫原性的鉴定]

[Characterization the immunogenicity of recombinant VP2 of infectious bursal disease virus containing N-terminal M2e of avian influenza virus].

作者信息

Tang Yinghua, Gong Yuzhen, Wang Yongwei, Lu Jihu, Wu Peipei, Gao Feng, Chen Tao, He Jiahui, Hou Jibo

机构信息

Jiangsu Academy of Agricultural Sciences, National Research Center of Engineering and Technology for Veterinary Bioproducts, Nanjing 210014, China.

出版信息

Wei Sheng Wu Xue Bao. 2012 Jun 4;52(6):753-9.

PMID:22934356

Abstract

OBJECTIVE

We developed subunit vaccines against H5 or H9 subtype avian influenza viruses (AIV) and infectious bursal disease viruses (IBDV). Viral protein 2 (VP2) of IBDV was used as cargo protein to display a 12-amino-acid (aa) immunodominant epitope derived from N-terminal M2 extracelluar domain (nM2e) of H5 or H9 subtype AIV.

METHODS

The aa and nucleotide sequence of nM2e was determined by comparing the available avian influenza vaccine strains and alignment the AIV sequence available in GenBank. One copy of H5 or H9 nM2e was inserted into P(BC) region of VP2 origin from IBDV B87 vaccine strain by fusion polymerase chain reaction. The VP2(BC)nM2e recombinants were cloned into Bac-to-Bac expression system and transfected to Sf9 cell. The expressed chimeric protein was characterized by indirect immunofluorescence assay and Western blotting, and subsequently was used as antigen to develop vaccine. The non-immunized chicken was given two injections with the vaccine at a 4-week interval. Serum against VP2 and nM2e was tested by indirect ELISA and virus neutralization in chick embryo fibroblast.

RESULTS

Both VP2(BC)nM2e recombinants were successfully constructed and expressed in Sf9 cell. Both chimeric proteins elicited antibody against VP2 and nM2e. The antibody level elicited by VP2(BC)nM2e(H5) vaccine was higher than that of VP2(BC)nM2e(H9).

CONCLUSION

Both chimeric proteins were immunigenic, and the efficacy of VP2(BC)nM2e(H5) was higher than VP2(BC)nM2e(H9) chicken.

摘要

目的

我们研发了针对H5或H9亚型禽流感病毒（AIV）以及传染性法氏囊病病毒（IBDV）的亚单位疫苗。IBDV的病毒蛋白2（VP2）被用作载体蛋白，以展示源自H5或H9亚型AIV的N端M2细胞外结构域（nM2e）的一个12氨基酸（aa）的免疫显性表位。

方法

通过比较现有的禽流感疫苗株并比对GenBank中可用的AIV序列，确定nM2e的氨基酸和核苷酸序列。通过融合聚合酶链反应，将一份H5或H9 nM2e插入源自IBDV B87疫苗株的VP2的P(BC)区域。将VP2(BC)nM2e重组体克隆到杆状病毒表达系统中，并转染至Sf9细胞。通过间接免疫荧光测定和蛋白质印迹对表达的嵌合蛋白进行表征，随后将其用作抗原研发疫苗。未免疫的鸡每隔4周接种两次该疫苗。通过间接ELISA和鸡胚成纤维细胞中的病毒中和试验检测针对VP2和nM2e的血清。