Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 Utrecht CL, The Netherlands.
Virol J. 2013 Jun 24;10:206. doi: 10.1186/1743-422X-10-206.
Influenza viruses are characterized by their highly variable surface proteins HA and NA. The third surface protein M2 is a nearly invariant protein in all Influenza A strains. Despite extensive studies in other animal models, this study is the first to describe the use of recombinant M2 protein and a peptide coding for the extracellular part of the M2 protein (M2e) to vaccinate poultry.
Four groups of layer chickens received a prime-boost vaccination with recombinant M2 protein, M2e, a tetrameric construct from M2e peptide bound to streptavidin and a control tetrameric construct formulated with Stimune adjuvant.
We determined the M2-specific antibody (Ab) responses in the serum before vaccination, three weeks after vaccination and two weeks after booster, at days 21, 42 and 56 of age. The group vaccinated with the M2 protein in combination with Stimune adjuvant showed a significant Ab response to the complete M2 protein as compared to the other groups. In addition an increased Ab response to M2e peptide was found in the group vaccinated with the M2e tetrameric construct. None of the vaccinated animals showed seroconversion to AI in a commercial ELISA. Finally no Ab's were found that bound to M2 expressed on in vitro AI infected MDCK cells.
Although Ab's are formed against the M2 protein and to Streptavidin bound M2e peptide in a tetrameric conformation these Ab's do not recognize of M2 on the virus or on infected cells.
流感病毒的表面蛋白 HA 和 NA 具有高度变异性。第三个表面蛋白 M2 在所有流感 A 株中都是几乎不变的蛋白。尽管在其他动物模型中进行了广泛的研究，但这项研究是首次描述使用重组 M2 蛋白和编码 M2 蛋白胞外部分的肽（M2e）来给家禽接种疫苗。
四组蛋鸡接受了重组 M2 蛋白、M2e、与链霉亲和素结合的 M2e 肽的四聚体构建体和用 Stimune 佐剂配制的对照四聚体构建体的初免-加强免疫接种。
我们在接种前、接种后三周和加强接种后两周，即 21、42 和 56 日龄时，测定了血清中的 M2 特异性抗体（Ab）反应。与其他组相比，用 M2 蛋白与 Stimune 佐剂联合免疫的组显示出对完整 M2 蛋白的显著 Ab 反应。此外，在接种 M2e 四聚体构建体的组中发现对 M2e 肽的 Ab 反应增加。在商业 ELISA 中，没有接种动物对 AI 发生血清转换。最后，没有发现与体外感染的 MDCK 细胞上表达的 M2 结合的 Ab。
尽管针对 M2 蛋白和以四聚体形式结合的链霉亲和素的 M2e 肽形成了 Ab，但这些 Ab 不能识别病毒或感染细胞上的 M2。
