Kuninaga Naotoshi, Asano Makoto, Matsuyama Ryota, Minemoto Takahiro, Mori Takayuki, Suzuki Masatsugu
The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.
Joint Department of Veterinary Medicine, Faculty of Applied Biological Sciences Gifu University, 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.
J Vet Med Sci. 2019 Feb 28;81(2):328-337. doi: 10.1292/jvms.17-0605. Epub 2018 Dec 27.
The small Indian mongoose (Herpestes auropunctatus) was introduced to Japanese islands and has impacted on the island's biodiversity. Population control has been attempted through capturing but its efficiency has rapidly declined. Therefore, new additional control methods are required. Our focus has been on the immunocontraceptive vaccines, which act in an especially species-specific manner. The amino-acid sequence of the mongoose ovum zona pellucida protein 3 (ZP3) was decoded and two types of synthetic peptides (A and B) were produced. In this study, these peptides were administered to mongooses (each n=3) and the sera were collected to verify immunogenicity using ELISA and IHC. Treated mongoose sera showed an increasing of antibody titer according to immunizations and the antigen-antibody reactions against the endogenous mongoose ZP. In addition, IHC revealed that immune sera absorbed with each peptide showed a marked reduction in reactivity, which indicated the specificity of induced antibodies. These reactions were marked in peptide A treated mongoose sera, and the antibody titer of one of them lasted for at least 21 weeks. These results indicated that peptide A was a potential antigen, inducing autoantibody generation. Moreover, immunized rabbit antibodies recognized mongoose ZP species-specifically. However, the induction of robust immune memory was not observed. Also, the actual sterility effects of peptides remain unknown, it should be verified as a next step. In any case, this study verified synthetic peptides we developed are useful as the antigen candidates for immunocontraception of mongooses.
小印度猫鼬(Herpestes auropunctatus)被引入日本岛屿,对该岛的生物多样性造成了影响。人们曾试图通过捕捉来控制其数量,但效率迅速下降。因此,需要新的额外控制方法。我们的重点一直放在免疫避孕疫苗上,这种疫苗具有特别的物种特异性作用方式。猫鼬卵透明带蛋白3(ZP3)的氨基酸序列被解码,并制备了两种合成肽(A和B)。在本研究中,将这些肽注射给猫鼬(每组n = 3),并收集血清,使用酶联免疫吸附测定(ELISA)和免疫组织化学(IHC)来验证免疫原性。经处理的猫鼬血清显示,随着免疫次数的增加,抗体滴度升高,并且与内源性猫鼬ZP发生抗原 - 抗体反应。此外,免疫组织化学显示,用每种肽吸收后的免疫血清反应性显著降低,这表明诱导抗体具有特异性。这些反应在肽A处理的猫鼬血清中很明显,其中一只猫鼬的抗体滴度至少持续了21周。这些结果表明,肽A是一种潜在抗原,可诱导自身抗体产生。此外,免疫兔抗体对猫鼬ZP具有物种特异性识别。然而,未观察到强大免疫记忆的诱导。而且,肽的实际不育效果仍然未知,下一步应进行验证。无论如何,本研究验证了我们开发的合成肽作为猫鼬免疫避孕抗原候选物是有用的。