Mai Yong-Zhan, Li Yan-Wei, Li Rui-Jun, Li Wei, Huang Xia-Zi, Mo Ze-Quan, Li An-Xing
State Key Laboratory of Biocontrol, Guangdong Province Key Laboratory for Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China.
College of Animal Science, South China Agricultural University, Guangzhou, China.
Vet Parasitol. 2015 Jun 30;211(1-2):1-11. doi: 10.1016/j.vetpar.2015.05.004. Epub 2015 May 14.
Cryptocaryoniasis is a severe disease of farmed marine fish caused by the parasitic ciliate Cryptocaryon irritans. This disease can lead to considerable economic loss, but studies on proteins linked to disease development and antigenic proteins for vaccine development have been relatively scarce to date. In this study, 53 protein spots with differential abundance, representing 12 proteins, were identified based on a pair-wise comparison among theronts, trophonts, and tomonts. Meanwhile, 33 protein spots that elicited serological responses in rabbits were identified, representing 9 proteins. In addition, 27 common antigenic protein spots reacted with grouper anti-sera, representing 10 proteins. Most of the identified proteins were involved in cytoskeletal and metabolic pathways. Among these proteins, actin and α-tubulin appeared in all three developmental stages with differences in molecular weights and isoelectric points; 4 proteins (vacuolar ATP synthase catalytic subunit α, mcm2-3-5 family protein, 26S proteasome subunit P45 family protein and dnaK protein) were highly expressed only in theronts; while protein kinase domain containing protein and heat shock protein 70 showed high levels of expression only in trophonts and tomonts, respectively. Moreover, actin was co-detected with 3 rabbit anti-sera while β-tubulin, V-type ATPase α subunit family protein, heat shock protein 70, mitochondrial-type hsp70, and dnaK proteins showed immunoreactivity with corresponding rabbit anti-sera in theronts, trophonts, and tomonts. Furthermore, β-tubulin, the metabolic-related protein enolase, NADH-ubiquinone oxidoreductase 75 kDa subunit, malate dehydrogenase, as well as polypyrimidine tract-binding protein, glutamine synthetase, protein kinase domain containing protein, TNFR/NGFR cysteine-rich region family protein, and vacuolar ATP synthase catalytic subunit α, were commonly detected by grouper anti-sera. Therefore, these findings could contribute to an understanding of the differences in gene expression and phenotypes among the different stages of parasitic infection, and might be considered as a source of candidate proteins for disease diagnosis and vaccine development.
刺激隐核虫病是由寄生性纤毛虫刺激隐核虫引起的养殖海水鱼类的一种严重疾病。这种疾病会导致相当大的经济损失，但迄今为止，关于与疾病发展相关的蛋白质以及用于疫苗开发的抗原蛋白的研究相对较少。在本研究中，基于对幼虫、滋养体和包囊体的两两比较，鉴定出53个丰度有差异的蛋白点，代表12种蛋白质。同时，鉴定出33个在兔体内引发血清学反应的蛋白点，代表9种蛋白质。此外，27个常见的抗原蛋白点与石斑鱼抗血清发生反应，代表10种蛋白质。大多数鉴定出的蛋白质参与细胞骨架和代谢途径。在这些蛋白质中，肌动蛋白和α-微管蛋白出现在所有三个发育阶段，分子量和等电点存在差异；4种蛋白质（液泡ATP合酶催化亚基α、Mcm2 - 3 - 5家族蛋白、26S蛋白酶体亚基P45家族蛋白和dnaK蛋白）仅在幼虫中高表达；而含蛋白激酶结构域的蛋白和热休克蛋白70分别仅在滋养体和包囊体中高表达。此外，肌动蛋白与3种兔抗血清共同检测到，而β-微管蛋白、V型ATP酶α亚基家族蛋白、热休克蛋白70、线粒体型hsp70和dnaK蛋白在幼虫、滋养体和包囊体中与相应的兔抗血清表现出免疫反应性。此外，β-微管蛋白、与代谢相关的烯醇化酶、NADH - 泛醌氧化还原酶75 kDa亚基、苹果酸脱氢酶，以及多聚嘧啶序列结合蛋白、谷氨酰胺合成酶、含蛋白激酶结构域的蛋白、TNFR/NGFR富含半胱氨酸区域家族蛋白和液泡ATP合酶催化亚基α，均被石斑鱼抗血清共同检测到。因此，这些发现有助于了解寄生虫感染不同阶段的基因表达和表型差异，并可能被视为疾病诊断和疫苗开发的候选蛋白来源。
