Siddiki Amam Zonaed, Alam Sabreena, Fuad Bin Hossen Farhan, Alim Md Abdul
Department of Pathology and Parasitology, Chittagong Veterinary and Animal Sciences University (CVASU), Chittagong, Bangladesh.
Department of Biochemistry and Biotechnology, University of Science and Technology Chittagong (USTC), Chittagong, Bangladesh.
PLoS One. 2025 Jan 24;20(1):e0312262. doi: 10.1371/journal.pone.0312262. eCollection 2025.
The three rickettsial parasites- Babesia bovis, Theileria annulata and Anaplasma Marginale are responsible for causing Babesiosis, Theileriosis and Anaplasmosis among cattle. These diseases exist due to spreading of infected ticks. A large number of cattle were found to suffer from mixed infections caused by the three parasites at the same time. Due to these reasons cattle have been devoid of milk production with reduced meat availability. Hence, it is a matter of urgency for the immunity of cattle to exhibit resilience against all three rickettsial parasites. It could be possible if trials are carried out after producing a subunit chimeric vaccine against the rickettsial protozoan parasites and introducing it into the bloodstream of the cattle species. In this paper, we have used the process of reverse vaccinology to conduct a study in which we have developed a multi-epitope subunit chimeric vaccine against three protozoan parasites. We constructed three chimeric vaccine sequences from which only one chimeric vaccine construct was found to be an effective and efficient vaccine which is stable with high solubility and negative allergenicity. Following that, we performed molecular docking of the refined chimeric vaccine construct with Rp-105 and TLR-9. It was observed that the chimeric vaccines interacted with the receptors with high binding energy. Immune simulation was also performed to determine the potentiality of the chimeric vaccine for eliciting an immune response. The best-designed chimeric vaccine construct was then reverse transcribed and adapted for the host E. coli K12 strain which was later inserted into the pET28a (+) vector for the cloning and expression of the vaccine. The study could be a good initiative for the development of an effective chimeric vaccine against bovine parasites.
三种立克次体寄生虫——牛巴贝斯虫、环形泰勒虫和边缘无浆体，是导致牛患巴贝斯虫病、泰勒虫病和无浆体病的病原体。这些疾病是由感染蜱虫的传播而存在的。大量牛被发现同时感染了这三种寄生虫。由于这些原因，牛的产奶量下降，可获得的肉类减少。因此，提高牛对所有三种立克次体寄生虫的免疫力迫在眉睫。如果在生产针对立克次体原生动物寄生虫的亚单位嵌合疫苗并将其引入牛的血液中后进行试验，这是有可能实现的。在本文中，我们利用反向疫苗学方法进行了一项研究，开发了一种针对三种原生动物寄生虫的多表位亚单位嵌合疫苗。我们构建了三种嵌合疫苗序列，其中只有一种嵌合疫苗构建体被发现是一种有效且高效的疫苗，具有高稳定性、高溶解性和低致敏性。随后，我们对优化后的嵌合疫苗构建体与Rp - 105和TLR - 9进行了分子对接。观察到嵌合疫苗与受体以高结合能相互作用。还进行了免疫模拟，以确定嵌合疫苗引发免疫反应的潜力。然后将设计最佳的嵌合疫苗构建体进行反转录，并适配宿主大肠杆菌K12菌株，随后将其插入pET28a(+)载体中进行疫苗的克隆和表达。这项研究对于开发一种有效的抗牛寄生虫嵌合疫苗可能是一个很好的开端。
