State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Lanzhou, China.
Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonose, Yangzhou University, Yangzhou, China.
Front Cell Infect Microbiol. 2021 Feb 26;11:644983. doi: 10.3389/fcimb.2021.644983. eCollection 2021.
Tropical theileriosis is the disease caused by tick-transmitted apicomplexan parasite , which has ability to transform bovine leukocytes, including B cells, macrophage cells, and dendritic cells. The transformed cells are characterized as uncontrolled proliferation and shared some cancer-like phenotypes. The mechanism of the transformation by is still not understood well. In previous reports, the subtelomere-encoded variable secreted proteins (SVSP) of were considered to contribute to phenotypic changes of the host cell, but the role of SVSP of in host-pathogen relationship remains unknown. In the present study, a member of SVSP family, TA05575 of was selected as the target molecule to analyze its expression profiles in different life cycle stages of by qPCR and investigate its subcellular distribution of different passages of transformed cells using confocal experiments. From the results, the transcription level of TA05575 at schizont stage was significantly higher than the other two life stages of , and the protein of TA05575 was mainly distributed in nucleus of infected cells. In addition, the potential proteins of host cells interacting with TA05575 were screened by Yeast-two hybrid system. The results of Co-IP experiment confirmed that TA05575 interacted with RBMX2-like protein that participated in transcription regulation of cells. In addition, a novel BiFC assay and flow cytometry were carried out, and the results further revealed that TA05575-RBMX2-like pair was directly interacted in cell context. Moreover, this interacting pair was found to distribute in intracellular compartments of HEK293T cells by using confocal microscopy. The results of the present study suggest that TA05575 may contribute for cells transformation due its distribution. According to the function of RBMX2, the interaction of TA05575 and RMMX2-like will provide a new information to further understand the mechanisms of cells transformation by .
热带泰勒虫病是由蜱传播的顶复门寄生虫引起的疾病，它能够转化牛的白细胞，包括 B 细胞、巨噬细胞和树突状细胞。转化细胞的特征是不受控制的增殖，并具有一些类似癌症的表型。的转化机制尚不清楚。在以前的报告中，被认为是宿主细胞表型变化的贡献者，但在宿主-病原体关系中 SVSP 的作用仍然未知。在本研究中，选择 中的一个 SVSP 家族成员 TA05575 作为靶分子，通过 qPCR 分析其在不同生命周期阶段的表达谱，并通过共聚焦实验研究不同传代的转化细胞中 TA05575 的亚细胞分布。结果表明，裂殖体阶段 TA05575 的转录水平明显高于 的另外两个生命阶段，TA05575 蛋白主要分布在感染细胞的核内。此外，通过酵母双杂交系统筛选与 TA05575 相互作用的宿主细胞潜在蛋白。Co-IP 实验结果证实 TA05575 与参与细胞转录调控的 RBMX2 样蛋白相互作用。此外，进行了新的 BiFC 测定和流式细胞术，结果进一步表明 TA05575-RBMX2 样对在细胞环境中直接相互作用。此外，通过共聚焦显微镜发现该相互作用对分布在 HEK293T 细胞的细胞内隔室中。本研究结果表明，TA05575 可能因其分布而有助于细胞转化。根据 RBMX2 的功能，TA05575 和 RMMX2 样的相互作用将为进一步了解由引起的细胞转化机制提供新的信息。
