Johnson Wendell C, Hussein Hala E, Capelli-Peixoto Janaina, Laughery Jacob M, Taus Naomi S, Suarez Carlos E, Ueti Massaro W
Animal Diseases Research Unit, USDA-ARS, Pullman, WA 99164, USA.
Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164, USA.
Pathogens. 2022 May 27;11(6):623. doi: 10.3390/pathogens11060623.
Bovine babesiosis is caused by apicomplexan pathogens of the genus , including . This protozoan parasite has a complex life cycle involving dynamic changes to its transcriptome during the transition between the invertebrate and vertebrate hosts. Studying the role of genes upregulated by tick stage parasites has been hindered by the lack of appropriate tools to study parasite gene products in the invertebrate host. Herein, we present tfBbo5480, a transfected cell line, constitutively expressing enhanced green fluorescent protein (eGFP) created by a whole gene replacement transfection strategy, that was capable of completing the parasite's entire life cycle in both the vertebrate and invertebrate hosts. tfBbo5480 was demonstrated to respond to in vitro sexual stage induction and upon acquisition by the female tick vector, , the tick specific kinete stage of tfBbo5480 was detected in tick hemolymph. Larvae from tfBbo5480 exposed female ticks successfully transmitted the transfected parasite to a naïve calf. The development of the whole gene replacement strategy will permit a deeper understanding of the biology of parasite-host-vector triad interactions and facilitate the evaluation of upregulated genes during the parasite's journey through the tick vector leading to new intervention strategies for the control of bovine babesiosis.
牛巴贝斯虫病由巴贝斯虫属的顶复门病原体引起，包括[具体种类未给出]。这种原生动物寄生虫具有复杂的生命周期，在无脊椎动物宿主和脊椎动物宿主之间转换时，其转录组会发生动态变化。由于缺乏在无脊椎动物宿主中研究寄生虫基因产物的合适工具，研究蜱阶段寄生虫上调基因的作用受到了阻碍。在此，我们展示了tfBbo5480，这是一种通过全基因替换转染策略构建的、组成性表达增强型绿色荧光蛋白（eGFP）的转染细胞系，它能够在脊椎动物和无脊椎动物宿主中完成寄生虫的整个生命周期。tfBbo5480被证明对体外性阶段诱导有反应，并且在被雌性蜱载体[蜱的具体种类未给出]获取后，在蜱的血淋巴中检测到了tfBbo5480的蜱特异性动合子阶段。来自暴露于tfBbo5480的雌性蜱的幼虫成功地将转染的寄生虫传播给了未感染的小牛。全基因替换策略的发展将有助于更深入地了解寄生虫 - 宿主 - 载体三元组相互作用的生物学特性，并有助于评估寄生虫在蜱载体中传播过程中上调基因的作用，从而为控制牛巴贝斯虫病带来新的干预策略。
