Thoresen Merrilee, Vanover Daryll, King E Heath, Sidelinger Darcie, Feugang Jean M, Peck Hannah E, McAtee Kenzie, Santangelo Philip J, Woolums Amelia R
Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS, USA.
Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
Exp Parasitol. 2025 Aug;275:108985. doi: 10.1016/j.exppara.2025.108985. Epub 2025 Jul 1.
Bovine trichomonosis is caused by the urogenital parasite Tritrichomonas foetus (T. foetus). In the United States, approved therapies are lacking, and management is limited to culling infected bulls. Preputial therapy with synthetic mRNA could lead to effective new treatments. We developed synthetic mRNA encoding bovine IgG1 against two epitopes of the T. foetus cell surface antigen TF1.17 and used the mRNA to transfect bovine cells in vitro. Transfected cells expressed membrane anchored or secreted versions of the antibodies with a NanoLuciferase (NanoLuc) reporter molecule fused to each light chain. Luminescence in cells and supernatants collected 24 and 48 h post-transfection confirmed the production of anti-TF1.17 and was significantly higher than in non-transfected controls (p < 0.05). Anti-TF1.17 bound to live parasites as indicated by significantly higher luminescence following treatment with 24 and 48 h post-transfection supernatants compared to transfection controls (p = 0.001). Treatment of T. foetus with concentrated anti-TF1.17 antibody decreased parasite viability. When T. foetus were added to mRNA transfected kidney cells 48 h post transfection, cytopathic effects of the parasites were reduced following 24 h of co-culture with cells producing anti-TF1.17 as compared to controls (p < 0.05). To our knowledge, this is the first use of mRNA transfection of bovine cells to induce the expression of antibodies that can bind to T. foetus, decrease their viability and their cytopathic effects on host cells. This work forms the basis for the development of novel mRNA-mediated approaches to treat or prevent bovine trichomonosis.
牛毛滴虫病由泌尿生殖系统寄生虫胎儿三毛滴虫（T. foetus）引起。在美国，缺乏经批准的治疗方法，管理措施仅限于淘汰感染公牛。用合成mRNA进行包皮治疗可能会带来有效的新疗法。我们开发了编码针对胎儿三毛滴虫细胞表面抗原TF1.17两个表位的牛IgG1的合成mRNA，并使用该mRNA在体外转染牛细胞。转染细胞表达了与每个轻链融合有纳米荧光素酶（NanoLuc）报告分子的膜锚定或分泌型抗体。转染后24小时和48小时收集的细胞和上清液中的发光证实了抗TF1.17的产生，且显著高于未转染的对照（p < 0.05）。与转染对照相比，用转染后24小时和48小时的上清液处理后，发光显著更高，表明抗TF1.17与活寄生虫结合（p = 0.001）。用浓缩的抗TF1.17抗体处理胎儿三毛滴虫可降低寄生虫活力。在转染后48小时将胎儿三毛滴虫添加到mRNA转染的肾细胞中，与对照相比，与产生抗TF1.17的细胞共培养24小时后，寄生虫的细胞病变效应降低（p < 0.05）。据我们所知，这是首次利用牛细胞的mRNA转染来诱导能够结合胎儿三毛滴虫、降低其活力及其对宿主细胞的细胞病变效应的抗体表达。这项工作为开发治疗或预防牛毛滴虫病的新型mRNA介导方法奠定了基础。
