Nualart Daniela P, Dann Francisco, Oyarzún-Salazar Ricardo, Morera Francisco J, Vargas-Chacoff Luis
Fish Physiology Laboratory, Institute of Marine and Limnological Sciences, Faculty of Sciences, Universidad Austral de Chile, Valdivia 5090000, Chile.
Ph.D. Program in Aquaculture Sciences, Universidad Austral de Chile, Puerto Montt 5480000, Chile.
Biology (Basel). 2023 Jun 28;12(7):924. doi: 10.3390/biology12070924.
Fish cell culture is a common in vitro tool for studies in different fields such as virology, toxicology, pathology and immunology of fish. Fish cell cultures are a promising help to study how to diagnose and control relevant viral and intracellular bacterial infections in aquaculture. They can also be used for developing vaccines and immunostimulants, especially with the ethical demand aiming to reduce and replace the number of fish used in research. This study aimed to isolate head kidney primary cell cultures from three Chilean salmonids: , , and , and characterize the response to bacterial and viral stimuli by evaluating various markers of the innate and adaptive immune response. Specifically, the primary cell cultures of the head kidney from the three salmonids studied were cultured and exposed to two substances that mimic molecular patterns of different pathogens, i.e., Lipopolysaccharide (LPS) (bacterial) and Polyinosinic: polycytidylic acid (POLY I:C). Subsequently, we determined the mRNA expression profiles of the TLR-1, TLR-8, IgM, TLR-5, and MHC II genes. Head kidney primary cell cultures from the three species grown in vitro responded differently to POLY I:C and LPS. This is the first study to demonstrate and characterize the expression of immune genes in head kidney primary cell culture isolated from three salmonid species. It also indicates their potential role in developing immune responses as defense response agents and targets of immunoregulatory factors.
鱼类细胞培养是一种常见的体外工具,用于鱼类病毒学、毒理学、病理学和免疫学等不同领域的研究。鱼类细胞培养有助于研究如何诊断和控制水产养殖中相关的病毒和细胞内细菌感染。它们还可用于开发疫苗和免疫刺激剂,特别是在旨在减少和替代研究中所用鱼类数量的伦理要求下。本研究旨在从三种智利鲑科鱼类( 、 和 )中分离头肾原代细胞培养物,并通过评估先天免疫和适应性免疫反应的各种标志物来表征对细菌和病毒刺激的反应。具体而言,对所研究的三种鲑科鱼类的头肾原代细胞培养物进行培养,并使其暴露于两种模拟不同病原体分子模式的物质,即脂多糖(LPS)(细菌)和聚肌苷酸:聚胞苷酸(POLY I:C)。随后,我们测定了TLR-1、TLR-8、IgM、TLR-5和MHC II基因的mRNA表达谱。在体外培养的这三种物种的头肾原代细胞培养物对POLY I:C和LPS的反应不同。这是第一项证明并表征从三种鲑科鱼类分离的头肾原代细胞培养物中免疫基因表达的研究。它还表明了它们在作为防御反应剂和免疫调节因子靶点的免疫反应发展中的潜在作用。