Development and use of an Arctic charr cell line to study antiviral responses at extremely low temperatures

Abstract

Arctic charr (Salvelinus alpinus) are the northernmost distributed freshwater fish and can grow at water temperatures as low as 0.2 °C. Other teleost species have impaired immune function at temperatures that Arctic charr thrive in, and thus, charr may maintain immune function at these temperatures. In this study, a fibroblastic cell line, named ACBA, derived from the bulbus arteriosus (BA) of Arctic charr was developed for use in immune studies at various temperatures. ACBA has undergone more than forty passages at 18 °C over 3 years, while showing no signs of senescence-associated β-galactosidase activity and producing nitric oxide. Remarkably, ACBA cells survived and maintained some mitotic activity even at 1 °C for over 3 months. At these low temperatures, ACBA also continued to produce MH class I proteins. After challenge with poly I:C, only antiviral Mx proteins were induced while MH proteins remained constant. When exposed to live viruses, ACBA was shown to permit viral infection and replication of IPNV, VHSV IVa and CSV at 14 °C. Yet at the preferred temperature of 4 °C, only VHSV IVa was shown to replicate within ACBA. This study provides evidence that Arctic charr cells can maintain immune function while also resisting infection with intracellular pathogens at low temperatures.

Transcriptomic Profiling of the Adaptive and Innate Immune Responses of Atlantic Salmon to Renibacterium salmoninarum Infection

“…As shown in Figure 13, our transcriptome profiling identified several R. salmoninarum-responsive transcripts involved in antigen-presenting and processing pathways. The teleost MH-I-dependent pathway exhibits several differences with their mammalian counterparts (123)(124)(125)(126)(127)(128); for example, different paralogues (i.e., Calnexin) and splice variants (i.e., Tapasin) of molecules involved in the MH-I pathway were identified and suggested to play diverged or unique roles in salmonid antigen presentation (123)(124)(125)(126)(127)(128). The qPCR results showed the R. salmoninarum-dependent up-regulation of fcgr1 and tnfsf14, as well as down-regulation of cdh1, flrt3, and ndrg2 (Figure 9).…”

Antimicrobial Peptides of Salmonid Fish: From Form to Function

“…The research surrounding AMP gene expression during IHNV infection in other salmonids besides rainbow trout and Sockeye salmon remains unknown. Our lab has generated a continuous cell line from Arctic Char, known as ACBA, and has confirmed its susceptibly to VHSV and IHNV replication; however, the AMP gene expression profile of this cell line during infection remains unknown [ 65 ]. Given that various AMPs, including hepcidin, have been detected in Arctic Char at the transcript level, it is possible that ACBA may express AMPs [ 66 ].…”

A More Open Approach Is Needed to Develop Cell-Based Fish Technology: It Starts with Zebrafish

“…Some studies suggest that cultured fish cells exhibit senescence and have proliferation potential similar to mammalian cells. [42][43][44] However, other studies have found that fish cells are less susceptible to senescence and are able to undergo many more doublings [45][46][47][48][49][50][51][52][53] (Figure 2A). With specific regard to muscle stem cells, adult fish satellite cells isolated from trout have been shown to be proliferative in vitro, in contrast to quiescent satellite cells in adult mice.…”