A comparison of rainbow trout cell lines for their expression of the major histocompatibility complex genes and the induction of beta-2-microglobulin by dsRNA

Kawano, Atsushi; Kales, Stephen C.; Fujiki, Kazuhiro; DeWitte‐Orr, Stephanie J.; Dixon, Brian; Lee, Lucy E. J.; Bols, Niels C.

doi:10.1016/j.fsi.2010.04.007

Cited by 12 publications

(14 citation statements)

References 53 publications

(57 reference statements)

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“…Currently, several immortalized cells derived from various epithelial lining tissues are available for studying immune gene expression in trout (Bols et al, 1994;Kawano et al, 2011). Recently, the RTgutGC cell line from the intestinal epithelia of Oncorhynchus mykiss was shown to express several immune related molecules, such as TNF-α (Kawano et al, 2011), tapasin (Sever et al, 2014), MHC class I alpha chain and beta-2-microglobulin (Kawano et al, 2010), suggesting behavior comparable to IECs. Therefore, RTgutGC cells were considered a representative model that may be used to address the in vitro effect of zymosan in the intestinal immune response of the rainbow trout.…”

Section: Discussionmentioning

confidence: 99%

Immunomodulatory effect of cathelicidins in response to a β-glucan in intestinal epithelial cells from rainbow trout

Schmitt

Wacyk

Morales‐Lange

et al. 2015

Developmental & Comparative Immunology

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Section: Discussionmentioning

confidence: 99%

Immunomodulatory effect of cathelicidins in response to a β-glucan in intestinal epithelial cells from rainbow trout

Schmitt

Wacyk

Morales‐Lange

et al. 2015

Developmental & Comparative Immunology

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“…Previously, this anti‐trout MH Iα antibody was shown to cross‐react with a Chinook salmon cell line (CHSE‐214) (Kawano et al . ), but this is the first demonstration for cross‐reaction to be demonstrated with a salmonid species in the genus Salvelinus .…”

Section: Resultsmentioning

confidence: 81%

“…To study whether ACBA cells produce MH class I proteins, affinity‐purified rabbit polyclonal antibodies raised against MH class I heavy chain (MH Iα) (Kales ; Kawano et al . ) and beta‐2 microglobulin (β2 m) (Kales et al . ; Kawano et al .…”

Section: Methodsmentioning

confidence: 99%

“…; Kawano et al . ) homologues from another salmonid, rainbow trout, were used. ACBA cells were equally seeded into 75‐cm 2 flasks and were allowed to grow to confluence at 14 °C.…”

Section: Methodsmentioning

confidence: 99%

“…For rainbow trout cell lines examined to date, expression is seen of antiviral proteins, such as Mx (DeWitte‐Orr et al ., ), and components of the endogenous pathway of antigen presentation, such as the major histocompatibility complex I alpha chain (MH Iα) (Kawano et al . ) and ERp57 (Sever, Bols & Dixon ). In response to the viral mimic, poly inosinic: poly cytidylic (poly I:C), some of these are induced whereas others are constitutively expressed (DeWitte‐Orr et al .…”

Section: Introductionmentioning

confidence: 99%

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

Semple

et al. 2017

Journal of Fish Diseases

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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.

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Application of the rainbow trout derived intestinal cell line (RTgutGC) for ecotoxicological studies: molecular and cellular responses following exposure to copper

et al. 2017

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There is an acknowledged need for in vitro fish intestinal model to help understand dietary exposure to chemicals in the aquatic environment. The presence and use of such models is however largely restrictive due to technical difficulties in the culturing of enterocytes in general and the availability of appropriate established cell lines in particular. In this study, the rainbow trout (Oncorhynchus mykiss) intestinal derived cell line (RTgutGC) was used as a surrogate for the “gut sac” method. To facilitate comparison, RTgutGC cells were grown as monolayers (double-seeded) on permeable Transwell supports leading to a two-compartment intestinal model consisting of polarised epithelium. This two-compartment model divides the system into an upper apical (lumen) and a lower basolateral (portal blood) compartment. In our studies, these cells stained weakly for mucosubstances, expressed the tight junction protein ZO-1 in addition to E-cadherin and revealed the presence of polarised epithelium in addition to microvilli protrusions. The cells also revealed a comparable transepithelial electrical resistance (TEER) to the in vivo situation. Importantly, the cell line tolerated apical saline (1:1 ratio) thus mimicking the intact organ to allow assessment of uptake of compounds across the intestine. Following an exposure over 72 h, our study demonstrated that the RTgutGC cell line under sub-lethal concentrations of copper sulphate (Cu) and modified saline solutions demonstrated uptake of the metal with saturation levels comparable to short term ex situ gut sac preparations. Gene expression analysis revealed no significant influence of pH or time on mRNA expression levels of key stress related genes (i.e. CYP3A, GST, mtA, Pgp and SOD) in the Transwell model. However, significant positive correlations were found between all genes investigated suggesting a co-operative relationship amongst the genes studied. When the outlined characteristics of the cell line are combined with the division of compartments, the RTgutGC double seeded model represents a potential animal replacement model for ecotoxicological studies. Overall, this model could be used to study the effects and predict aquatic gastrointestinal permeability of metals and other environmentally relevant contaminants in a cost effective and high throughput manner.

show abstract

A comparison of rainbow trout cell lines for their expression of the major histocompatibility complex genes and the induction of beta-2-microglobulin by dsRNA

Cited by 12 publications

References 53 publications

Immunomodulatory effect of cathelicidins in response to a β-glucan in intestinal epithelial cells from rainbow trout

Immunomodulatory effect of cathelicidins in response to a β-glucan in intestinal epithelial cells from rainbow trout

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

Application of the rainbow trout derived intestinal cell line (RTgutGC) for ecotoxicological studies: molecular and cellular responses following exposure to copper

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