A recombinant CHSE-214 cell line expressing an Mx1 promoter-reporter system responds to both interferon type I and type II from salmonids and represents a versatile tool to study the IFN-system in teleost fish.
A transgenic cell line for the detection of salmon interferons (IFNs) has been established. It is based on a CHSE-214 cell line containing a reporter construct expressing firefly luciferase under the control of the rainbow trout promoter for the IFN-induced Mx1 gene. This cell line, named CHSE-Mx10, showed IFN-induced luciferase expression after more than 80 passages, confirming the stability of this cell line. Interestingly, the Mx promoter was shown to respond to both salmon IFN-alpha/beta and trout IFN-gamma in a dose-dependent manner, while there was no response to TNF-alpha and IL-1beta. IFN-alpha/beta activity could be measured at a range of 9-150 U/ml, and IFN-gamma showed activity between 10 and 100 ng/ml. The reproducibility of both responses was good. The CHSE-Mx10 reporter system constitutes a versatile tool to study the induction and regulation of IFN signaling in teleost fish. A preliminary study presented herein suggests that both infectious pancreas necrosis virus (IPNV) and salmon pancreas disease virus (SPDV) may block activation of the Mx promoter in CHSE-Mx10 stimulated with IFN-alpha/beta.
Jørgensen
Jorunn B
JB
University of Tromsø, Norwegian College of Fishery Science, Department of Marine Biotechnology, N-9037 Tromsø, Norway. jorunn.jorgensen@nfh.uit.no
Johansen
Audny
A
Hegseth
Marit N
MN
Zou
Jun
J
Robertsen
Børre
B
Collet
Bertrand
B
Secombes
Christopher J
CJ
D000818:Animals; D018175:Birnaviridae Infections / Q000276:immunology; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D005393:Fish Diseases / Q000276:immunology / Q000821:virology; D019204:GTP-Binding Proteins / Q000235:genetics*; D020869:Gene Expression Profiling / Q000662:veterinary; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D018061:Infectious pancreatic necrosis virus / Q000276:immunology; D007370:Interferon Type I / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism*; D049409:Luciferases, Firefly / Q000235:genetics; D064547:Myxovirus Resistance Proteins; D017686:Oncorhynchus mykiss / Q000276:immunology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012474:Salmon / Q000276:immunology*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014035:Togaviridae / Q000276:immunology; D014036:Togaviridae Infections / Q000276:immunology