Expression of MHC class I pathway genes in response to infectious salmon anaemia virus in Atlantic salmon (Salmo salar L.) cells.
Infectious salmon anaemia virus (ISAV) is the causative agent of an important viral disease threatening Atlantic salmon aquaculture. Although its structure and pathogenesis is well described little is known about its immunomodulatory effects on the host. Cellular immunity is critical in the host control of virus infections, an event attributable to antigen presentation through the MHC class I pathway, whose genes are transcriptionally activated by interferons (IFN) and other cytokines. In this study we analysed the regulation and kinetics of key genes in the salmon MHC class I pathway in relation to type I IFN during ISAV infection and poly I:C stimulation in the permissive Atlantic salmon kidney cell line (ASK). As measured by quantitative real-time PCR, ISAV induced an mRNA shut-off equivalent to 2.5-5.5-fold reduced levels of housekeeping genes at 7 days post infection. Relative to this shut-off (by normalising to beta-actin) transcription increased to peak levels at 2.8-fold for MHC class I, 10-fold for beta 2 microglobulin (beta 2m), 5.9-fold for the peptide transporter ABCB2, 8.8-fold for the proteasome component PSMB8 and 4.6-fold for the proteasome component PSMB9, presumably by activation of the IFN system as a 26-fold induction was observed for type I IFN-alpha. Expression of Mx protein was also induced 17-fold at peak level. Similar kinetics and activation levels of these genes were seen in poly I:C stimulated cells. We also isolated the salmon MHC class I UBA*0301 promoter and identified a conserved interferon-stimulated response element (ISRE) and GAAA-elements plus several GAS- and IRF-sites, all supporting IFN-inducible properties. In summary, we demonstrate a concerted induction of the MHC class I pathway and type I IFN by ISAV comparable to levels induced by the synthetic double-stranded RNA (dsRNA) poly I:C. Thus, unlike influenza and several other viruses ISAV does not seem to interfere with MHC and IFN expression.
Jørgensen
Sven Martin
SM
Department of Pharmaceutical Biosciences, School of Pharmacy, University of Oslo, 0316 Oslo, Norway.
Syvertsen
Berit Lyng
BL
Lyng-Syvertsen
Berit
B
Lukacs
Morten
M
Grimholt
Unni
U
Gjøen
Tor
T
D000818:Animals; D001483:Base Sequence / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers / Q000737:chemistry; D005393:Fish Diseases / Q000276:immunology / Q000821:virology; D019204:GTP-Binding Proteins / Q000032:analysis / Q000096:biosynthesis; D015967:Gene Expression Regulation, Viral / Q000276:immunology*; D005805:Genes, MHC Class I / Q000276:immunology*; D016898:Interferon-alpha / Q000032:analysis / Q000096:biosynthesis; D046829:Isavirus / Q000235:genetics / Q000276:immunology*; D007668:Kidney / Q000166:cytology; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000662:veterinary; D011070:Poly I-C / Q000276:immunology; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D020317:Salmo salar / Q000276:immunology* / Q000821:virology; D013997:Time Factors
D017931:DNA Primers; D016898:Interferon-alpha; D064547:Myxovirus Resistance Proteins; D012333:RNA, Messenger; D019204:GTP-Binding Proteins; D011070:Poly I-C