Partial purification and characterization of RTG-2 fish cell interferon

Abstract

Interferon produced by rainbow trout gonadal cells (RTG-2) was partially purified. The physical, chemical, and biological properties of this in vitro produced fish cell interferon were studied. Purification was achieved by ultracentrifugation, molecular sieve gel chromatography, ion exchange chromatography, and polyacrylamide gel electrophoresis. The isoelectric point of RTG-2 interferon, as determined by CM-Sephadex (C-0O) chromatography, was 7.1. Filtration through Sephadex G-150 showed that RTG-2 interferon had a molecular weight of 94,000. The partially purified material was not sedimented at 105,000 x g for 2 h at 4 C. The fish cell interferon was non-dialyzable and exhibited heat and pH stability. The partially purified material was inactivated by treatment with trypsin or 2-mercaptoethanol, but was resistant to treatment with deoxyribonuclease or ribonuclease. RTG-2 interferon which was induced by infectious pancreatic necrosis virus exhibited antiviral activity against challenge with infectious hematopoietic necrosis virus or infectious pancreatic necrosis virus. Partially purified RTG-2 interferon exhibited greater species specificity than the crude material.

Molecular Size, pH, Temperature Stability, and Ontogeny of Inhibitor(s) of Infectious Pancreatic Necrosis Virus (IPNV) in Normal Rainbow Trout Serum

“…We considered the possibility of the inhibitor's being interferon because some investigators have reported that the size of teleost interferon was between 40 and 94 kDa (De Sena and Rio 1975;Dorson et al 1975;Tamai et al 1993); more recent studies, however, have reported that fish interferon size is between 18 and 25 kDa (Altmann et al 2003;Robertsen et al 2003;Long et al 2004), close to the size range for SI in our studies. Both interferon ␣ and ␤ are known to be stable at 56ЊC and pH 2 (Sano and Nagakura 1982).…”

Substances involved in the natural resistance of fish to infection–A review

“…In addition, the molecular weight of fish interferon is higher than that determined for reptiles and higher vertebrates and as one progresses up the phylogentic scale, a reduction in the molecular weight of interferon occurs. With regard to charge and molecular weight heterogeneity, De Sena & Rio (1975) remarked that fish may produce more than one type of interferon, dependant upon the type of inducer and type of cell stimulated. A comparison of the weights obtained by Dorson, Barde & De Kinkelin (1 975) and De Sena & Rio (1975) for rainbow trout interferon suggests that fish interferon can possibly exist as a polymeric form of the basic subunit.…”

Do fish lymphocytes secrete interferon‐γ?

“…It is well established that fish cells can secrete IFN molecules in response to virus infection of fibroblast or epithelial cell lines in vitro (Gravel! & Malsberger, 1965;DeSena & Rio, 1975;Okamoto et ul., 1983) or in vivo (de Kinkelin et ul., 1982). Protection by these IFNs is non-specific in that it can provide protection against an unrelated virus to that which induced it (de Kinkelin et al, 1982).…”

“…Protection by these IFNs is non-specific in that it can provide protection against an unrelated virus to that which induced it (de Kinkelin et al, 1982). Physicochemical analysis of these fish IFNs has shown they possess a molecular weight of between 26 and 94 kDa, have isoelectric focussing points of 54-7.1, and exhibit pH and temperature stability (Dorson rt ul., 1975;DeSena & Rio, 1975). The cell source.…”