Cloning and biologic activities of a bovine interferon-alpha isolated from the epithelium of a rotavirus-infected calf.
A cDNA encoding a distinct bovine (Bo) interferon (IFN) alpha, designated BoIFN-alpha E, was generated from gut epithelial cells isolated from a rotavirus-infected calf. The BoIFN-alpha E cDNA sequence shared a greater than 90% identity with the other BoIFN-alpha subtypes. The cDNA encoding BoIFN-alpha E has been expressed in insect cells using the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) as a vector. Insect cells infected with recombinant virus secreted a protein with a relative molecular mass of 19,500 into the culture medium not observed in cells infected with wild-type AcMNPV. Supernatants harvested from cultures of insect cells infected with the recombinant AcMNPV encoding IFN-alpha E inhibited the replication of Semliki Forest virus in a bovine cell line and typically showed 10(6) dilution units/ml of antiviral activity. However, differences were observed between the activities of recombinant BoIFN-alpha E and BoIFN-alpha 1 1 on the proliferation of WC1+ gamma/delta T cells. Purified ( > 99%) WC1+ gamma/delta T cells failed to proliferate to IFN-alpha 1 1 or concanavalin A and IFN-alpha E acted as a weak proliferative signal to these cells, demonstrating a functional difference between two closely related BoIFN-alpha subtypes.
Chaplin
P J
PJ
Division of Immunology and Pathology, Institute for Animal Health, Compton, Newbury, Berkshire.
Entrican
G
G
Gelder
K I
KI
Collins
R A
RA
D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification*; D005822:Genetic Vectors; D066298:In Vitro Techniques; D007370:Interferon Type I / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D016898:Interferon-alpha / Q000235:genetics*; D007413:Intestinal Mucosa / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses; D011994:Recombinant Proteins; D012400:Rotavirus Infections / Q000235:genetics*; D012672:Semliki forest virus / Q000254:growth & development; D018411:Spodoptera; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D018076:DNA, Complementary; D007370:Interferon Type I; D016898:Interferon-alpha; D011994:Recombinant Proteins