title: Murine interferon-gamma/interleukin-1 fusion proteins used as antigens for the generation of hybridomas producing monoclonal anti-interleukin-1 antibodies.
abstract: In several biological systems interferon-gamma (IFN-gamma) and interleukin-1 (IL-1) act synergistically. We therefore examined whether it would be possible to construct IFN-gamma/IL-1 hybrid proteins that would be more active than the individual components. Hybrid proteins were examined that consisted of the amino-terminal 118 residues of mouse IFN-gamma and the 156 or 152 carboxyl-terminal residues of mouse IL-1 alpha or IL-1 beta, respectively. They were obtained by ligation of the respective coding sequences and expression of the fused genes under control of the PL promotor in Escherichia coli. Both the IFN-gamma/IL-1 alpha and the IFN-gamma/IL-1 beta fusion proteins were purified by affinity chromatography on an anti-IFN-gamma monoclonal antibody column. Analysis of biological activities showed that these fusion proteins were less active than the individual cytokines. Specific antiviral activity of the IFN-gamma/IL-1 beta hybrids was less than 0.1% that of IFN-gamma and D10.G4.1 T-cell proliferative (IL-1) activity amounted to 0.1% that of mouse IL-1. Affinity-purified preparations of the IFN-gamma/IL-1 alpha hybrid were found to contain variable proportions of a Mr 14,000 degradation product possessing IFN-gamma activity, whereas the undegraded Mr 30,000 fusion protein, while being devoid of detectable IFN-gamma activity, did possess IL-1 activity (1%). Serum from rats immunized with the IFN-gamma/IL-1 alpha hybrid contained high levels of IL-1 alpha-binding and -neutralizing antibodies and IFN-gamma-binding antibodies, but no detectable levels of IFN-gamma-neutralizing antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Rega Institute, K.U. Leuven, Belgium.
authors.forename: R
authors.initials: R
authors.lastname: Dijkmans
authors.forename: E
authors.initials: E
authors.lastname: Martens
authors.forename: E
authors.initials: E
authors.lastname: Beuken
authors.forename: F
authors.initials: F
authors.lastname: Cornette
authors.forename: C
authors.initials: C
authors.lastname: Dillen
authors.forename: H
authors.initials: H
authors.lastname: Heremans
authors.forename: D
authors.initials: D
authors.lastname: Boraschi
authors.forename: A
authors.initials: A
authors.lastname: Billiau
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D001681:Biological Assay; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006825:Hybridomas / Q000276:immunology; D007118:Immunoassay; D007371:Interferon-gamma / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D007375:Interleukin-1 / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology; D008213:Lymphocyte Activation; D051379:Mice; D010957:Plasmids; D051381:Rats; D011919:Rats, Inbred Strains; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D007375:Interleukin-1; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D007371:Interferon-gamma