title: Amended recombinant cells (ARCs) expressing bovine IFN-gamma: an economical and highly effective adjuvant system.
abstract: Interferon-gamma (IFN-gamma) has potent immune stimulatory activity, but purifying recombinant protein is expensive, soluble cytokine delivery is inefficient, and high doses of IFN-gamma cause adverse systemic effects. We've shown, however, that chemically fixed, amended recombinant Pseudomonas fluorescens cells (ARCs), expressing soluble bovine IFN-gamma (BGI/ARCs), provide an effective production and delivery vehicle for highly active cytokine. In this report we investigate the immune enhancing activity of BGI/ARCs in the presence or absence of two other adjuvants which enhance cell recruitment and protein uptake. Adjuvant activity and immune-mediated protection was evaluated using recombinant, truncated glycoprotein-D (tgD) and a bovine herpesvirus-1 (BHV-1) disease challenge. Our initial dose-titration study showed that 100 microg of recombinant IFN-gamma in BGI/ARCs significantly increased both IgG1 and IgG2 tgD-specific antibody titres and IgG1/IgG2 ratios were significantly reduced with as little as 1.0 microg IFN-gamma. Vaccine formulation studies, using 20 microg tgD/vaccine dose and 100 microg IFN-gamma delivered in BGI/ARCs, formulated in phosphate buffered saline (PBS), induced significantly increased antibody responses, following both primary and secondary immunization, and immune-mediated protection following a BHV-1 respiratory infection. Co-formulating BGI/ARCs with either an oil and water emulsion (Emulsigen) or a polyphosphazene polymer (PCPP) did not significantly enhance tgD-specific antibody titres or disease protection when compared with BGI/ARCs alone. Surprisingly, co-delivering a single dose of BGI/ARCs with tgD protein in PBS had optimal adjuvant activity and the dose of IFN-gamma delivered was four-fold less than the dose previously shown to induce adverse systemic responses.
authors.affiliation: The Dow Chemical Company, 5501 Oberlin Dr., San Diego, CA 92121, USA.
authors.forename: Frank H
authors.initials: FH
authors.lastname: Gaertner
authors.forename: Lorne A
authors.initials: LA
authors.lastname: Babiuk
authors.forename: George
authors.initials: G
authors.lastname: Mutwiri
authors.forename: Janna M
authors.initials: JM
authors.lastname: Armstrong
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Griebel
mesh_terms: D000276:Adjuvants, Immunologic / Q000235:genetics / Q000627:therapeutic use; D000818:Animals; D000917:Antibody Formation; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology; D002986:Clinical Trials as Topic; D005260:Female; D006566:Herpesviridae Infections / Q000276:immunology / Q000662:veterinary; D017589:Immunization Programs; D007371:Interferon-gamma / Q000235:genetics*; D008297:Male; D011994:Recombinant Proteins / Q000276:immunology / Q000627:therapeutic use; D011995:Recombination, Genetic; D018140:Varicellovirus / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D011994:Recombinant Proteins; D007371:Interferon-gamma