Janna M Armstrong

Dow AgroSciences, Indianapolis, Indiana, United States

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Publications (2)11.13 Total impact

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    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.
    Vaccine 02/2009; 27(9):1377-85. · 3.49 Impact Factor
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    ABSTRACT: Recombinant Pseudomonas fluorescens cells, expressing over 40% protein as bovine interferon-gamma (IFN-gamma), were chemically fixed to sterilize the culture and amend the bacterial cell wall. When killed and fixed recombinant cells, termed here amended-recombinant-cells (ARCs), were assayed for interferon activity, we obtained the following surprising results: 1) sterilization and fixation did not inactivate ARC-encapsulated IFN-gamma; 2) ARC-encapsulated IFN-gamma and soluble, recombinant IFN-gamma were equally active in vitro but proteolysis was required for release of the ARC cytokine; and 3) ARC-encapsulated IFN-gamma was active in vivo with optimal adjuvant activity at a dose about 1000-fold less than previously reported for soluble, recombinant IFN-gamma and 100-fold less than doses which induced adverse systemic effects. The mechanism by which ARC-encapsulation increased IFN-gamma activity in vivo remains uncertain. However, our in vitro results show that sustained release of soluble IFN-gamma is a likely factor. The ARC production and delivery system achieves enhanced adjuvant activity with reduced risk of systemic effects, and the low cost of IFN-gamma production offers new opportunities for the use of this important cytokine.
    Journal of Controlled Release 11/2005; 107(2):189-202. · 7.63 Impact Factor