Journal
JOURNAL OF CONTROLLED RELEASE
Volume 161, Issue 1, Pages 116-123Publisher
ELSEVIER
DOI: 10.1016/j.jconrel.2012.04.015
Keywords
Avian influenza virus; H5 DNA vaccine; HI antibody; T cell; Cytokine; Green synthesis AgNP
Funding
- Agro-Biotechnology Institute, Ministry of Science, Technology and Innovations [06-05-ABI-AB002]
- HICOE grant, Ministry of Higher Education, Government of Malaysia
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DNA formulations provide the basis for safe and cost effective vaccine. Low efficiency is often observed in the delivery of DNA vaccines. In order to assess a new strategy for oral DNA vaccine formulation and delivery, plasmid encoding hemagglutinin (HA) gene of avian influenza virus, A/Ck/Malaysia/5858/04 (H5N1) (pcDNA3.1/H5) was formulated using green synthesis of sliver nanoparticles (AgNP) with polyethylene glycol (PEG). AgNP were successfully synthesized uniformly dispersed with size in the range of 4 to 18 nm with an average size of 11 nm. Cytotoxicity of the prepared AgNP was investigated in vitro and in vivo using MCF-7 cells and cytokine expression, respectively. At the concentration of -5 log(10)AgNP, no cytotoxic effects were detected in MCF-7 cells with 9.5% cell death compared to the control. One-day-old specific pathogen-free (SPF) chicks immunized once by oral gavage with 10 mu l of pcDNA3.1/H5 (200 ng/ml) nanoencapsulated with 40 mu l AgNP (3.7x10(-2) mu g of Ag) showed no clinical manifestations. PCR successfully detect the AgNP/H5 plasmid from the duodenum of the inoculated chicken as early as 1 h post-immunization. Immunization of chickens with AgNP/H5 enhanced both pro inflammatory and Th1-like expressions, although no significant differences were recorded in the chickens inoculated with AgNP, AgNP/pcDNA3.1 and the control. In addition, serum samples collected from immunized chickens with AgNP/H5 showed rapidly increasing antibody against H5 on day 14 after immunization. The highest average antibody titres were detected on day 35 post-immunization at 51.2 +/- 7.5. AgNP/H5 also elicited both CD4+ and CD8+ T cells in the immunized chickens as early as day 14 after immunization, at 7.5 +/- 2.0 and 20 +/- 1.9 percentage, respectively. Hence, single oral administrations of AgNP/H5 led to induce both the antibody and cell-mediated immune responses as well as enhanced cytokine production. (C) 2012 Elsevier B.V. All rights reserved.
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