Abstract

A WATER-SOLUBLE CHITOSAN AS A CARRIERS FOR DNA VACCINE: THE NANOCOMPLEXES CHARACTERIZATION AND IN VIVO TRANSFECTIONS STUDIES

Rosa V. S. Amorim*, Rodrigo P. Pedrosa, Veruska F Alves, Eduardo J. M. Nascimento and Norma Lucena-Silva

ABSTRACT

The efficacy of water-soluble chitosan (WSCh) acting as a carrier to a DNA vaccine model, was evaluated here, using a plasmid DNA, encoding a green fluorescent protein (pGFP). WSCh-pGFP nanocomplexeses were prepared using an easy complex coacervation method, in different WSCh-pGFP mass ratio. The size and morphology of complexes, protectively from DNase I and the WSCh-pGFP nanocomplexes expression in vivo were determined, by inoculating 50 g of pGFP naked or associated with chitosan, in mass ratios of 4, and then injected subcutaneously in female BALB/c mice. The nanocomplexes formation was confirmed by Electrophoresis analysis on a 1% agarose gel and Trasmission Eletron Microscopy (TEM). The analysis of the results showed that WSCh-pGFP, ratio 4:1 (wt/wt) showed whole retention of DNA migration and DNase I protection from 30min at least. The WSCh-pGFP nanocomplexess had morphology like a ball-shaped, ranged from 50 to 200 nm in size andWSCh-pGFP nanocomplexes expression was confirmed, in vivo, in epidermis as lymphoid organs up to 35 days. by Fluorescent Microscopy.

Keywords: DNA vaccine; gene delivery; glycol-chitosan; cell transfection.