Figure 2

Effect of chitosan interferon-γ nanogene (CIN) therapy on expression of dendritic cell activation markers. A, In vitro CIN treatment of CD11c⁺ cells from the lungs of ovalbumin (OVA)-allergic mice. BALB/c mice were OVA sensitized and challenged with OVA and sacrificed 18 hours later. Lungs were removed and CD11c⁺ dendritic cells were isolated as described in Materials and Methods. Dendritic cells were cultured with vector alone (control) or with CIN. Flow cytometry was performed on cell suspensions after labelling for CD11c (fluorescein isothiocyanate [FITC]) and for each of the following markers: I-Ad, CD40, and CD80 phycoerythrin (PE). Counts were done using a FACScan gated for CD11c cells. The figure shows the percentage of cells that were positive for CD11c and for each of the markers. Activation marker expression in CD11c⁺ cells from the bronchoalveolar lavage (BAL) fluid and lymph nodes of OVA-allergic mice treated with CIN (B). Mice were treated as described in Materials and Methods, and CD11c⁺ cells were isolated from BAL fluid and lymph nodes. Cells were analyzed by flow cytometry for the expression of CD40, CD80, CD86, and I-Ad gated to CD11c⁺b⁺. The data are based on cytometry of a minimum of 15,000 cells and were substantiated by a repeat experiment.