Poly-β amino ester-containing microparticles enhance the activity of nonviral genetic vaccines

Little et al. 10.1073/pnas.0403549101.

Supporting Information

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Supporting Figure 6
Supporting Figure 7
Supporting Movie 1
Supporting Movie 2





A


B

Fig. 6. Microparticles prepared from poly lactic-co-glycolic acid (PLGA) (A) and 25% poly-b amino ester (PBAE) (B) show high surface integrity as determined by scanning electron microscopy. Magnification is ´ 1,000 (Inset, ´ 5,000).





Fig. 7. In vitro activation of T cells using primary bone marrow-derived dendritic cells treated with PBAE microparticle formulations. A total of 2 × 104 L3100, SIY antigen-specific T cells (1) were added to wells containing 2 × 105 (10:1, filled bars) and 1 × 105 (5:1, open bars) dendritic cells treated with microparticle formulations encapsulating pCMV-SIY plasmid DNA for 24 h. Activation was measured by transferring the T cells to an IFN-genzyme-linked immunospot plate after 18 h of incubation with the dendritic cells, and the plate was processed and analyzed the next day for number of IFN-gspots. Positive controls were performed (second column) with 10-6 M SIY peptide incubated with the dendritic cells 4 h before L3100 addition. Negative controls were untreated dendritic cells (first column) and 25% PBAE microparticles containing pCMV-Luc (sixth column). The results shown are averages of four repeats, and standard error bars are included. Asterisks indicate a significant difference from the negative control at the same DC:T cell ratio using comparative ANOVA.





Supporting Movie 1

Movie 1. Full 3D fluorescence movie of primary human peripheral blood mononuclear cell-derived dendritic cells (DCs) treated with PLGA and represented as an overhead image in Fig. 2.





Supporting Movie 2

Movie 2. Full 3D fluorescence movies of primary human peripheral blood mononuclear cell-derived dendritic cells (DCs) treated with 25% PBAE microparticle formulations and represented as an overhead image in Fig. 2.

1. Sykulev, Y., Vugmeyster, Y., Brunmark, A., Ploegh, H. L. & Eisen, H. N. (1998) Immunity 9, 475-483.

This Article

  1. Published online before print June 21, 2004, doi: 10.1073/pnas.0403549101 PNAS June 29, 2004 vol. 101 no. 26 9534-9539
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