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3A). Interestingly, when the TLR-9 ligand CpGB ( Fig. 3B) but not the TLR-3 ligand Poly I:C (data not shown) was co-adsorbed with TT to YC-Brij700-chitosan NP, the T-cell proliferation response was further enhanced (P?[http://www.selleckchem.com/products/wortmannin.html selleck chemical] ( Fig. 3B). In addition, although CpGB induced T-cell proliferation on its own, this induction was significantly lower than that induced by TT-CpGB co-adsorbed NP. Further confirmation of the enhancing effect on T-cell [http://en.wikipedia.org/wiki/Thalidomide Thalidomide] proliferation by co-adsorption of TT plus CpGB on NP, was demonstrated when instead of using TT, the irrelevant Ag BSA was co-adsorbed to NP with CpGB ( Fig. 3B). To test whether NP could enhance T-cell proliferative responses to gp-140, splenocytes from gp140-immunized mice were used in vitro. Splenocytes were cultured in the presence of Ag alone or gp140-adsorbed NP and the incorporation of 3H[Td] into DNA measured after three days of culture. gp140-adsorbed NP but not naked NP enhanced splenocyte proliferative responses to gp140 (P?[http://www.selleckchem.com/mTOR.html mTOR inhibitor] and IgA, animals were immunized i.d. with gp140 following a prime-boost-boost protocol at 30 day intervals. Serum samples were taken before each immunization and 30 days after the last boost, and the levels of IgG and IgA were tested by gp140-specific ELISA. gp140 alone induced significant levels of IgG but these levels were much higher when the Ag was adsorbed to NP (Fig. 4B). Such IgG levels were comparable to those induced by Alum (day 60), and differences were already observable following a single prime (day 30). Plateau IgG levels were already observed after first boost (day 60, Fig. 4B).