01, compared with PBS). Our results indicate that the subunit immunogens HSP65-6 × P277 have been shown to be more effective than the immunogen containing only

HSP65 or P277 (*P < 0.05). To determine whether HSP65 serve as the carrier ZD6474 purchase may enhance the immunogenicity of P277, we analyzed Ab responses in HSP65-6 × P277-vaccinated animals. HSP65-6 × P277 protein showed greatly increased titers of anti-P277 antibodies by ELISA as early as 3 weeks following initial inoculation, while mice vaccinated with HSP65, P277 and PBS failed to elicit antibody formation. To identify the type of T cell that provided help for P277 antibody production, we characterized the isotype of the anti-P277 immunoglobins. The P277 antibodies in the HSP65-6 × P277 treated group were almost exclusively of the IgG1 and IgG2b subclass, which is indicative of Th2 help. In contrast, IgG2a P277 antibodies, which require Th1 help, were at very low levels in both the experimental and control groups (Fig. 1, *P < 0.05, compared with HSP65 and P277). These data suggest that Lapatinib in vivo the carrier HSP65 played a critical role in eliciting an immune response and enhancing

immunogenicity of the self-peptide P277 and nasal administration of HSP65-6 × P277 activated P277-specific Th2 response. At the end of the observation period, when the mice were 8 months

old, pancreata were obtained for histological examination. The predicament of the pancreas in mice that had been treated at 20 weeks showed a difference between the HSP65-6 × P277 treated and HSP65 or P277 treated mice: about 80% of islets in HSP65-6 × P277 treated mice but 40% of those in HSP65 and P277 treated mice were free of insuitis. The effectiveness of prevention insuitis of HSP65-6 × P277 is superior than the immunogen containing only HSP65 or P277 else (Fig. 2A). Fig. 2B depicts the results obtained on histological examination of the pancreas in the mice treated with HSP65-6 × P277: a significant increase in the number of islets free of insulitis, fewer necrosis areas formed in the pancreas tissue and a few lymphocytes filtrated around the islets of pancreas. From HSP65 or P277 vaccinated mice: a few necrosis areas formed in the pancreas tissue and a few lymphocytes filtrated around the islets of pancreas. In contrast, many necrosis and marked atrophy of pancreas islets showed and many lymphocytes filtrated around the islets in PBS-treated mice. We assayed the splenocytes isolated from HSP65-6 × P277, HSP65, P277 and PBS-treated animals to check their proliferative response to P277 and ConA. As shown in Fig.