Parasite persistence and concomitant immunity were achieved by Lm/CpG 10, 11 in the absence of lesions. In order to understand and exploit the immunological features of Lm/CpG, we have continued to unravel how the immune response Lumacaftor cost to this vaccine is different from natural infection (leishmanization). We have discovered that Lm/CpG promotes the early proliferation of dermal Th17 cells, contrasting with the highly polarized Th1 response that takes place much later in mice vaccinated

with L. major alone. Most importantly, Th17 cells appear to be the predominant effector population in Lm/CpG-vaccinated mice, although Th1 cells are also present. Neutralization of IL-17 (confirmed by the use of IL-17 receptor-deficient mice) causes enhanced susceptibility to L. major infection (higher parasite burdens, development of lesions), accompanied by a decrease in IFN-γ production, in neutrophil migration, and by an increase in Treg frequencies. The intradermal model of infection produces an immunologically “silent”

phase during the first 2–3 wk 13, 14. We have reported that the combination of live parasites and CpG DNA eliminates such a phase by causing a Adriamycin order rapid activation of DC, release of proinflammatory cytokines, and migration of activated lymphocytes to the vaccine site 10, 11. We obtained a full cytokine profile of the vaccination site of mice immunized with the live vaccines (L. major or leishmanization versus Lm/CpG) or with CpG DNA alone as a control. We extracted cells from the dermis of vaccinated animals prior to vaccination (wk 1), and 2 wk (“silent” phase for L. major, activated phase for Lm/CpG), 6 wk (acute phase for L. major), and 10 wk post vaccination (chronic phase). Cells were restimulated ex vivo with the vaccines to determine the production of various cytokines in the culture supernatants. As shown in Table 1, we found significant differences in the time frame of the immune response among the experimental groups. Cytokines Baf-A1 in vitro were secreted at low levels in the uninfected skin (wk 1). As reported by us 10, 11, IL-6 production was significantly increased during the “silent” phase (wk 2)

in Lm/CpG-vaccinated mice. IL-12, TNF-α, IL-17, and IFN-γ were elevated at the same time point, confirming the early proinflammatory response initiated by Lm/CpG vaccination. TGF-β secretion was slightly elevated in the Lm/CpG when compared with L. major alone, although it was very low. Conversely, IL-10 secretion was lower in the ears of the Lm/CpG-vaccinated mice at this time point; again, the overall values were close to the limit of detection. Although IL- 4 secretion was higher in the Lm/CpG-vaccinated animals at wk 2, its level was very low at all time points and all groups, as expected from the genetically resistant C57BL/6 mouse. Wk 6 values revealed a reversal in cytokine profiles, with the L. major-vaccinated animals now showing a proinflammatory response significantly dominated by the production of IL-12 and IFN-γ.