Each mouse was treated with 50?ï¿½ï¿½g of liposomized-tuftsin intraperitoneally for three consecutive days as described earlier (Khan et?al., this website 2003). Pretreatment with tuftsin Liposomes (PT-tuftsin-liposomes) in leukopenic Mice was started 8?h after cyclophosphamide injection. After 3?days of tuftsin treatment, each mouse was infected with 5?ï¿½ï¿½?105 cells of C.?neoformans. Mice were divided into the following groups: (1) saline, (2) sham liposomes, (2) PT-tuftsin-liposomes?+?sham liposomes, (4) tuftsin liposomes, (5) PT-tuftsin-liposomes?+ tuftsin liposomes, (6) liposomal nystatin (5?mg?kg?1), (7) PT-tuftsin-liposomes?+?liposomal nystatin (5?mg?kg?1), (8) tuftsin-bearing liposomal nystatin (5?mg?kg?1), (9) PT-tuftsin-liposomes?+?tuftsin-liposomal nystatin (5?mg?kg?1). The prophylactic and therapeutic role of tuftsin was evaluated by determining the fungal load in brain tissues. Three mice from each group were sacrificed on day 3 post-C.?neoformans infection (8?h after the 2nd dose of treatment) and their brains were taken out aseptically as described earlier (Nasti et?al., 2006). Briefly, weighed portions of the brain tissues were homogenized in 5?mL of sterile normal saline and different dilutions of the suspension were plated on SD agar plates containing chloramphenicol. The plates were incubated at 37?ï¿½ï¿½C for 48ï¿½C72?h. The numbers of colonies were counted and the fungal load was determined by multiplying them by the dilution factor. Analysis of the survival of mice was Ibrutinib performed using Kaplanï¿½CMeier curve, and various groups were compared by log-rank test. Fungal burden in organs was analyzed by one-way anova followed by the Bonferroni post hoc test using graphpad prism software version 3.0. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of nystatin at which there was complete inhibition of the fungal growth. The MIC of nystatin for C.?neoformans was found to be 1.5?ï¿½ï¿½g?mL?1. Immuno-potential effect of tuftsin was analyzed by treating leukopenic mice with tuftsin-bearing liposomes, and mice treated with saline or sham liposomes acted as controls. The number of leukocytes was counted in the blood of tuftsin-treated Akt inhibitor or untreated mice. Our results demonstrated that treatment with tuftsin liposomes induced early recovery of leukocytes in leukopenic mice. We found that leukopenic mice treated with tuftsin-liposomes have higher counts of leukocytes (1220ï¿½C1956) on day 4 post-cyclophosphamide injection compared with mice treated with saline (166ï¿½C272) or sham liposomes (183ï¿½C344) (P?0.001). Moreover, the group of leukopenic mice treated with tuftsin-incorporated liposomes showed complete recovery of leukocytes on day 8 post-cyclophosphamide treatment, whereas saline- or sham liposome-treated mice showed only 40ï¿½C60% recovery in leukocyte counts. Cryptococcus neoformans can multiply inside macrophages as shown by an increase in the numbers of CFUs after 24?h of infection (Fig.