Conventional amphotericin B elicits markers of immunogenic cell death on leukemic blasts, mediates immunostimulatory effects on phagocytic cells, and synergizes with PD-L1 blockade.

Immunostimulatory regimens are a game changer in the fight against cancer, but still only a minority of patients achieve clinical benefit. Combination with immunomodulatory drugs and agents converting otherwise non-immunogenic forms of cell death into bona fide "immunogenic cell death" (ICD) could improve the efficacy of these novel therapies. The aim of our study was to investigate conventional Amphotericin B (AmB) as an enhancer of antitumor immune responses. In tumor cell line models, AmB induced ICD with its typical hallmarks of calreticulin (CALR) expression and release of high mobility group box 1 (HMGB1) as well as Adenosine 5'-triphosphate (ATP). Interestingly, in contrast to non-ICD inducing treatments, ICD induction led to up-regulation of PD-L1-expression by ICD experiencing cells, resulting in decreased maturation of dendritic cells (DCs). Blocking this PD-L1 expression on tumor cells could unleash full ICD effects on antigen presenting cells. Even at sub-toxic concentrations, AmB was able to enhance CALR on leukemic blasts, particularly on phagocytic monoblastic THP-1 cells, which also showed features of "M1-like" differentiation after AmB exposure. The ability of AmB to increase the immunogenicity of tumor cells was confirmed in vivo in a mouse vaccination experiment. In conclusion, we demonstrate that AmB can promote antitumor immune responses in a dose-dependent manner by ICD induction, surface translocation of CALR on leukemic blasts even at sub-toxic concentrations, and "M1-like" polarization of phagocytic cells, making it noteworthy as potential booster for cancer immunotherapy. We additionally report for the first time that PD-L1 expression may be a feature of ICD, possibly as a negative feedback mechanism regulating the maturation status of DCs and thus indirectly affecting T-cell priming.

Effect of different sample volumes on the DNA extraction of Aspergillus fumigatus from whole blood.

Five methods were compared, using conventional PCR, for the isolation of DNA from Aspergillus fumigatus conidia from 1-3-mL samples of whole blood. A lower detection threshold of Aspergillus conidia was achieved using 3-mL rather than 1-mL samples with three of five methods tested.

Paradoxical growth effects of the echinocandins caspofungin and micafungin, but not of anidulafungin, on clinical isolates of Candida albicans and C. dubliniensis.

OBJECTIVES: To analyze the effects of a high concentration of three antifungal substances, the echinocandins anidulafungin, caspofungin, and micafungin, on the growth of Candida spp. METHODS: The growth of 127 C. dubliniensis isolates and 103 C. albicans isolates cultured in medium containing anidulafungin, caspofungin, or micafungin was analyzed using a broth microdilution test according to the guidelines of the CLSI M27-A2 [NCCLS (1997), Wayne, PA]. The final concentrations of all three echinocandins ranged from 0.125 to 64 microg/L. RESULTS: The different effects of these three antifungal substances on C. albicans cells in comparison to C. dubliniensis cells were quite distinct. When both Candida species were grown in the presence of anidulafungin only a trailing effect was observed. Micafungin induced an Eagle effect in C. dubliniensis only (63%), while caspofungin induced this effect in the majority of C. dubliniensis isolates (90%) and in only a few C. albicans isolates (14%). CONCLUSIONS: Based on our observations, anidulafungin has effects that are different from the ones produced by micafungin and caspofungin. Whether this different response to high concentrations of echinocandins is based on genetic or phenotypic differences between C. albicans and C. dubliniensis has to be determined in future experiments.