A Virtual Infection Model Quantifies Innate Effector Mechanisms and Candida albicans Immune Escape in Human Blood

Candida albicans bloodstream infection is increasingly frequent and can result in disseminated candidiasis associated with high mortality rates. To analyze the innate immune response against C. albicans, fungal cells were added to human whole-blood samples. After inoculation, C. albicans started to filament and predominantly associate with neutrophils, whereas only a minority of fungal cells became attached to monocytes. While many parameters of host-pathogen interaction were accessible to direct experimental quantification in the whole-blood infection assay, others were not. To overcome these limitations, we generated a virtual infection model that allowed detailed and quantitative predictions on the dynamics of host-pathogen interaction. Experimental time-resolved data were simulated using a state-based modeling approach combined with the Monte Carlo method of simulated annealing to obtain quantitative predictions on a priori unknown transition rates and to identify the main axis of antifungal immunity. Results clearly demonstrated a predominant role of neutrophils, mediated by phagocytosis and intracellular killing as well as the release of antifungal effector molecules upon activation, resulting in extracellular fungicidal activity. Both mechanisms together account for almost of C. albicans killing, clearly proving that beside being present in larger numbers than other leukocytes, neutrophils functionally dominate the immune response against C. albicans in human blood. A fraction of C. albicans cells escaped phagocytosis and remained extracellular and viable for up to four hours. This immune escape was independent of filamentation and fungal activity and not linked to exhaustion or inactivation of innate immune cells. The occurrence of C. albicans cells being resistant against phagocytosis may account for the high proportion of dissemination in C. albicans bloodstream infection. Taken together, iterative experiment–model–experiment cycles allowed quantitative analyses of the interplay between host and pathogen in a complex environment like human blood.Author Summary: Candida albicans is the most important fungal pathogen in nosocomial bloodstream infections. So far little is known about the interplay of different cellular and non-cellular immune mechanisms mediating the protective response against C. albicans in blood. The in vivo scenario of C. albicans infection can be mimicked by human whole-blood infection assays to analyze the innate immune response against this pathogen. These experiments reveal the time-evolution of certain mechanisms while leaving the values of other quantities in the dark. To shed light on quantities that are not experimentally accessible, we exploited the descriptive and predictive power of mathematical models to estimate these parameters. The combination of experiment and theory enabled us to identify and quantify the main course of the immune response against C. albicans in human blood. We quantified the central role of neutrophils in the defence against this fungal pathogen, both directly by phagocytosis and indirectly by secreting antimicrobial factors inducing extracellular killing. Other findings include the distribution of C. albicans cells in neutrophils and monocytes as well as the immune escape of C. albicans cells in the course of infection.