Conservation of intracellular pathogenic strategy among distantly related Cryptococcal species

ABSTRACT: We compared the interaction between Cryptococcus gattii species complex strain R265 and Cryptococcus neoformans species complex strain H99, molecular type VNI, with murine macrophages to ascertain similarities and differences in their intracellular pathogenic strategy. Parameters analyzed include non-lytic exocytosis using time-lapse microscopy, phagolysosomal pH by dual fluorescent microscopy, intracellular capsular growth, phagolysosomal membrane permeabilization by flow cytometry, and the macrophage transcriptional response by gene expression microarray analysis. Using microarrays, we compared the transcriptional response of Bone Marrow-derived macrophages (BMDM) infected with C. gattii species complex strain R265 or C. neoformans species complex strain H99, relative to uninfected BMDM collected from the same time intervals, 2hr and 24 hr post-infection. For analysis following normalization and summarization steps, a 2-way Analysis of Variance (ANOVA) with linear contrasts for treatment (infection/strain) and time (2 h or 24 h) vs. Control (uninfected) was performed with outputs of P value, Fold Change, and Mean Ratio. A linear contrast of uninfected controls (24 h uninfected vs 2 h uninfected) was also performed to generate a ‘time only’ gene list. Cutoff criteria for filtering gene lists was significant P values (p<0.01) with fold changes greater that 2 or less than –2. Ten genes from the filtered gene lists were selected for validation by qPCR, with four housekeeping genes for normalization and comparison.

ORGANISM(S): Mus musculus

PROVIDER: GSE112668 | GEO | 2018/06/30

REPOSITORIES: GEO