Cryptococcus neoformans Secretes Small Molecules That Inhibit IL-1β Inflammasome-Dependent Secretion

Figure 4

1 kDa CM35 impacts phagocytic capacity and vomocytosis events in interactions between macrophages and C. neoformans. (a) Intracellular growth (CFU of 3 h vs. 24 h postinfection) of yeasts cells in BMMs stimulated with LPS (500 ng/ml) and H99 (2 : 1), with or without inhibitors (10% ). (b) Phagocytosis index (2 h postinfection) from BMMs stimulated with LPS (500 ng/ml) and H99 (5 : 1), with or without 1 kDa CM35 (10% ). (c–f) Flow cytometry analysis of (c, d) extracellular and (e, f) intracellular yeast cells (Calcofluor White high 2 h, 6 h, 12 h, and 24 h postinfection) from BMMs infected with H99 (10 : 1), with or without 1 kDa CM35 (10% ). (g) Scheme for transwell infection assay, illustrating the steps taken during the assay. (h, i) Flow cytometry measurement (24 h postinfection) of intracellular yeast cells in BMMs infected with H99 (2 : 1), in the upper chamber of a transwell apparatus. In the lower chambers, BMMs were stimulated with LPS (500 ng/ml) and B3501 or Δcap67 (5 : 1) (g). Alternatively, only yeast cells from B3501 or Δcap67 in a media with LPS (500 ng/ml) were included in the lower chambers (h). Statistical analysis was performed utilizing one-way ANOVA, where ns: not significant; ; ; . Comparisons were made with the B3501 lower chamber infected group (g).

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