Fig 1: Jurkat cell activation mediated by GXMR-CAR constructs in the presence of Cryptococcus spp. yeasts was validated by expression of the activation marker CD69. Jurkat cells (2 × 105 cells/mL) expressing GXMR-8-BB?, GXMR-8-28? or Lenti-mock constructs, after transduction using an MOI of 3, were incubated with heat-killed yeast of C. gattii or C. neoformans (1:1 ratio of cells:yeast) or only medium for 20 h of culture. After this period of incubation, the cells were stained with a human anti-CD69 APC-conjugated antibody or isotype control, and the CD69 expression was measured by flow cytometry. (a) Representative histograms of CD69 expression and (b) quantification of the median of the CD69-APC fluorescence intensity after 20 h of co-culture between transduced cells and heat-killed yeast or only medium. The values are expressed as mean ± SD; and *, # p < 0.05, **, ## p < 0.01 according to one-way ANOVA followed by Tukey’s multiple comparisons test. # denotes statistical significance when GXMR-CAR constructs or Lenti-mock in the presence of heat-killed yeasts were compared to medium, whereas * represents a significant difference between GXMR-CAR and Lenti-mock in the same condition.