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. Author manuscript; available in PMC: 2024 Sep 5.
Published in final edited form as: Cytotherapy. 2020 Dec 7;23(2):119–130. doi: 10.1016/j.jcyt.2020.11.002

Fig 3. Recognition of GXM from Cryptococcus spp. by GXMR-CAR+ T cells.

Fig 3.

(A) GXMR- CAR+ T cells and NoDNA T cells (mock-transduced, negative controls) were labeled with carboxyfluorescein succinimidyl ester and incubated with C. neoformans yeast labeled with Calcofluor-white. The interaction of GXMR-CAR+ T cells (green) and NoDNA T cells (green) with C. neoformans yeast (blue) was evaluated by using fluorescence microscopy. The bottom right image shows the cell clusters formed by co-localization of GXMR-CAR+ T cells and C. neoformans yeast. NoDNA T cells (top right) did not display co-localization. (B) GXMR- CAR+ and NoDNA T cells were assayed with polysaccharide antigens from the capsule of Cryptococcus spp., and the interaction of GXM with the cell surface was evaluated via flow cytometry by using an anti-GXM mAb 18B7 clone stained with a phycoerythrin-conjugated secondary antibody. The cells double-positive for 18B7 and GXMR-CAR-GFP demonstrated the interaction between GXMR-CAR+ T cells and GXM from Cryptococcus spp. The NoDNA T cells and GXMR-CAR+ T cells not incubated with polysaccharide antigens from the capsule of Cryptococcus spp. (without GXM) were used to demonstrate the absence of nonspecific binding of 18B7 to cells