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. 2019 Dec 27;9:20068. doi: 10.1038/s41598-019-56670-x

Figure 5.

Figure 5

SPN limits tumor cell clustering with T cells. (A) To detect T cell-tumor cell clusters, healthy donor T cells were labeled with CellTrace CFSE and tumor cells were labeled with CellTrace Violet (CTV) or CellTrace Yellow (CTY). Differentially labeled tumor cells were mixed at a 1:1 ratio and co-cultured with labeled T cells (3:1 E:T) + 30 ng/ml CD20xCD3 bsAb or CD3-binding isotype control antibody. After 18 hours, cells were collected, fixed in 4% paraformaldehyde and immediately analyzed by FACS. (A) Representative plots showing the detection of T cell-tumor cell clusters. JeKo-1/NT cells in T cell conjugates are CTV + FITC +, and JeKo-1/SPN cells in T cell conjugates are CTY + FITC+ . (C) The average percent of tumor cells in conjugates with T cells was quantified across triplicate samples. Note that the extent of clustering was not affected by CellTrace dyes. ***P < 0.001 by two-tailed T-test. (D) Representative plots showing the detection of T cell-tumor cell clusters. JeKo-1/NT cells in T cell conjugates are CTV + FITC +, and JeKo-1/CD52 cells in T cell conjugates are CTY + FITC +. (E) The average percent of tumor cells in conjugates with T cells was quantified across triplicate samples. Note that the extent of clustering was not affected by CellTrace dyes. ***P < 0.001 by two-tailed T-test.