(A) Quantification of spontaneous and organoid-induced IFNγ production by CD4+ T cells obtained by two week co-culture with autologous tumor organoids. Error bars indicate s.e.m. of at least 2 biological replicates.
(B) Quantification of CD137 expression by CD4+ T cells upon re-stimulation with the tumor organoids used for induction, normal colon organoids, or organoids of a mismatch repair proficient (pMMR) CRC from the same patient. T cells were obtained by two week co-culture with autologous tumor organoids and where indicated (lower case e) further expanded using a rapid expansion protocol (Dudley et al. 2013), in some cases preceded by CD137-based enrichment of tumor-reactive cells (indicated by upper case E). Error bars indicate s.e.m. of at least 2 biological replicates.
(C) Quantification of CD137 expression by CD4+ T cells after stimulation with unloaded or Geltrex-loaded monocyte-derived dendritic cells (DCs), irradiated tumor cells, or organoids cultured with or without Geltrex. Error bars indicate s.d. of technical replicates.
See also Figure S4.