Skip to main content
. 2022 Oct 27;13(2):410–431. doi: 10.1158/2159-8290.CD-22-0523

Figure 5.

Figure 5. Senescent cancer cells from human patients hyperstimulate autologous reactive TILs. A, Schematics of the procedure for isolating, amplifying, classifying, and coculturing patient-derived tumor cells with autologous reactive and nonreactive TILs (left). Table indicating patients and corresponding tumor type used in this study (right). B, Flow cytometry analysis of 4-1BB activation marker in CD8 cells from nonreactive (F1 and F2 fragments) and reactive (F3 and F4 fragments) autologous TILs from VHIO-008 patient after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-008 cells, or bleomycin-treated senescent VHIO-008 cells (as indicated). C, Flow cytometry analysis of 4-1BB activation marker in CD8 cells from nonreactive (F1 fragment) and reactive (F3 fragment) autologous TILs from patient VHIO-009 after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-009 cells, or bleomycin-treated senescent VHIO-009 cells (as indicated). D, Flow cytometry analysis of the 4-1BB activation marker in CD8 cells from nonreactive (F1 fragment) and reactive (F2 and F3 fragment) autologous TILs from patient VHIO-088 after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-088 cells, or bleomycin-treated senescent VHIO-088 cells (as indicated). SSC-A, side scatter area. E, Flow cytometry analysis of the 4-1BB activation marker in nonreactive PBL CD8 T cells and CD8 cells from reactive (F1, F2, and F3 fragments) autologous TILs from patient VHIO-35035 (abbreviated as V-35035) after culture in RPMI medium either alone or with anti-CD3 (OKT3), control V-35035 cells, or bleomycin-treated senescent V-35035 cells (as indicated). F, Flow cytometry analysis of 4-1BB activation marker in CD8 cells from reactive autologous TILs (reactive F3, F4 and F5 fragments) enriched to be reactive against MAGEB2p.E167Q and RPL14p.H20Y (two neoantigens previously identified by whole-exome sequencing of the autologous tumor cell line) after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-008 cells, or bleomycin-treated senescent VHIO-008 cells (as indicated).

Senescent cancer cells from human patients hyperstimulate autologous reactive TILs. A, Schematics of the procedure for isolating, amplifying, classifying, and coculturing patient-derived tumor cells with autologous reactive and nonreactive TILs (left). Table indicating patients and corresponding tumor type used in this study (right). B, Flow cytometry analysis of 4-1BB activation marker in CD8 cells from nonreactive (F1 and F2 fragments) and reactive (F3 and F4 fragments) autologous TILs from VHIO-008 patient after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-008 cells, or bleomycin-treated senescent VHIO-008 cells (as indicated). C, Flow cytometry analysis of 4-1BB activation marker in CD8 cells from nonreactive (F1 fragment) and reactive (F3 fragment) autologous TILs from patient VHIO-009 after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-009 cells, or bleomycin-treated senescent VHIO-009 cells (as indicated). D, Flow cytometry analysis of the 4-1BB activation marker in CD8 cells from nonreactive (F1 fragment) and reactive (F2 and F3 fragment) autologous TILs from patient VHIO-088 after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-088 cells, or bleomycin-treated senescent VHIO-088 cells (as indicated). SSC-A, side scatter area. E, Flow cytometry analysis of the 4-1BB activation marker in nonreactive PBL CD8 T cells and CD8 cells from reactive (F1, F2, and F3 fragments) autologous TILs from patient VHIO-35035 (abbreviated as V-35035) after culture in RPMI medium either alone or with anti-CD3 (OKT3), control V-35035 cells, or bleomycin-treated senescent V-35035 cells (as indicated). F, Flow cytometry analysis of 4-1BB activation marker in CD8 cells from reactive autologous TILs (reactive F3, F4 and F5 fragments) enriched to be reactive against MAGEB2p.E167Q and RPL14p.H20Y (two neoantigens previously identified by whole-exome sequencing of the autologous tumor cell line) after culture in RPMI medium either alone or with anti-CD3 (OKT3), control VHIO-008 cells, or bleomycin-treated senescent VHIO-008 cells (as indicated).