Figure 5

CD4⁺LAP⁺ tumor-infiltrating lymphocytes (TILs) produce immunosuppressive cytokines. Intracellular cytokine staining of CD4⁺LAP^+/− T cells, with and without phorbol 12-myristate 13-acetate (PMA)/ionomycin stimulation, from three matched (a) peripheral blood mononuclear cell (PBMC), (b) unaffected colon, and (c) tumor samples (2 Dukes’ B, 1 Dukes’ C). Representative fluorescence-activated cell sorting (FACS) plots for interleukin (IL)-17A, interferon-γ (IFN-γ), and IL-10 are shown and significant differences are indicated: *P<0.05 and **P<0.01. (d) Representative staining for active transforming growth factor-β (TGF-β) expression on unstimulated CD4⁺LAP⁺ TILs (black line) and CD4⁺LAP⁻ TILs (gray shading) isolated from a Dukes’ C tumor. (e) CD4⁺LAP⁺ TILs (black line), isolated from a Dukes’ C tumor, were stained with PKH-26 and stimulated with αCD3/28 beads; proliferation in vitro was measured over 72 h. Gray shading indicates the PKH-26-positive control (unstimulated effector T cells). LAP, latency-associated peptide.

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