Figure 1.

Th17 cells accumulate in locally invasive breast cancer. (A) PBMCs from HV or breast cancer patients (patients) were stained with anti-CD4⁺, anti-CD45RA, anti-CCR6, anti-CCR4, anti-CXCR3, anti-CD25 antibodies and analyzed by flow cytometry. The frequency of memory CD4⁺ (CD45RA⁻, CD4⁺) Th1 (CD25^low, CCR6⁻, CXCR3⁺), Th2 (CD25^low, CCR6⁻, CXCR3⁻, CCR4⁺), Th17 (CD25^low, CCR6⁺, CXCR3⁻), CD25^high Th17 (CD25^high, CCR6⁺, CXCR3⁻) and Treg (CD25^high, CCR6⁻, CXCR3⁻) cells is depicted. The data presented represent the analyses performed on 23 HVs and 13 breast cancer patients. (B) CD4⁺ lymphocytes from breast tumors (n =13) or normal breast tissue (n = 12) were analyzed as in (A). The frequency of memory CD4⁺ T cells is depicted. (C) Tumor infiltrating CD25^high Th17 cells and Treg cells were analyzed for IL-17 and RORγt expression by intracellular staining. Numbers beside outlined areas indicate percent cells in gate. (D) Tumor infiltrating CD25^high Th17 cells and Treg cells were analyzed for Foxp3 expression by intracellular staining. Numbers beside outlined areas indicate percent cells in gate. (E) Tumor infiltrating CD25^high Th17 and Treg cells sorted from breast tumors were restimulated with anti-CD3 and anti-CD28 antibodies and IL-17A secretion was assessed by ELISA after 3 d. Representative data from one of at least three independent experiments are shown (C–F). *p < 0.05