Fig. 2

DOX uptake affects peripheral blood lymphocytes proliferation in vitro and ex vivo. A Representative proliferation profile of PBMC collected from healthy donors, labelled with CFSE 1 μM and stimulated with Concanavalin A (ConA, 0.5–5 μg/mL). Cells proliferation resulted in reduced CFSE mean fluorescence intensity (MFI). B Representative proliferation profile of PBMC collected from healthy donors, labelled with CFSE 1 μM, treated with DOX 5 μM for 24 h and stimulated with ConA (0.5–5 μg/mL). C Proliferation index of PBMC after 24 h in vitro treatment with DOX 5 μM calculated using the software FlowJo (version 10). Statistical significance: *p < 0.05 (paired t-test) (n = 3–6). D Proliferation index of CD4 + T cells after 24 h in vitro treatment with DOX 5 μM. Statistical significance: *p < 0.05 (paired t-test) (n = 3–6). (E) Proliferation index of CD8⁺ T cells after 24 h in vitro treatment with DOX 5 μM. Statistical significance: *p < 0.05 (paired t-test) (n = 3–6). F Representative profile of DOX MFI in PBMC collected from BC patient before and after the first cycle of DOX neoadjuvant chemotherapy. (G) DOX fluorescence signals detected by flow cytometry in matched PBMC from BC patients collected before and after the first cycle of DOX chemotherapy (n = 6). Statistical significance: ***p < 0.005 (paired t-test). H Representative profiles of CFSE MFI signal detected by flow cytometry before and 5 days after ConA stimulation in PBMC population collected from BC patients before and immediately after the first cycle of DOX NAC. I Proliferation index calculated at day 2, 3, 4 and 5 after ConA stimulation in matched PBMC collected before and immediately after the first cycle of DOX NAC (n = 6). Statistical significance: *p < 0.05, ***p < 0.005 (paired t-test)