Fig. 4.

BCCs induce CD73 expression in γδ1 T cells, which is dependent on exosomes and TGF-β1. a CD3+Vδ1 T cells sorted from PBHDs and co-cultured with BCCs (control or pretreated with 10 μm GW4869 for 24 h) were incubated in transwells for 24 h. CD73 expression in Vδ1 T cells was determined by flow cytometry. The right bar chart shows the proportion of the CD73+ subsets; data are shown as the means ± SD, n = 5, vs blank *p < 0.05, **p < 0.01. b CD3+Vδ1 T cells sorted from PBHDs were co-cultured with different BCCs in transwells or incubated with exosomes. The mRNA level of CD73 was evaluated by RT-PCR, n = 5. c Representative transmission electron micrograph of exosomes isolated from the supernatants of BC cell lines. d Western blot analysis of the exosome markers CD63 and CD9 in exosomes shed from different BC cell lines. e Exosomes isolated from the supernatants of BCCs (NC or pretreated with 10 μm GW4869 for 24 h), stained with PKH26, and then co-cultured for 24 h with Vδ1 T cells isolated from PBHDs. Then, the internalisation of exosomes was determined by flow cytometry. f CD3+Vδ1 T cells sorted from PBHDs and then co-cultured with BCCs or their exosomes (in the presence or absence of TGF-β1 or SB-431542 pretreatment) were incubated in transwells for 24 h. TGF-β1 treatment alone was performed as a control. CD73 expression in Vδ1 T cells was determined by flow cytometry. The right bar chart shows the CD73+ proportion of cell subsets. g CD3+Vδ1 T cells sorted from PBHDs and then co-cultured with BCCs or their exosomes (in the presence or absence of BMP4 or dorsomorphin pretreatment) were incubated in transwells for 24 h. BMP4 treatment alone was performed as a control. CD73 expression in Vδ1 T cells was determined by flow cytometry. The right bar chart shows the CD73+ proportion of cell subsets. Data are shown as the means ± SD, n = 5. vs MCF-10A, *p < 0.05, **p < 0.01, ***p < 0.001; vs TGF-β1 or BMP4, ^#p < 0.05. BCC breast cancer cell, PBHDs peripheral blood of healthy donors