Targeting breast cancer stem cells by dendritic cell vaccination in humanized mice with breast tumor: preliminary results

Figures & data

Figure 1 HSC analysis.

Notes: Samples of MNCs containing HSCs CD34 and CD45 populations were analyzed by flow cytometry. The MNC population was gated in R1 based on FSC and SSC (A), and the expression of CD34 (B) and CD34 versus CD45 (C) was analyzed. These HSCs could form CFUs in culture (D) (scale bar 100 μm).
Abbreviations: HSCs, hematopoietic stem cells; MNCs, mononuclear cells; FSC, forward scatter; SSC, side scatter; CFUs, colony-forming units; FITC, fluorescein isothiocyanate; APC, antigen-presenting cell.

Figure 2 Humanized mice after 28 days of human HSC transplantation.

Notes: Existence of human CD45 leukocytes in murine peripheral blood (A and B). The homing of human HSCs (CD45⁺CD34⁺) into mouse bone marrow (C and D). Existence of subtype population of human leukocytes in murine blood included CD8 cells (E), CD4 cells (F), CD3 cells (G), CD19 cells (H), CD45 cells (I), and CD56 cells (J).
Abbreviations: HSCs, hematopoietic stem cells; FSC, forward scatter; SSC, side scatter; APC, antigen-presenting cell.

Figure 3 Breast tumor-bearing humanized mice.

Notes: Humanized mouse with breast tumor under white light (A) and under GFP excitation (B). Humanized mouse with metastasis to bone (C). The Kaplan–Meier test of survival percentage of humanized mice bearing tumors showed that humanized mice bearing tumors had significantly decreased survival percentage compared to humanized mice (D) and were confirmed by Log-rank test and Gehan-Breslow-Wilcoxon test (E).
Abbreviations: FITC, fluorescein isothiocyanate; sig, significantly; GFPs, green fluorescent proteins.

Figure 4 UCB-derived DCs.

Notes: Expression of specific markers of DCs before being induced with GM-CSF and IL-4 (A–D) and after being induced with GM-CSF and IL-4 and BCSC antigen (E–H). MNCs included with GM-CSF and IL-4 and BCSC antigen strongly expressed the DC markers such as CD40, CD80, CD86, and HLA-DR. These cells also successfully stimulated T-lymphocytes (I) and produced IL-12 (J). Error bars were calculated as standard errors for triplicate.
Abbreviations: UCB, umbilical cord blood; DCs, dendritic cells; GM-CSF, granulocyte macrophage colony-stimulating factor; IL, interleukin; BCSC, breast cancer stem cell; MNCs, mononuclear cells; HLA, human leukocyte antigen; iDC, immature dendritic cell; mDC, mature dendritic cell.

Figure 5 Treatment efficacy was analyzed by Kaplan–Meier and Log-rank test. Kaplan–Meier survival percentage of mice in control (black line) and treatment (red line) groups (A); and confirmed by Log-rank test and Gehan-Breslow-Wilcoxon test (B).

Notes: Humanized mice bearing tumors treated with mature DCs had significantly increased survival percentage P<0.05. **indicate significant differences.
Abbreviation: DC, dendritic cell.