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Targeting specificity of dendritic cells on breast cancer stem cells: in vitro and in vivo evaluations.

Nguyen ST, Nguyen HL, Pham VQ, Nguyen GT, Tran CD, Phan NK, Pham PV - Onco Targets Ther (2015)

Bottom Line: The results show that in vitro BCSC-DCs significantly inhibited BCSC proliferation at a DC:CTL ratio of 1:40, while MSC-DCs nonsignificantly decreased BCSC proliferation.In vivo, tumor sizes decreased from 18.8% to 23% in groups treated with BCSC-DCs; in contrast, tumors increased 14% in the control group (RPMI 1640) and 47% in groups treated with MSC-DCs.The results showed that DC therapy could target and be specific to BCSCs.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam.

ABSTRACT
Breast cancer is a leading cause of death in women, and almost all complications are due to chemotherapy resistance. Drug-resistant cells with stem cell phenotypes are thought to cause failure in breast cancer chemotherapy. Dendritic cell (DC) therapy is a potential approach to eradicate these cells. This study evaluates the specificity of DCs for breast cancer stem cells (BCSCs) in vitro and in vivo. BCSCs were enriched by a verapamil-resistant screening method, and reconfirmed by ALDH expression analysis and mammosphere assay. Mesenchymal stem cells (MSCs) were isolated from allogeneic murine bone marrow. DCs were induced from bone marrow-derived monocytes with 20 ng/mL GC-MSF and 20 ng/mL IL-4. Immature DCs were primed with BCSC- or MSC-derived antigens to make two kinds of mature DCs: BCSC-DCs and MSC-DCs, respectively. In vitro ability of BCSC-DCs and MSC-DCs with cytotoxic T lymphocytes (CTLs) to inhibit BCSCs was tested using the xCELLigence technique. In vivo, BCSC-DCs and MSC-DCs were transfused into the peripheral blood of BCSC tumor-bearing mice. The results show that in vitro BCSC-DCs significantly inhibited BCSC proliferation at a DC:CTL ratio of 1:40, while MSC-DCs nonsignificantly decreased BCSC proliferation. In vivo, tumor sizes decreased from 18.8% to 23% in groups treated with BCSC-DCs; in contrast, tumors increased 14% in the control group (RPMI 1640) and 47% in groups treated with MSC-DCs. The results showed that DC therapy could target and be specific to BCSCs. DCs primed with MSCs could trigger tumor growth. These results also indicate that DCs may be a promising therapy for treating drug-resistant cancer cells as well as cancer stem cells.

No MeSH data available.


Related in: MedlinePlus

Dendritic cells were generated from mouse bone marrow.Notes: Dendritic cells exhibited the particular morphology on day 5 in medium supplemented with GM-CSF and IL-4 (40× magnification) (A); on day 4 after induction by antigens (40× magnification) (B). The arrows indicate the dendrites on the DCs. Immune phenotype of DCs were characterized by flow cytometry (C–F). They expressed CD40 (D), CD86 (E), and CD80 (F) but lacked expression of CD14 (C).Abbreviations: DC, dendritic cells; GM-CSF, granulocyte-macrophage colony-stimulating factor.
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f4-ott-8-323: Dendritic cells were generated from mouse bone marrow.Notes: Dendritic cells exhibited the particular morphology on day 5 in medium supplemented with GM-CSF and IL-4 (40× magnification) (A); on day 4 after induction by antigens (40× magnification) (B). The arrows indicate the dendrites on the DCs. Immune phenotype of DCs were characterized by flow cytometry (C–F). They expressed CD40 (D), CD86 (E), and CD80 (F) but lacked expression of CD14 (C).Abbreviations: DC, dendritic cells; GM-CSF, granulocyte-macrophage colony-stimulating factor.

Mentions: Bone marrow-derived mononuclear cells were cultured in RPMI medium supplemented with GM-CSF and IL-4 for 7 days to generate immature DCs. Morphological observation on day 7 showed formation of dendrites that are typical of DCs (Figure 4A, B). On day 7, DCs still adhered to the culture flask bottom. These iDCs (immature dendritic cells) expressed the typical phenotypes of DCs, including the presence of CD80, CD86, and CD40 (Figure 4D–F) and the absence of CD14 (Figure 4C). Mature iDCs were induced by adding antigens into the culture medium with cytokines. After 3 days in the medium supplemented with antigens from both BCSCs and MSCs, DCs detached from the flask bottom and floated in the medium. Morphological observation showed that there are veils around the DCs.


Targeting specificity of dendritic cells on breast cancer stem cells: in vitro and in vivo evaluations.

Nguyen ST, Nguyen HL, Pham VQ, Nguyen GT, Tran CD, Phan NK, Pham PV - Onco Targets Ther (2015)

Dendritic cells were generated from mouse bone marrow.Notes: Dendritic cells exhibited the particular morphology on day 5 in medium supplemented with GM-CSF and IL-4 (40× magnification) (A); on day 4 after induction by antigens (40× magnification) (B). The arrows indicate the dendrites on the DCs. Immune phenotype of DCs were characterized by flow cytometry (C–F). They expressed CD40 (D), CD86 (E), and CD80 (F) but lacked expression of CD14 (C).Abbreviations: DC, dendritic cells; GM-CSF, granulocyte-macrophage colony-stimulating factor.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4321654&req=5

f4-ott-8-323: Dendritic cells were generated from mouse bone marrow.Notes: Dendritic cells exhibited the particular morphology on day 5 in medium supplemented with GM-CSF and IL-4 (40× magnification) (A); on day 4 after induction by antigens (40× magnification) (B). The arrows indicate the dendrites on the DCs. Immune phenotype of DCs were characterized by flow cytometry (C–F). They expressed CD40 (D), CD86 (E), and CD80 (F) but lacked expression of CD14 (C).Abbreviations: DC, dendritic cells; GM-CSF, granulocyte-macrophage colony-stimulating factor.
Mentions: Bone marrow-derived mononuclear cells were cultured in RPMI medium supplemented with GM-CSF and IL-4 for 7 days to generate immature DCs. Morphological observation on day 7 showed formation of dendrites that are typical of DCs (Figure 4A, B). On day 7, DCs still adhered to the culture flask bottom. These iDCs (immature dendritic cells) expressed the typical phenotypes of DCs, including the presence of CD80, CD86, and CD40 (Figure 4D–F) and the absence of CD14 (Figure 4C). Mature iDCs were induced by adding antigens into the culture medium with cytokines. After 3 days in the medium supplemented with antigens from both BCSCs and MSCs, DCs detached from the flask bottom and floated in the medium. Morphological observation showed that there are veils around the DCs.

Bottom Line: The results show that in vitro BCSC-DCs significantly inhibited BCSC proliferation at a DC:CTL ratio of 1:40, while MSC-DCs nonsignificantly decreased BCSC proliferation.In vivo, tumor sizes decreased from 18.8% to 23% in groups treated with BCSC-DCs; in contrast, tumors increased 14% in the control group (RPMI 1640) and 47% in groups treated with MSC-DCs.The results showed that DC therapy could target and be specific to BCSCs.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam.

ABSTRACT
Breast cancer is a leading cause of death in women, and almost all complications are due to chemotherapy resistance. Drug-resistant cells with stem cell phenotypes are thought to cause failure in breast cancer chemotherapy. Dendritic cell (DC) therapy is a potential approach to eradicate these cells. This study evaluates the specificity of DCs for breast cancer stem cells (BCSCs) in vitro and in vivo. BCSCs were enriched by a verapamil-resistant screening method, and reconfirmed by ALDH expression analysis and mammosphere assay. Mesenchymal stem cells (MSCs) were isolated from allogeneic murine bone marrow. DCs were induced from bone marrow-derived monocytes with 20 ng/mL GC-MSF and 20 ng/mL IL-4. Immature DCs were primed with BCSC- or MSC-derived antigens to make two kinds of mature DCs: BCSC-DCs and MSC-DCs, respectively. In vitro ability of BCSC-DCs and MSC-DCs with cytotoxic T lymphocytes (CTLs) to inhibit BCSCs was tested using the xCELLigence technique. In vivo, BCSC-DCs and MSC-DCs were transfused into the peripheral blood of BCSC tumor-bearing mice. The results show that in vitro BCSC-DCs significantly inhibited BCSC proliferation at a DC:CTL ratio of 1:40, while MSC-DCs nonsignificantly decreased BCSC proliferation. In vivo, tumor sizes decreased from 18.8% to 23% in groups treated with BCSC-DCs; in contrast, tumors increased 14% in the control group (RPMI 1640) and 47% in groups treated with MSC-DCs. The results showed that DC therapy could target and be specific to BCSCs. DCs primed with MSCs could trigger tumor growth. These results also indicate that DCs may be a promising therapy for treating drug-resistant cancer cells as well as cancer stem cells.

No MeSH data available.


Related in: MedlinePlus