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Inhibition of breast cancer resistance protein (ABCG2) in human myeloid dendritic cells induces potent tolerogenic functions during LPS stimulation.

Jin JO, Zhang W, Wong KW, Kwak M, van Driel IR, Yu Q - PLoS ONE (2014)

Bottom Line: Moreover, inhibition of ABCG2 in monocyte-derived DCs (MDDCs) abrogated the up-regulation of co-stimulatory molecules and production of pro-inflammatory cytokines in these cells in response to LPS.Furthermore, CD1c+ mDCs stimulated with LPS plus Ko143 inhibited the proliferation of allogeneic and superantigen-specific syngenic CD4+ T cells and promoted expansion of CD25+FOXP3+ regulatory T (Treg) cells in an IL-10-dependent fashion.These tolerogenic effects of ABCG2 inhibition could be abolished by ERK inhibition.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Public Health Clinical Center, Shanghai Medical College, Fudan University, Shanghai, China.

ABSTRACT
Breast cancer resistance protein (ABCG2), a member of the ATP-binding cassette transporters has been identified as a major determinant of multidrug resistance (MDR) in cancer cells, but ABC transporter inhibition has limited therapeutic value in vivo. In this research, we demonstrated that inhibition of efflux transporters ABCG2 induced the generation of tolerogenic DCs from human peripheral blood myeloid DCs (mDCs). ABCG2 expression was present in mDCs and was further increased by LPS stimulation. Treatment of CD1c+ mDCs with an ABCG2 inhibitor, Ko143, during LPS stimulation caused increased production of IL-10 and decreased production of pro-inflammatory cytokines and decreased expression of CD83 and CD86. Moreover, inhibition of ABCG2 in monocyte-derived DCs (MDDCs) abrogated the up-regulation of co-stimulatory molecules and production of pro-inflammatory cytokines in these cells in response to LPS. Furthermore, CD1c+ mDCs stimulated with LPS plus Ko143 inhibited the proliferation of allogeneic and superantigen-specific syngenic CD4+ T cells and promoted expansion of CD25+FOXP3+ regulatory T (Treg) cells in an IL-10-dependent fashion. These tolerogenic effects of ABCG2 inhibition could be abolished by ERK inhibition. Thus, we demonstrated that inhibition of ABCG2 in LPS-stimulated mDCs can potently induce tolerogenic potentials in these cells, providing crucial new information that could lead to development of better strategies to combat MDR cancer.

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Generation of Treg cells induced by LPS plus Ko143-treated mDCs is dependent on IL-10.CD1c+ mDCs and CD4 T cells were co-cultured as described in Figure 4 in the presence of anti-IL-10 or control IgG Abs. (A) Expression of CD25 and CFSE dilution were analyzed in CD4+ T cells. (B) CD4, CD25 and FOXP3 expression was analyzed by flow cytometry. (C) IFN-γ (left panel) and IL-10 (right panel) concentrations in the culture medium were measured by ELISA. Data are representative or the average of analyses of 3 samples from 3 donors.
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pone-0104753-g005: Generation of Treg cells induced by LPS plus Ko143-treated mDCs is dependent on IL-10.CD1c+ mDCs and CD4 T cells were co-cultured as described in Figure 4 in the presence of anti-IL-10 or control IgG Abs. (A) Expression of CD25 and CFSE dilution were analyzed in CD4+ T cells. (B) CD4, CD25 and FOXP3 expression was analyzed by flow cytometry. (C) IFN-γ (left panel) and IL-10 (right panel) concentrations in the culture medium were measured by ELISA. Data are representative or the average of analyses of 3 samples from 3 donors.

Mentions: Previous studies have reported an essential role for IL-10 in the generation of Treg cells induced by tolerogenic DCs [24], [25]. To determine whether the promoting effect of LPS and Ko143-treated mDCs on Treg generation is dependent on IL-10, we prepared mDCs as described in Figure 4 and co-cultured them with allogeneic naïve CD4 T cells in the presence of a neutralizing anti-IL-10 Abs or its control IgG for 4 days. The results showed that the promoting effect of Ko143- and LPS-treated mDCs on CD25+ T cells was almost completely blocked by the anti-IL-10 Abs (Figure 5A). In contrast, anti-IL-10 Abs treatment caused a small increase in CD4+CD25− T cell proliferation compare to control IgG treatment. Moreover, intranuclear staining showed that anti-IL-10 Abs markedly reduced FOXP3+ Treg cell generation induced by LPS plus Ko143-treated mDCs (Figure 6B). Furthermore, anti-IL-10 Abs treatment partially reversed the inhibition of T cell IFN-γ production and promotion of IL-10 production LPS plus Ko143-treated mDCs (Figure 5C). Hence, these data demonstrate that Treg cell generation induced by LPS plus Ko143-stimulated mDCs is dependent on IL-10.


Inhibition of breast cancer resistance protein (ABCG2) in human myeloid dendritic cells induces potent tolerogenic functions during LPS stimulation.

Jin JO, Zhang W, Wong KW, Kwak M, van Driel IR, Yu Q - PLoS ONE (2014)

Generation of Treg cells induced by LPS plus Ko143-treated mDCs is dependent on IL-10.CD1c+ mDCs and CD4 T cells were co-cultured as described in Figure 4 in the presence of anti-IL-10 or control IgG Abs. (A) Expression of CD25 and CFSE dilution were analyzed in CD4+ T cells. (B) CD4, CD25 and FOXP3 expression was analyzed by flow cytometry. (C) IFN-γ (left panel) and IL-10 (right panel) concentrations in the culture medium were measured by ELISA. Data are representative or the average of analyses of 3 samples from 3 donors.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128747&req=5

pone-0104753-g005: Generation of Treg cells induced by LPS plus Ko143-treated mDCs is dependent on IL-10.CD1c+ mDCs and CD4 T cells were co-cultured as described in Figure 4 in the presence of anti-IL-10 or control IgG Abs. (A) Expression of CD25 and CFSE dilution were analyzed in CD4+ T cells. (B) CD4, CD25 and FOXP3 expression was analyzed by flow cytometry. (C) IFN-γ (left panel) and IL-10 (right panel) concentrations in the culture medium were measured by ELISA. Data are representative or the average of analyses of 3 samples from 3 donors.
Mentions: Previous studies have reported an essential role for IL-10 in the generation of Treg cells induced by tolerogenic DCs [24], [25]. To determine whether the promoting effect of LPS and Ko143-treated mDCs on Treg generation is dependent on IL-10, we prepared mDCs as described in Figure 4 and co-cultured them with allogeneic naïve CD4 T cells in the presence of a neutralizing anti-IL-10 Abs or its control IgG for 4 days. The results showed that the promoting effect of Ko143- and LPS-treated mDCs on CD25+ T cells was almost completely blocked by the anti-IL-10 Abs (Figure 5A). In contrast, anti-IL-10 Abs treatment caused a small increase in CD4+CD25− T cell proliferation compare to control IgG treatment. Moreover, intranuclear staining showed that anti-IL-10 Abs markedly reduced FOXP3+ Treg cell generation induced by LPS plus Ko143-treated mDCs (Figure 6B). Furthermore, anti-IL-10 Abs treatment partially reversed the inhibition of T cell IFN-γ production and promotion of IL-10 production LPS plus Ko143-treated mDCs (Figure 5C). Hence, these data demonstrate that Treg cell generation induced by LPS plus Ko143-stimulated mDCs is dependent on IL-10.

Bottom Line: Moreover, inhibition of ABCG2 in monocyte-derived DCs (MDDCs) abrogated the up-regulation of co-stimulatory molecules and production of pro-inflammatory cytokines in these cells in response to LPS.Furthermore, CD1c+ mDCs stimulated with LPS plus Ko143 inhibited the proliferation of allogeneic and superantigen-specific syngenic CD4+ T cells and promoted expansion of CD25+FOXP3+ regulatory T (Treg) cells in an IL-10-dependent fashion.These tolerogenic effects of ABCG2 inhibition could be abolished by ERK inhibition.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Public Health Clinical Center, Shanghai Medical College, Fudan University, Shanghai, China.

ABSTRACT
Breast cancer resistance protein (ABCG2), a member of the ATP-binding cassette transporters has been identified as a major determinant of multidrug resistance (MDR) in cancer cells, but ABC transporter inhibition has limited therapeutic value in vivo. In this research, we demonstrated that inhibition of efflux transporters ABCG2 induced the generation of tolerogenic DCs from human peripheral blood myeloid DCs (mDCs). ABCG2 expression was present in mDCs and was further increased by LPS stimulation. Treatment of CD1c+ mDCs with an ABCG2 inhibitor, Ko143, during LPS stimulation caused increased production of IL-10 and decreased production of pro-inflammatory cytokines and decreased expression of CD83 and CD86. Moreover, inhibition of ABCG2 in monocyte-derived DCs (MDDCs) abrogated the up-regulation of co-stimulatory molecules and production of pro-inflammatory cytokines in these cells in response to LPS. Furthermore, CD1c+ mDCs stimulated with LPS plus Ko143 inhibited the proliferation of allogeneic and superantigen-specific syngenic CD4+ T cells and promoted expansion of CD25+FOXP3+ regulatory T (Treg) cells in an IL-10-dependent fashion. These tolerogenic effects of ABCG2 inhibition could be abolished by ERK inhibition. Thus, we demonstrated that inhibition of ABCG2 in LPS-stimulated mDCs can potently induce tolerogenic potentials in these cells, providing crucial new information that could lead to development of better strategies to combat MDR cancer.

Show MeSH
Related in: MedlinePlus