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Myeloid expression of angiotensin-converting enzyme facilitates myeloid maturation and inhibits the development of myeloid-derived suppressor cells.

Shen XZ, Okwan-Duodu D, Blackwell WL, Ong FS, Janjulia T, Bernstein EA, Fuchs S, Alkan S, Bernstein KE - Lab. Invest. (2014)

Bottom Line: Here, we show that ACE expression correlates with myeloid maturation in vitro.Forced ACE overexpression in monocytic cells reduces the generation of MDSCs.Thus, manipulating myeloid ACE activity can interfere with MDSC development and the maturation of myeloid cells.

View Article: PubMed Central - PubMed

Affiliation: 1] Division of Immunology, Department of Biomedical Science, Cedars-Sinai Medical Center, Los Angeles, CA, USA [2] Department of Pathology and Laboratory Medicine, Cedars-Sinai Medical Center, Los Angeles, CA, USA.

ABSTRACT
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells which accumulate in cancer, infection and chronic inflammation. These cells suppress T-cell function and the immune response. Angiotensin-converting enzyme (ACE) is a peptidase that is now known to regulate aspects of myelopoiesis. Here, we show that ACE expression correlates with myeloid maturation in vitro. Forced ACE overexpression in monocytic cells reduces the generation of MDSCs. In vivo, mice with a genetic change resulting in myeloid cell ACE overexpression have reduced numbers of blood and splenic MDSCs in a tumor model and in a model of chronic inflammation induced by complete Freund's adjuvant. In contrast, ACE- mice produce large numbers of MDSCs during chronic inflammation. Macrophages from mice with myeloid ACE overexpressing are more pro-inflammatory and have more tumor-killing activity than cells from wild-type mice. Thus, manipulating myeloid ACE activity can interfere with MDSC development and the maturation of myeloid cells.

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ACE over-expression induces myeloid maturation. a. The top portion of this panel shows the number of Ly6C+Ly6G+ and Ly6C−Ly6G− cells present in the cultured BM from WT and ACE 10/10 mice. ** P<0.01. The bottom portion of this panel is a typical flow cytometric analysis of the WT and ACE 10/10 cells. b. The expression levels of MHC class II and CD86 were measured on the surface of WT and ACE 10/10 BM-derived Ly6C+Ly6G+ cells. c. The non-adherent cells derived from WT and ACE 10/10 BM were co-incubated with stimulated CFSE-labeled T cells. Cytometric analysis after 3 days shows less suppression of T cell proliferation in those cells co-cultured with the ACE 10/10 cells. d. WT BM was transfected with vectors expressing either ACE or a catalytic domain mutated ACE (mACE). The surface expression of MHC class II, CD86 and IL-4Rα was measured after a 4 day culture. e. The non-adherent cells collected from ACE or mACE transfected BM cells were co-cultured with stimulated CFSE-labeled T cells. The T cell proliferation profiles are presented and show less suppression of T cell proliferation by those cells transfected with the catalytically active ACE construct. For all the experiments, the representative flow cytometry figures are typical of at least 3 independent experiments.
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Figure 2: ACE over-expression induces myeloid maturation. a. The top portion of this panel shows the number of Ly6C+Ly6G+ and Ly6C−Ly6G− cells present in the cultured BM from WT and ACE 10/10 mice. ** P<0.01. The bottom portion of this panel is a typical flow cytometric analysis of the WT and ACE 10/10 cells. b. The expression levels of MHC class II and CD86 were measured on the surface of WT and ACE 10/10 BM-derived Ly6C+Ly6G+ cells. c. The non-adherent cells derived from WT and ACE 10/10 BM were co-incubated with stimulated CFSE-labeled T cells. Cytometric analysis after 3 days shows less suppression of T cell proliferation in those cells co-cultured with the ACE 10/10 cells. d. WT BM was transfected with vectors expressing either ACE or a catalytic domain mutated ACE (mACE). The surface expression of MHC class II, CD86 and IL-4Rα was measured after a 4 day culture. e. The non-adherent cells collected from ACE or mACE transfected BM cells were co-cultured with stimulated CFSE-labeled T cells. The T cell proliferation profiles are presented and show less suppression of T cell proliferation by those cells transfected with the catalytically active ACE construct. For all the experiments, the representative flow cytometry figures are typical of at least 3 independent experiments.

Mentions: To investigate the role of ACE in MDSC generation, we used BM from a mouse line termed ACE 10/10 which over-expresses ACE in myeloid cells.17 The culture of ACE 10/10 BM yielded comparable numbers of non-adherent cells as wild-type (WT) BM culture. However, in the non-adherent cell population, the ACE 10/10 cultured cells consistently yielded more of the relatively mature Ly6C−Ly6G− cells and fewer of the immunosuppressive Ly6C+Ly6G+ cells (Figure 2a). Further, detailed analysis of only the Ly6C+Ly6G+ cell population indicated that those cells derived from ACE 10/10 BM expressed higher levels of MHC class II (48% increase in mean fluorescent intensity (MFI)) and CD86 (31% increase in MFI) than equivalent cells derived from WT BM (Figure 2b). Thus, these data suggest that non-adherent cells derived from ACE 10/10 BM, a mouse line over-expressing ACE in myelomonocytic cells, appears somewhat more differentiated and less able to suppress T cell proliferation than an equivalent population from WT BM (Figure 2c).


Myeloid expression of angiotensin-converting enzyme facilitates myeloid maturation and inhibits the development of myeloid-derived suppressor cells.

Shen XZ, Okwan-Duodu D, Blackwell WL, Ong FS, Janjulia T, Bernstein EA, Fuchs S, Alkan S, Bernstein KE - Lab. Invest. (2014)

ACE over-expression induces myeloid maturation. a. The top portion of this panel shows the number of Ly6C+Ly6G+ and Ly6C−Ly6G− cells present in the cultured BM from WT and ACE 10/10 mice. ** P<0.01. The bottom portion of this panel is a typical flow cytometric analysis of the WT and ACE 10/10 cells. b. The expression levels of MHC class II and CD86 were measured on the surface of WT and ACE 10/10 BM-derived Ly6C+Ly6G+ cells. c. The non-adherent cells derived from WT and ACE 10/10 BM were co-incubated with stimulated CFSE-labeled T cells. Cytometric analysis after 3 days shows less suppression of T cell proliferation in those cells co-cultured with the ACE 10/10 cells. d. WT BM was transfected with vectors expressing either ACE or a catalytic domain mutated ACE (mACE). The surface expression of MHC class II, CD86 and IL-4Rα was measured after a 4 day culture. e. The non-adherent cells collected from ACE or mACE transfected BM cells were co-cultured with stimulated CFSE-labeled T cells. The T cell proliferation profiles are presented and show less suppression of T cell proliferation by those cells transfected with the catalytically active ACE construct. For all the experiments, the representative flow cytometry figures are typical of at least 3 independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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Figure 2: ACE over-expression induces myeloid maturation. a. The top portion of this panel shows the number of Ly6C+Ly6G+ and Ly6C−Ly6G− cells present in the cultured BM from WT and ACE 10/10 mice. ** P<0.01. The bottom portion of this panel is a typical flow cytometric analysis of the WT and ACE 10/10 cells. b. The expression levels of MHC class II and CD86 were measured on the surface of WT and ACE 10/10 BM-derived Ly6C+Ly6G+ cells. c. The non-adherent cells derived from WT and ACE 10/10 BM were co-incubated with stimulated CFSE-labeled T cells. Cytometric analysis after 3 days shows less suppression of T cell proliferation in those cells co-cultured with the ACE 10/10 cells. d. WT BM was transfected with vectors expressing either ACE or a catalytic domain mutated ACE (mACE). The surface expression of MHC class II, CD86 and IL-4Rα was measured after a 4 day culture. e. The non-adherent cells collected from ACE or mACE transfected BM cells were co-cultured with stimulated CFSE-labeled T cells. The T cell proliferation profiles are presented and show less suppression of T cell proliferation by those cells transfected with the catalytically active ACE construct. For all the experiments, the representative flow cytometry figures are typical of at least 3 independent experiments.
Mentions: To investigate the role of ACE in MDSC generation, we used BM from a mouse line termed ACE 10/10 which over-expresses ACE in myeloid cells.17 The culture of ACE 10/10 BM yielded comparable numbers of non-adherent cells as wild-type (WT) BM culture. However, in the non-adherent cell population, the ACE 10/10 cultured cells consistently yielded more of the relatively mature Ly6C−Ly6G− cells and fewer of the immunosuppressive Ly6C+Ly6G+ cells (Figure 2a). Further, detailed analysis of only the Ly6C+Ly6G+ cell population indicated that those cells derived from ACE 10/10 BM expressed higher levels of MHC class II (48% increase in mean fluorescent intensity (MFI)) and CD86 (31% increase in MFI) than equivalent cells derived from WT BM (Figure 2b). Thus, these data suggest that non-adherent cells derived from ACE 10/10 BM, a mouse line over-expressing ACE in myelomonocytic cells, appears somewhat more differentiated and less able to suppress T cell proliferation than an equivalent population from WT BM (Figure 2c).

Bottom Line: Here, we show that ACE expression correlates with myeloid maturation in vitro.Forced ACE overexpression in monocytic cells reduces the generation of MDSCs.Thus, manipulating myeloid ACE activity can interfere with MDSC development and the maturation of myeloid cells.

View Article: PubMed Central - PubMed

Affiliation: 1] Division of Immunology, Department of Biomedical Science, Cedars-Sinai Medical Center, Los Angeles, CA, USA [2] Department of Pathology and Laboratory Medicine, Cedars-Sinai Medical Center, Los Angeles, CA, USA.

ABSTRACT
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells which accumulate in cancer, infection and chronic inflammation. These cells suppress T-cell function and the immune response. Angiotensin-converting enzyme (ACE) is a peptidase that is now known to regulate aspects of myelopoiesis. Here, we show that ACE expression correlates with myeloid maturation in vitro. Forced ACE overexpression in monocytic cells reduces the generation of MDSCs. In vivo, mice with a genetic change resulting in myeloid cell ACE overexpression have reduced numbers of blood and splenic MDSCs in a tumor model and in a model of chronic inflammation induced by complete Freund's adjuvant. In contrast, ACE- mice produce large numbers of MDSCs during chronic inflammation. Macrophages from mice with myeloid ACE overexpressing are more pro-inflammatory and have more tumor-killing activity than cells from wild-type mice. Thus, manipulating myeloid ACE activity can interfere with MDSC development and the maturation of myeloid cells.

Show MeSH
Related in: MedlinePlus