Generation of mouse pluripotent stem cell-derived proliferating myeloid cells as an unlimited source of functional antigen-presenting cells

Rong Zhang, Tian Yi Liu, Satoru Senju, Miwa Haruta, Narumi Hirosawa, Motoharu Suzuki, Minako Tatsumi, Norihiro Ueda, Hiroyuki Maki, Ryusuke Nakatsuka, Yoshikazu Matsuoka, Yutaka Sasaki, Shinobu Tsuzuki, Hayao Nakanishi, Ryoko Araki, Masumi Abe, Yoshiki Akatsuka, Yasushi Sakamoto, Yoshiaki Sonoda, Yasuharu Nishimura & 2 others Kiyotaka Kuzushima, Yasushi Uemura

Research output: Contribution to journalArticle

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Abstract

The use of dendritic cells (DC) to prime tumor-associated antigen-specific T-cell responses provides a promising approach to cancer immunotherapy. Embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) can differentiate into functional DCs, thus providing an unlimited source of DCs. However, thepreviously established methods of generating practical volumes of DCs from pluripotent stem cells (PSC) require a large number of PSCs at the start of the differentiation culture. In this study, we generated mouse proliferating myeloid cells (pMC) as a source of antigen-presenting cells (APC) using lentivirus-mediated transduction of the c-Myc gene into mouse PSC-derived myeloid cells. The pMCs could propagate almost indefinitely in a cytokine-dependent manner, while retaining their potential to differentiate into functional APCs. After treatment with IL4 plus GM-CSF, the pMCs showed impaired proliferation and differentiated into immature DC-like cells (pMC-DC) expressing low levels of major histocompatibility complex (MHC)-I, MHC-II, CD40, CD80, and CD86. In addition, exposure to maturation stimuli induced the production of TNFa and IL12p70, and enhanced the expression of MHC-II, CD40, and CD86, which is thus suggestive of typical DC maturation. Similar to bone marrow-derived DCs, they stimulated a primary mixed lymphocyte reaction. Furthermore, the in vivo transfer of pMC-DCs pulsed with H-2Kb-restricted OVA257-264 peptide primed OVA-specific cytotoxic T cells and elicited protection in mice against challenge with OVA-expressing melanoma. Overall, myeloid cells exhibiting cytokine-dependent proliferation and DC-like differentiation may be used to address issues associated with the preparation of DCs.

Original languageEnglish
Pages (from-to)668-677
Number of pages10
JournalCancer Immunology Research
Volume3
Issue number6
DOIs
Publication statusPublished - 01-06-2015

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Pluripotent Stem Cells
Antigen-Presenting Cells
Myeloid Cells
Dendritic Cells
Major Histocompatibility Complex
Cytokines
T-Lymphocytes
Induced Pluripotent Stem Cells
Lentivirus
Mixed Lymphocyte Culture Test
myc Genes
Cytoprotection
Neoplasm Antigens
Granulocyte-Macrophage Colony-Stimulating Factor
Embryonic Stem Cells
Interleukin-4
Immunotherapy
Cell Differentiation
Melanoma
Bone Marrow

All Science Journal Classification (ASJC) codes

  • Immunology
  • Cancer Research

Cite this

Zhang, Rong ; Liu, Tian Yi ; Senju, Satoru ; Haruta, Miwa ; Hirosawa, Narumi ; Suzuki, Motoharu ; Tatsumi, Minako ; Ueda, Norihiro ; Maki, Hiroyuki ; Nakatsuka, Ryusuke ; Matsuoka, Yoshikazu ; Sasaki, Yutaka ; Tsuzuki, Shinobu ; Nakanishi, Hayao ; Araki, Ryoko ; Abe, Masumi ; Akatsuka, Yoshiki ; Sakamoto, Yasushi ; Sonoda, Yoshiaki ; Nishimura, Yasuharu ; Kuzushima, Kiyotaka ; Uemura, Yasushi. / Generation of mouse pluripotent stem cell-derived proliferating myeloid cells as an unlimited source of functional antigen-presenting cells. In: Cancer Immunology Research. 2015 ; Vol. 3, No. 6. pp. 668-677.
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abstract = "The use of dendritic cells (DC) to prime tumor-associated antigen-specific T-cell responses provides a promising approach to cancer immunotherapy. Embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) can differentiate into functional DCs, thus providing an unlimited source of DCs. However, thepreviously established methods of generating practical volumes of DCs from pluripotent stem cells (PSC) require a large number of PSCs at the start of the differentiation culture. In this study, we generated mouse proliferating myeloid cells (pMC) as a source of antigen-presenting cells (APC) using lentivirus-mediated transduction of the c-Myc gene into mouse PSC-derived myeloid cells. The pMCs could propagate almost indefinitely in a cytokine-dependent manner, while retaining their potential to differentiate into functional APCs. After treatment with IL4 plus GM-CSF, the pMCs showed impaired proliferation and differentiated into immature DC-like cells (pMC-DC) expressing low levels of major histocompatibility complex (MHC)-I, MHC-II, CD40, CD80, and CD86. In addition, exposure to maturation stimuli induced the production of TNFa and IL12p70, and enhanced the expression of MHC-II, CD40, and CD86, which is thus suggestive of typical DC maturation. Similar to bone marrow-derived DCs, they stimulated a primary mixed lymphocyte reaction. Furthermore, the in vivo transfer of pMC-DCs pulsed with H-2Kb-restricted OVA257-264 peptide primed OVA-specific cytotoxic T cells and elicited protection in mice against challenge with OVA-expressing melanoma. Overall, myeloid cells exhibiting cytokine-dependent proliferation and DC-like differentiation may be used to address issues associated with the preparation of DCs.",
author = "Rong Zhang and Liu, {Tian Yi} and Satoru Senju and Miwa Haruta and Narumi Hirosawa and Motoharu Suzuki and Minako Tatsumi and Norihiro Ueda and Hiroyuki Maki and Ryusuke Nakatsuka and Yoshikazu Matsuoka and Yutaka Sasaki and Shinobu Tsuzuki and Hayao Nakanishi and Ryoko Araki and Masumi Abe and Yoshiki Akatsuka and Yasushi Sakamoto and Yoshiaki Sonoda and Yasuharu Nishimura and Kiyotaka Kuzushima and Yasushi Uemura",
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Zhang, R, Liu, TY, Senju, S, Haruta, M, Hirosawa, N, Suzuki, M, Tatsumi, M, Ueda, N, Maki, H, Nakatsuka, R, Matsuoka, Y, Sasaki, Y, Tsuzuki, S, Nakanishi, H, Araki, R, Abe, M, Akatsuka, Y, Sakamoto, Y, Sonoda, Y, Nishimura, Y, Kuzushima, K & Uemura, Y 2015, 'Generation of mouse pluripotent stem cell-derived proliferating myeloid cells as an unlimited source of functional antigen-presenting cells', Cancer Immunology Research, vol. 3, no. 6, pp. 668-677. https://doi.org/10.1158/2326-6066.CIR-14-0117

Generation of mouse pluripotent stem cell-derived proliferating myeloid cells as an unlimited source of functional antigen-presenting cells. / Zhang, Rong; Liu, Tian Yi; Senju, Satoru; Haruta, Miwa; Hirosawa, Narumi; Suzuki, Motoharu; Tatsumi, Minako; Ueda, Norihiro; Maki, Hiroyuki; Nakatsuka, Ryusuke; Matsuoka, Yoshikazu; Sasaki, Yutaka; Tsuzuki, Shinobu; Nakanishi, Hayao; Araki, Ryoko; Abe, Masumi; Akatsuka, Yoshiki; Sakamoto, Yasushi; Sonoda, Yoshiaki; Nishimura, Yasuharu; Kuzushima, Kiyotaka; Uemura, Yasushi.

In: Cancer Immunology Research, Vol. 3, No. 6, 01.06.2015, p. 668-677.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Generation of mouse pluripotent stem cell-derived proliferating myeloid cells as an unlimited source of functional antigen-presenting cells

AU - Zhang, Rong

AU - Liu, Tian Yi

AU - Senju, Satoru

AU - Haruta, Miwa

AU - Hirosawa, Narumi

AU - Suzuki, Motoharu

AU - Tatsumi, Minako

AU - Ueda, Norihiro

AU - Maki, Hiroyuki

AU - Nakatsuka, Ryusuke

AU - Matsuoka, Yoshikazu

AU - Sasaki, Yutaka

AU - Tsuzuki, Shinobu

AU - Nakanishi, Hayao

AU - Araki, Ryoko

AU - Abe, Masumi

AU - Akatsuka, Yoshiki

AU - Sakamoto, Yasushi

AU - Sonoda, Yoshiaki

AU - Nishimura, Yasuharu

AU - Kuzushima, Kiyotaka

AU - Uemura, Yasushi

PY - 2015/6/1

Y1 - 2015/6/1

N2 - The use of dendritic cells (DC) to prime tumor-associated antigen-specific T-cell responses provides a promising approach to cancer immunotherapy. Embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) can differentiate into functional DCs, thus providing an unlimited source of DCs. However, thepreviously established methods of generating practical volumes of DCs from pluripotent stem cells (PSC) require a large number of PSCs at the start of the differentiation culture. In this study, we generated mouse proliferating myeloid cells (pMC) as a source of antigen-presenting cells (APC) using lentivirus-mediated transduction of the c-Myc gene into mouse PSC-derived myeloid cells. The pMCs could propagate almost indefinitely in a cytokine-dependent manner, while retaining their potential to differentiate into functional APCs. After treatment with IL4 plus GM-CSF, the pMCs showed impaired proliferation and differentiated into immature DC-like cells (pMC-DC) expressing low levels of major histocompatibility complex (MHC)-I, MHC-II, CD40, CD80, and CD86. In addition, exposure to maturation stimuli induced the production of TNFa and IL12p70, and enhanced the expression of MHC-II, CD40, and CD86, which is thus suggestive of typical DC maturation. Similar to bone marrow-derived DCs, they stimulated a primary mixed lymphocyte reaction. Furthermore, the in vivo transfer of pMC-DCs pulsed with H-2Kb-restricted OVA257-264 peptide primed OVA-specific cytotoxic T cells and elicited protection in mice against challenge with OVA-expressing melanoma. Overall, myeloid cells exhibiting cytokine-dependent proliferation and DC-like differentiation may be used to address issues associated with the preparation of DCs.

AB - The use of dendritic cells (DC) to prime tumor-associated antigen-specific T-cell responses provides a promising approach to cancer immunotherapy. Embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) can differentiate into functional DCs, thus providing an unlimited source of DCs. However, thepreviously established methods of generating practical volumes of DCs from pluripotent stem cells (PSC) require a large number of PSCs at the start of the differentiation culture. In this study, we generated mouse proliferating myeloid cells (pMC) as a source of antigen-presenting cells (APC) using lentivirus-mediated transduction of the c-Myc gene into mouse PSC-derived myeloid cells. The pMCs could propagate almost indefinitely in a cytokine-dependent manner, while retaining their potential to differentiate into functional APCs. After treatment with IL4 plus GM-CSF, the pMCs showed impaired proliferation and differentiated into immature DC-like cells (pMC-DC) expressing low levels of major histocompatibility complex (MHC)-I, MHC-II, CD40, CD80, and CD86. In addition, exposure to maturation stimuli induced the production of TNFa and IL12p70, and enhanced the expression of MHC-II, CD40, and CD86, which is thus suggestive of typical DC maturation. Similar to bone marrow-derived DCs, they stimulated a primary mixed lymphocyte reaction. Furthermore, the in vivo transfer of pMC-DCs pulsed with H-2Kb-restricted OVA257-264 peptide primed OVA-specific cytotoxic T cells and elicited protection in mice against challenge with OVA-expressing melanoma. Overall, myeloid cells exhibiting cytokine-dependent proliferation and DC-like differentiation may be used to address issues associated with the preparation of DCs.

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