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Online ISSN : 2424-0664
Print ISSN : 0916-6920
ISSN-L : 2424-0664
27 巻 , 1 号
選択された号の論文の7件中1~7を表示しています
原著
  • 坂本 淳, 野間 芳弘
    2017 年 27 巻 1 号 p. 1-6
    発行日: 2017/05/25
    公開日: 2017/05/31
    ジャーナル フリー

    The detection and quantitation of glycosylphosphatidylinositol (GPI) anchor type membrane protein by flow cytometry (FCM) is defined in the diagnosis of paroxymal nocturnal hemoglobinuria (PNH) and is performed as insurance reimbursed test in Japan. However, the sensitivity of this test is 1% and does not meet the requirements for detecting trace amount of PNH clone inbone marrow failure. Therefore, we have performed preliminary study on highly sensitive method for detecting PNH clone that is in compliance with the International Clinical Cytometry Society (ICCS) guideline.

    Preliminary results indicated that the PNH clone ratio (RBC, granulocyte and monocyte) for healthy individual was less than 0.01%. This result is in compliance with the sensitivity specified in ICCS guideline which is 0.01%. Furthermore, the validity of the results were backed by testing PNH series survey samples from Collage of American Pathologists (CAP).However, confirmation by re-testing was recommended for samples around 0.01% due to low reproducibility.

    With the results obtained from this preliminary study, this test was modified to standard operational testing format. The utility of this new testing method was shown to be useful using clinical samples.

総説
  • 西 将康, 山下 公大, 長谷川 寛, 田中 智子, 有本 聡, 山本 将士, 金治 新悟, 松田 佳子, 押切 太郎, 角 泰雄, 中村 ...
    2017 年 27 巻 1 号 p. 7-12
    発行日: 2017/05/25
    公開日: 2017/05/31
    ジャーナル フリー

    Recently, cancer immunotherapy is noticed as a new therapy of cancer, in addition to surgery, chemotherapy, and radiotherapy. We have been focused on alpha-galactosylceramide (αGalCer) loaded cell treatment, that can activate invariant NKT cell, and consequently activate both innate and acquired immunity by intravenous injection. Especially, we focus on allogeneic dendritic cells pulsed with αGalCer (DCG ) treatment because of its therapeutic efficacy. Our research clarified allogeneic DCG treatment prolonged survival similar to syngeneic DCG treatment. And also we revealed allogeneic DCG treatment induced similar antitumor effect regarding NK and NKT cell activity compared to syngeneic DCG treatment. As a result, our data shows allogeneic DCG treatment could be substituted for syngeneic DCG treatment. Furthermore, our results also show that the therapeutic value of allogeneic DCG is significantly better than syngeneic DCG in a kind of tumor cell lines. According to an examination of cytotoxic activity after treatment, this superiority would be caused by enhancement of antitumor effect with acquired immunity. We are currently working on creating therapeutic models by adopting antigen-specific T cells, and examine this enhancement.

  • 関戸 悠紀, 西村 潤一, 高橋 秀和, 原口 直紹, 畑 泰司, 松田 宙, 水島 恒和, 土岐 祐一郎, 森 正樹
    2017 年 27 巻 1 号 p. 13-17
    発行日: 2017/05/25
    公開日: 2017/05/31
    ジャーナル フリー

    For understanding pathology of inflammatory bowel disease, function analysis of immune cells in the human intestinal tract is essential, but many points remain unclear. In recent years we have developed a method to isolate immunocompetent cells from the human intestinal tract. The obtained HLA-DR positive cells could be classified into 4 groups by CD14, CD11c, CD163. The cell population of CD14+CD163low produced inflammatory cytokines such as IL-6, IL23p19, TNF, and the ability to induce Th17 cells was confirmed. CD14+CD163low cells were found in not only inflamed, but also non-inflamed parts of the Crohn’s disease intestinal tract, and the expression of inflammatory cytokines was enhanced and the Th17 cell-inducing ability was markedly enhanced as compared with the normal intestinal tract.

    In addition, CD14+CD163highCD160high cells, a subpopulation of CD14+CD163high cells, inhibited CD14+CD163low cell dependent T cell proliferation. Through these cell population analyses, it was suggested that these cell groups could be a target in the treatment of IBD.

  • 上田 舞, 高島 康弘
    2017 年 27 巻 1 号 p. 19-24
    発行日: 2017/05/25
    公開日: 2017/05/31
    ジャーナル フリー

    Since mouse embryonic stem cells (ESCs) were established from pre-implantation epiblasts, mouse genomics has dramatically progressed and life science has been expanded. After the establishment of human ESCs in 1998 and human induced pluripotent stem cells (iPSCs) in 2007, there has been added expectation for regenerative medicine from pluripotent stem cells (PSCs). However, the pluripotency character of PSCs is not as well studied as the differentiation potential. We still do not know why, for example, mouse and human PSCs have distinct morphologies and signaling requirements for the maintenance of pluripotency as well as different transcriptome and epigenome.

    Currently, PSCs are classified into two pluripotent states, naïve and primed. Mouse PSCs represent naïve state, which corresponds to pre-implantation epiblasts and can contribute to chimaeras efficiently, whereas human PSCs represent primed state, which corresponds to post-implantation epiblasts.

    We have recently succeeded in generating human naïve PSCs by transient overexpression of NANOG and KLF2 in human primed PSCs. Our human naïve PSCs and mouse ESCs/iPSCs have similar gene expression pattern. Furthermore, they have DNA hypomethylation and alteration in mitochondrial metabolism that resemble preimplantation epiblasts. Human naïve PSCs may be useful to overcome the problems of heterogeneous populations, variation between cell lines, and the poorer differentiation efficiency of human PSCs compared with mouse PSCs. Additionally, naïve human PSCs open the gate to study human pre-implantation embryogenesis.

    In this review, we have written the history, current topics and future direction of PSCs and human naïve PSCs.

  • 岡田 誠治
    2017 年 27 巻 1 号 p. 25-31
    発行日: 2017/05/25
    公開日: 2017/05/31
    ジャーナル フリー

    Development of novel strains of immunodeficient mice has begun to provide the opportunity to utilize mice model for the study of human specific diseases and human stem cell research. The introduction of NOD/Scid based immunodeficient mice such as NOG (NOD/Scid/IL2Rγnull), NSG (NOD/Scid/IL2Rγnull), and NOJ (NOD/Scid/Jak3null) mice, led to a breakthrough in the ability to engraft hematopoietic stem cells effectively creating human hematopoietic and immune systems in these mice. These humanized mice are now becoming increasing important as pre-clinical model for the study of HIV and other human specific infectious diseases. Recently, patient derived tumor xenograft (PDX) for these highly immunodeficient mice are also in the spotlight, since PDX at low passage have been shown to conserve original tumor characteristics such as heterogeneous histology, clinical biomolecular signature, malignant phenotypes and genotypes, tumor architecture and tumor vasculature. These mice are also applied for regenerative medicine using tissue specific stem cells and iPS. Thus, these human xenograft animal models contribute to study human immunology, stem cell biology, and cancer research, and are getting powerful tool for advancing Precision Medicine. Development of novel strains of immunodeficient mice has begun to provide the opportunity for the study of human specific diseases and human stem cell research. The intrimmunodeficient mice such as NOG (NOD/Scid/IL2Rγnull), NSG (NOD/Scid/IL2Rγnull), mice, led to a breakthrough in the ability to engraft hematopoietic stem cells effectively and immune systems in these mice. These humanized mice are now becoming increasing model for the study of HIV and other human specific infectious diseases. Recently, patient derived tumor xenograft (PDX)for these highly immunodeficient mice are also in the spotlight, since PDX at low passage immunodeficient mice such as NOG (NOD/Scid/IL2Rγnull), NSG (NOD/Scid/IL2Rγnull), and NOJ (NOD/Scid/Jak3null) mice, led to a breakthrough in the ability to engraft hematopoietic stem cells effectively creating human hematopoietic and immune systems in these mice. These humanized mice are now becoming increasing important as pre-clinical model for the study of HIV and other human specific infectious diseases. Recently, patient derived tumor xenograft (PDX) for these highly immunodeficient mice are also in the spotlight, since PDX at low passage have been shown to conserve original tumor characteristics such as heterogeneous histology, clinical biomolecular signature, malignant phenotypes and genotypes, tumor architecture and tumor vasculature. These mice are also applied for regenerative medicine using tissue specific stem cells and iPS. Thus, these human xenograft animal models contribute to study human immunology, stem cell biology, and cancer research, and are getting powerful tool for advancing Precision Medicine.

  • 下野 洋平, 渋谷 尚樹, 向山 順子, 西村 建徳, 後藤 典子, 南 博信, 掛地 吉弘, 鈴木 聡
    2017 年 27 巻 1 号 p. 33-39
    発行日: 2017/05/25
    公開日: 2017/05/31
    ジャーナル フリー

    Human cancers, such as breast and colon cancers, contain cancer stem cells (CSCs) which are involved in various aspects of tumor progression, such as recurrence, metastasis, and drug resistance. The fact that metastases can appear years or even decades after the surgical removal of the primary tumor strongly suggests a critical role of metastatic CSCs (mCSCs) in cancer progression.

    miRNAs are short non-coding RNAs that regulate various cellular processes, such as differentiation, proliferation, apoptosis, and stem cell maintenance. We have previously reported that miRNAs, such as miR-200c, miR-183, and miR-142, are differentially expressed between human breast CSCs and other non-tumorigenic cancer cells. Both miR-200c and miR-183 suppress the expression of a self-renewal gene BMI1. miR-142 targets APC and activates the WNT signaling pathway that is implicated in stem cell maintenance.

    Then, we established the PDX models of human breast cancers and compared miRNA expression between CSCs at the primary site (pCSCs) and mCSCs. Although both pCSCs and mCSCs were within a CD44+/CD24-/low population, the expression profile of miRNAs was different between pCSCs and mCSCs. In addition, miRNAs that were downregulated in mCSCs than in pCSCs had potential to suppress the canonical WNT signaling activity, suggesting that the enhancement of WNT signaling activities is important to maintain mCSCs at the metastatic site.

    Because mCSCs in metastatic regions are more resistant to cancer therapy, clarifying the molecular regulation of the human breast mCSCs will further advance our understanding of the roles of human breast CSCs in tumor progression and therapeutic resistance.

実験手技
  • 吉川 倫太郎, 小松 貴義, 宮内 裕美, 陶山 隆史, 宮本 憲一, 松崎 有未
    2017 年 27 巻 1 号 p. 41-45
    発行日: 2017/05/25
    公開日: 2017/05/31
    ジャーナル フリー

    間葉系幹細胞 (Mesenchymal stem cells:MSCs) は,非血球系でプラスチックへの接着性があり,自己複製能と脂肪・骨・軟骨への分化能を有する細胞であると定義されている。MSCsは適切な培養条件に置くことで筋細胞・神経様細胞・肝細胞など,間質以外の細胞に分化することも報告されているため,これらの組織の難治性疾患に対する細胞移植療法のための細胞供給源として期待されている。さらに近年ではMSCsに免疫抑制能があることが示され,さまざまな免疫疾患に対する治療法への応用が検討されている。すでに2015年より本邦でヒトMSCsが急性移植片対宿主病を適応症として製造販売されている。

    MSCsはin vitroで容易に培養可能であることから,通常骨髄細胞を培養皿上で培養後に付着増殖した細胞を回収する方法で分離されている。しかしこの方法で分離された細胞集団には,MSCs以外に前駆細胞・脂肪細胞・マクロファージ・内皮細胞等が混入する可能性がある。また,数週間の培養期間を要するため,分化能や遊走能の喪失および細胞老化などMSCs自体の性質変化を引き起こすという問題点を抱えていた。

    このような問題を解決するために,われわれは骨髄から直接マウスMSCsを分離できるような特異的細胞表面マーカーの同定を試みた。その結果,マウスMSCsはPDGFR−α(platelet-derived growth factor receptor-alpha)とSca−1(stem cell antigen−1)の共陽性分画に含まれることを明らかにした。このMSCs特異的細胞表面マーカーを指標としたフローサイトメトリーによるMSCs分離法を用いることで,培養条件や他の細胞の影響のない均一なMSCsを得ることが可能となった。この方法は,これまで困難であったMSCsの生体内分布や生理機能の解析を可能にするものとして期待される。

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