著者

安田 忠仁 石本 崇胤

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.32, no.1, pp.27-32, 2022-07-13 (Released:2022-07-13)

参考文献数

11

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Recent studies have revealed senescent non-malignant cells in the tumor microenvironment exhibit a secretory profi le under stress conditions; this senescence-associated secretory phenotype (SASP) promotes carcinogenesis and cancer progression. However, the role of senescent non-malignant cells in the metastatic process is not well-understood. We showed that fi broblast population shows p16 expression and SASP factors at high levels in the ascites of gastric cancer patients with peritoneal dissemination by single-cell mass cytometry (CyTOF). Moreover, we present the senolysis strategy in the tumor microenvironment based on the results of an in vivo validation. We identifi ed piperlongumine as the effective senolytic drug for senescent-fi broblasts. These fi ndings offer some notice toward a successful treatment targeting harmful senescent cells and provide the potential for clinical application for the future therapeutic strategy for combining conventional chemotherapy. progression. However, the role of senescent non-malignant cells in the metastatic process is not well-understood. We showed that fi broblast population shows p16 expression and SASP factors at high levels in the ascites of gastric cancer patients with peritoneal dissemination by single-cell mass cytometry (CyTOF). Moreover, we present the senolysis strategy in the tumor microenvironment based on the results of an in vivo validation. We identifi ed piperlongumine as the effective senolytic drug for senescent-fi broblasts. These fi ndings offer some notice toward a successful treatment targeting harmful senescent cells and provide the potential for clinical application for the future therapeutic strategy for combining conventional chemotherapy.

著者

伊藤 量基

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.25, no.1, pp.19-24, 2015-04-25 (Released:2017-05-08)

参考文献数

20

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Dendritic cells (DCs) are the master cells in activating immune responses through enhancing innate immunity by cytokine production and initiating acquired immunity by priming naïve CD4+ T cells. However, DCs play not only a central role in antimicrobial immune response in host defense but also a pathogenic role in the development of the several inflammatory disorders such as allergy and autoimmune diseases. In allergy, epithelial cell-derived thymic stromal lymphopoietin stimulates myeloid DC subset to express OX40L and CCL17 that induce and maintain Th2 cell responses. In systemic lupus erythematosus (SLE), aberrant continuous type I IFN production by plasmacytoid DCs causes vicious spiral of pathogenic autoimmune responses. Thus, myeloid DC and plasmacytoid DC subsets represent key cellular pathogenic cells in allergy and SLE, respectively. Accordingly, control of the dysregulated myeloid DC functions and pDC-derived type I IFNs provides an alternative treatment strategy for allergy and SLE. We focus on specific roles of statins in controlling the myeloid DCs-dependent Th2 pathway and plasmacytoid DC-dependent IFN pathway and state their therapeutic potentials for allergy and SLE.

著者

笹土 隆雄 竹花 佑介 成瀬 清

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.24, no.2, pp.1-7, 2014-10-25 (Released:2017-05-19)

参考文献数

28

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Medaka has been developed as a model animal that can be used to apply both forward and reverse genetic approaches. Large-scale mutagenesis screening for developmental processes, full-length cDNA/Expressed sequence Tag(EST) sequencing and genome sequencing projects have made it possible to use medaka as an effective model for research studies using a forward genetic approach. Currently, it is possible to identify the causal genes in medaka mutants within one to two years. Moreover, the TILLING library and high-throughput screening of mutations by highresolution melting (HRM) facilitate the identification of mutations in a particular gene and can produce medaka with mutations in the particular gene of interest. In addition to this reverse genetics approach, genome editing with the use of engineered nucleases, such as TALEN and CRISPR/CAS9, can be applicable to medaka. Recently, knock-in expression with a GFP cassette at the de novo locus was reported in zebrafish, and this can also be applied to medaka. Therefore, most methods for forward and reverse genetic approaches are equivalent or easier with the medaka model than other animal models such as the mouse and rat.Although constant environments (fixed temperature and day/night cycle) are the general condition for animal experiments, organisms in nature live in fluctuating environments. Thus, studies on phenotype/genotype interactions in fluctuating environments represent the future for the biological and biomedical sciences. In this context, numerous features of medaka, such as their adaptability to variable temperature (4 to 40 ℃), tolerance to high-salinity environments without acclimatization, ability to measure the light/dark cycle and adaptation to seasonal change, can provide important information for analyses of phenotype, genotype and environmental interaction. For these reasons, we believe that medaka, a model animal established in Japan, is a good candidate for experimental animal studies of phenotype and genotype interactions in changing environments.

著者

岡田 誠治

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.27, no.2, pp.51-56, 2017-11-25 (Released:2017-11-25)

参考文献数

22

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Patient-derived xenograft (PDX) models can be created with transplantation of cancerous cells or tissues from patient’s primary tumors into immunodeficient mice. PDX technology leads to the breakthrough with the introduction of novel highly immunodeficient mice such as NOG (NOD/Scid/IL2Rγnull), NSG (NOD/Scid/IL2Rγnull), and NOJ (NOD/Scid/Jak3null) mice. As PDX has been shown to conserve human original tumor characteristics, it is now getting standard methods to evaluate the drug sensitivity, and PDX library is now established in many organization with integrated genomic signature. These PDX data base are getting powerful tool for advancing Precision Cancer Medicine.

著者

佐竹 敦志

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.25, no.2, pp.7-13, 2015-10-25 (Released:2017-05-08)

参考文献数

18

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Strategies to expand regulatory T cells (Treg)s hold therapeutic potential for ameliorating T cell-mediated inflammatory diseases. TCR signaling is partially dispensable for IL-2-induced Treg proliferation, as has been shown recently. In contrast, conventional T cells (Tconv)s require TCR signal activation for their proliferation. Therefore, we can expect that in conjunction with IL-2, pharmacological inhibition of TCR signaling may induce the expansion of Tregs while simultaneously inhibiting Tconv proliferation. We hereby demonstrate that costimulation but not TCR-mediated phospholipase Cγ (PLCγ) activation is required for the IL-2-induced Treg proliferation. Using Cyclosporine A (CSA) to inhibit TCR signaling and rapamycin to suppress costimulatory signaling pathways, we also demonstrate that while both agents attenuated antigen-specific Tconv proliferation, only CSA permitted IL-2-induced Treg expansion. Rapamycin, however, did increase the Treg:Tconv ratio due to its negative effects on Tconv survival. Importantly, CSA synergized with IL-2 in protection against experimental autoimmune encephalomyelitis (EAE). However, CSA abolished whereas rapamycin augmented the beneficial effect of IL-2 in graft-versus-host disease (GVHD). These differences could be potentially explained by the ability of rapamycin to promote inducible Treg generation and to allow TCR-enhanced Treg proliferation, processes that were inhibited by CSA but are important for GVHD protection. Thus, depending on the disease setting, different signaling pathways need to be targeted to increase the Treg:Tconv ratio for treatment of T cellmediated inflammatory disorders.

著者

後藤 典子

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.32, no.2, pp.17-22, 2023-04-17 (Released:2023-04-17)

参考文献数

16

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Breast cancer incidence has increased significantly worldwide, and in Japan, one out of every nine women may experience this disease during their lifetime. Despite recent advances in diagnostic and therapeutic strategies, some patients experience relapse several years after treatment and have a poor prognosis. Therefore, it is essential to develop effective treatment strategies to prevent relapse. Recent evidence suggests that subpopulations of cancer stem cells are highly resistant to chemotherapy and radiotherapy, allowing them to survive after treatment. These cancer stem cells were fi rst discovered in breast cancer in 2003. Scientists around the world have made efforts to elucidate the molecular mechanisms of these cells, and it is now believed that they survive in a “cancer stem cell niche” in the tumor microenvironment, among other cells such as cancer-associated fibroblasts and immune cells, by interacting with multiple cytokines and growth factors. Recent cutting-edge single-cell technologies have allowed for the elucidation of the molecular mechanisms of these therapy-resistant cancer stem cells. These efforts in basic science may pave the way for the development of effective treatment strategies to target these cells and prevent relapse, ultimately leading to a signifi cant improvement in the prognosis of breast cancer patients.

著者

松崎 正晴

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.15, no.1, pp.15-19, 2005-04-01 (Released:2017-07-04)

参考文献数

11

---

Gene silencing is widely used to clarify the gene function. Although RNA interference (RNAi) can expect a handy and large effect for gene silencing, the development of the screening system of a target gene is important. Green fluorescent proteins (GFPs) and Reef Coral fluorescent proteins (RCFPs) are well known that can make stable fluoresce without any co-factors in almost all cells. Both of the function of the target gene and the fluorescent of GFPs/RCFPs are most probably kept, even if a lot of genes are made as a fusion protein with these fluorescent proteins. When GFP or RCFP is connected with C-terminal of a target protein, and if a target protein receives the resolution by RNAi, the fluorescence of GFP or RCFP in the downstream of a target protein cannot be translated to a protein and fluorescence disappears. We developed the high throughput screening system for RNAi. Fluorescence-Activated Cell Sorter (FACS) measurement of the fluorescence of 293 cells after the expression vector of a target gene and RNAi vector were co-transfected to 293 cells, a handy screening of RNAi can be established.

著者

清河 信敬 恩田 恵子 高野 邦彦 藤本 純一郎 真部 淳 康 勝好 小原 明 林 泰秀 花田 良二 土田 昌宏

出版者

日本サイトメトリー学会

雑誌

Cytometry research : 日本サイトメトリー学会機関誌 (ISSN:09166920)

巻号頁・発行日

vol.20, no.2, pp.27-33, 2010-09-25

参考文献数

6

被引用文献数

1

---

<p>Aim. We are in charge of the central diagnosis and cell preservation as a part of childhood acute lymphoblastic leukemia treatment study in Tokyo children's cancer study group. It is necessary to diagnose with a minimal quantity of specimen, to preserve leukemic cells effectively as possible. On the other hand, recent progress in multi-color flow cytometry enable to analyze cell marker of leukemia in more detail. We therefore we intended to perform a immuno-phenotypic diagnosis of childhood acute lymphoblastic leukemia by nine-color analysis with digital flow cytometer.</p><p>Methods. We examined cell markers of childhood acute lymphoblastic leukemia cells by nine-color analysis using digital flow cytometers. We decided the combination of the monoclonal antibodies for nine color analysis based on the recommendation of Japan Pediatric Leukemia/Lymphoma Study Group using commercially available fluorescence-laveled antibodies.</p><p>Results. Nine colors that we used in this study were fluorescein isothyocyanate, phycoerythrin (PE), phycoerythrin-Texas Red, Peridinin Chlorophyll Protein - cyanin (Cy) 5.5, PE-Cy7, allophycocyanin (APC), APC-Cy7, Pacific Blue, and Cascade Yellow. Using list mode compensation, nine color marker analysis of childhood leukemia was easy to perform.</p><p>Discussion. Although several problems need to be solved are present, nine-color analysis using digital flow cytometer is useful to obtain precise information of antigen expression as well as save precious specimen of childhood acute lymphoblastic leukemia.</p>

著者

永井 健治 宮脇 敦史

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.13, no.1, pp.1-10, 2003-06-20 (Released:2017-07-11)

参考文献数

30

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The discovery and development of green fluorescent protein (GFP) from the jellyfish Aequorea victoria and,more recently red fluorescent protein (dsRed) from the sea anemone Discosoma striata, have revolutionized our ability to study biological function such as protein localization, dynamics and interactions in living cells. The usefulness of GFPs derives from the finding that the fluorescent property of GFPs requires no other cofactor: The fluorophore forms spontaneously from the cyclization of the peptide backbone. Although GFPs promise high sensitivity and great versatility for biological applications, they sometimes send irresponsible signals because of the properties such as their sensitivity to pH and chloride fluctuations and photobleaching. Therefore, GFP users should be careful in interpreting the fluorescent signals from GFP variants to obtain reliable biological data. On the other hand, by using the characteristics, a wide range of application could be done. In this review, we will present 1) an overview of the physicochemical properties of GFPs, and 2) the valuable applications of GFP for biological research, including circularly permuted GFP technology and GFP-based fluorescence resonance energy transfer technique as well as their potential in a variety of biological sensors.

著者

宮本 京子 清島 久美 亀井 美沙 小方 由美子 前田 裕亮 前田 高宏

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.29, no.2, pp.21-27, 2019-11-25 (Released:2019-11-25)

参考文献数

14

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Progress has been made in the treatment of multiple myeloma (MM), and a series of novel therapeutic agents, including antibody-based drugs such as elotuzumab and daratumumab, are available in the clinic. While fl ow cytometry (FCM) is a major method for MM diagnosis and evaluation of therapeutic effects, detecting MM cells after antibodybased therapies is challenging, as antibodies used for FCM sometimes recognize the same epitopes that are targeted by the therapeutic ones. As a result, FCM could fail to detect true MM clones. In this study, we examined the effi cacy and accuracy of the FCM-based diagnostic methods using an antibody targeting multiple epitopes of CD38 (CD38ME) and intracellular p63 as well as those targeting CD138 and CD38high. When we defi ned MM cells using antibodies against CD38ME and intracellular p63, proportions of MM cells were highly correlated with those defined by the conventional FCM methods using anti-CD38high and / or CD138 antibodies (r2 = 0.9967-0.9991). Interestingly, expression levels of CD38high and CD138 were signifi cantly low in MM cells obtained from antibody-treated individuals. In contrast, MM clones were accurately detected using antibodies against CD38ME and intracellular p63. Our data suggest that extra caution should be taken when MM cells obtained from patients treated with antibody-based therapies were evaluated by FCM. We propose that antibodies targeting CD38ME and/or intracellular p63 should be included in the antibody mixture for FCM-based detection of MM cells.

著者

板野 直樹 木全 弘治

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.12, no.2, pp.1-6, 2002-12-25 (Released:2017-07-11)

参考文献数

19

---

Rapid advances in diagnostic and therapeutic techniques have greatly improved cancer prognosis. How ever, even at present, it is difficult to predict cancer metastasis, and the elucidation of the mechanism of can cer metastasis and its therapeutic application have just begun. Cancer progresses as a complex phenome non involving various life phenomena such as the abnormal proliferation of cells, their acquisition of motility,degradation of extracellular matrices, and changes in intercellular adhesiveness. The interaction of a very wide diversity of molecules acting inside and outside the cells controls the complex behaviors of cancer cells. In particular, molecules acting outside the cells are thought to directly specify the malignant characteristics of cancer cells; therefore, investigators are attempting to prevent cancer cell invasion and metastasis by inhibit ing their functions. The extracellularly acting molecules include a group of molecules, such as fibronectin and laminin, which interact with each other or with other molecules to form matrices and regulate tissue formation and cell functions. In general, extracellular matrix molecules such as collagen are thought to block the inva sion and metastasis of cancer cells. Conversely, hyaluronan, which is another matrix component and large sugar chain molecule, is synthesized, formed into matrices in many cancer tissues such as breast and colon cancers and gliomas, and is closely related to cancer progression. This paper focuses on hyaluronan and its matrix formation, gives an overview of studies aimed at elucidating tumor progression, and refers to recent trends towards blocking cancer invasion and metastasis by targeting the biosynthesis of hyaluronan.

著者

片岡 圭亮 小川 誠司

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.26, no.2, pp.15-20, 2016

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<p>PD-1/PD-L1 immune checkpoint pathway, which maintains self-tolerance and limits collateral tissue damage in a physiological state, can be co-opted by cancer cells to evade immune surveillance. Immune checkpoint inhibitors, such as anti-PD-1 and anti-PD-L1 antibodies, can unleash anti-tumor immunity and mediate durable cancer regression. However, the complex biology of this pathway has not been fully elucidated, including the genetic basis of PD-L1 upregulation in cancer. Recently, we have identified a novel genetic mechanism of immune evasion associated with structural variations (SVs) disrupting the 3'-untranlated region (UTR) of the <i>PD-L1</i> gene in a variety of cancers. Despite a large diversity in SV type, these SVs are invariably associated with a marked increase of PD-L1 expression, which promotes tumor progression and immune escape. Here we present an overview of PD-1/PD-L1 immune checkpoint blockade therapy, and highlight the genetic mechanisms of PD-L1 activation, including copy number amplification, promoter replacement, as well as 3'-UTR disruption, with a special focus on their relevance as a biomarker.</p>

著者

上田 舞 高島 康弘

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.27, no.1, pp.19-24, 2017-05-25 (Released:2017-05-31)

参考文献数

20

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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.

著者

林 真人 金 賢徹 寺薗 英之 服部 明弘 安田 賢二 大津 敬 宮城 洋平 荒尾 徳三 西尾 和人

出版者

日本サイトメトリー学会

雑誌

サイトメトリーリサーチ (ISSN:09166920)

巻号頁・発行日

vol.21, no.2, pp.1-6, 2011-10-25 (Released:2017-06-12)

参考文献数

13

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転移性がんの治療方針の策定や転移メカニズムの解明の為には,CTC を採血サンプル中から出来るだけ損傷を与えずに取りこぼし無く回収し,回収したCTC の生理学的性質や遺伝子変異を迅速に計測する技術が必要となる。 本稿では非浸襲的で高感度なCTC 分離精製法開発の取り組みについて概観した後,我々が開発を進めている画像認識型セルソーターシステムと超高速PCR のCTC 分離精製,遺伝子解析への応用可能性について論じた。

著者

塩沢 裕介 北村 紀子 竹野内 寿美 田口 智子 大喜多 肇 林 泰秀 小原 明 花田 良二 土田 昌宏 藤本 純一郎 清河 信敬

雑誌

Cytometry research = 日本サイトメトリー学会機関誌 (ISSN:09166920)

巻号頁・発行日

vol.16, no.2, pp.11-17, 2006-09-25

被引用文献数

2

著者

中村 有里

雑誌

Cytometry research : 日本サイトメトリー学会機関誌 (ISSN:09166920)

巻号頁・発行日

vol.9, no.2, pp.49-58, 1999-12-25

被引用文献数

1