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  1. Article: Highly multiplexed quantitation of gene expression on single cells

    Dominguez, Maria H / Chattopadhyay, Pratip K / Ma, Steven / Lamoreaux, Laurie / McDavid, Andrew / Finak, Greg / Gottardo, Raphael / Koup, Richard A / Roederer, Mario

    Journal of immunological methods. 2013 May 31, v. 391, no. 1-2

    2013  

    Abstract: Highly multiplexed, single-cell technologies reveal important heterogeneity within cell populations. Recently, technologies to simultaneously measure expression of 96 (or more) genes from a single cell have been developed for immunologic monitoring. Here, ...

    Abstract Highly multiplexed, single-cell technologies reveal important heterogeneity within cell populations. Recently, technologies to simultaneously measure expression of 96 (or more) genes from a single cell have been developed for immunologic monitoring. Here, we report a rigorous, optimized, quantitative methodology for using this technology. Specifically: we describe a unique primer/probe qualification method necessary for quantitative results; we show that primers do not compete in highly multiplexed amplifications; we define the limit of detection for this assay as a single mRNA transcript; and, we show that the technical reproducibility of the system is very high. We illustrate two disparate applications of the platform: a “bulk” approach that measures expression patterns from 100 cells at a time in high throughput to define gene signatures, and a single-cell approach to define the coordinate expression patterns of multiple genes and reveal unique subsets of cells.
    Keywords detection limit ; gene expression ; genes ; messenger RNA ; monitoring
    Language English
    Dates of publication 2013-0531
    Size p. 133-145.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2013.03.002
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 795: Show issues Location:
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  2. Article ; Online: Highly multiplexed quantitation of gene expression on single cells.

    Dominguez, Maria H / Chattopadhyay, Pratip K / Ma, Steven / Lamoreaux, Laurie / McDavid, Andrew / Finak, Greg / Gottardo, Raphael / Koup, Richard A / Roederer, Mario

    Journal of immunological methods

    2013  Volume 391, Issue 1-2, Page(s) 133–145

    Abstract: Highly multiplexed, single-cell technologies reveal important heterogeneity within cell populations. Recently, technologies to simultaneously measure expression of 96 (or more) genes from a single cell have been developed for immunologic monitoring. Here, ...

    Abstract Highly multiplexed, single-cell technologies reveal important heterogeneity within cell populations. Recently, technologies to simultaneously measure expression of 96 (or more) genes from a single cell have been developed for immunologic monitoring. Here, we report a rigorous, optimized, quantitative methodology for using this technology. Specifically: we describe a unique primer/probe qualification method necessary for quantitative results; we show that primers do not compete in highly multiplexed amplifications; we define the limit of detection for this assay as a single mRNA transcript; and, we show that the technical reproducibility of the system is very high. We illustrate two disparate applications of the platform: a "bulk" approach that measures expression patterns from 100 cells at a time in high throughput to define gene signatures, and a single-cell approach to define the coordinate expression patterns of multiple genes and reveal unique subsets of cells.
    MeSH term(s) Animals ; DNA Primers ; Gene Expression Profiling/methods ; Gene Expression Regulation ; Genetic Markers ; High-Throughput Screening Assays/methods ; Humans ; Limit of Detection ; Linear Models ; Lymphocyte Activation/genetics ; Lymphocyte Subsets/immunology ; Macaca mulatta ; Multiplex Polymerase Chain Reaction ; RNA, Messenger/metabolism ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes/immunology
    Chemical Substances DNA Primers ; Genetic Markers ; RNA, Messenger
    Language English
    Publishing date 2013-03-13
    Publishing country Netherlands
    Document type Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2013.03.002
    Database MEDical Literature Analysis and Retrieval System OnLINE

    Full text online

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 795: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  3. Article ; Online: Superior T memory stem cell persistence supports long-lived T cell memory.

    Lugli, Enrico / Dominguez, Maria H / Gattinoni, Luca / Chattopadhyay, Pratip K / Bolton, Diane L / Song, Kaimei / Klatt, Nichole R / Brenchley, Jason M / Vaccari, Monica / Gostick, Emma / Price, David A / Waldmann, Thomas A / Restifo, Nicholas P / Franchini, Genoveffa / Roederer, Mario

    The Journal of clinical investigation

    2013  Volume 123, Issue 2, Page(s) 594–599

    Abstract: Long-lived memory T cells are able to persist in the host in the absence of antigen; however, the mechanism by which they are maintained is not well understood. Recently, a subset of human T cells, stem cell memory T cells (TSCM cells), was shown to be ... ...

    Abstract Long-lived memory T cells are able to persist in the host in the absence of antigen; however, the mechanism by which they are maintained is not well understood. Recently, a subset of human T cells, stem cell memory T cells (TSCM cells), was shown to be self-renewing and multipotent, thereby providing a potential reservoir for T cell memory throughout life. However, their in vivo dynamics and homeostasis still remain to be defined due to the lack of suitable animal models. We identified T cells with a TSCM phenotype and stem cell-like properties in nonhuman primates. These cells were the least-differentiated memory subset, were functionally distinct from conventional memory cells, and served as precursors of central memory. Antigen-specific TSCM cells preferentially localized to LNs and were virtually absent from mucosal surfaces. They were generated in the acute phase of viral infection, preferentially survived in comparison with all other memory cells following elimination of antigen, and stably persisted for the long term. Thus, one mechanism for maintenance of long-term T cell memory derives from the unique homeostatic properties of TSCM cells. Vaccination strategies designed to elicit durable cellular immunity should target the generation of TSCM cells.
    MeSH term(s) Animals ; Antigens, Viral ; Cytokines/biosynthesis ; Humans ; Immunologic Memory ; Immunophenotyping ; Lymphocyte Activation ; Macaca mulatta/immunology ; Macaca nemestrina/immunology ; Multipotent Stem Cells/immunology ; Simian Immunodeficiency Virus/immunology ; Species Specificity ; T-Lymphocyte Subsets/immunology
    Chemical Substances Antigens, Viral ; Cytokines
    Language English
    Publishing date 2013-01-02
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural
    ZDB-ID 3067-3
    ISSN 1558-8238 ; 0021-9738
    ISSN (online) 1558-8238
    ISSN 0021-9738
    DOI 10.1172/JCI66327
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Highly multiplexed quantitation of gene expression on single cells

    Dominguez, Maria H. / Chattopadhyay, Pratip K. / Ma, Steven / Lamoreaux, Laurie / McDavid, Andrew / Finak, Greg / Gottardo, Raphael / Koup, Richard A. / Roederer, Mario

    Journal of immunological methods

    Volume v. 391,, Issue no. 1

    Abstract: Highly multiplexed, single-cell technologies reveal important heterogeneity within cell populations. Recently, technologies to simultaneously measure expression of 96 (or more) genes from a single cell have been developed for immunologic monitoring. Here, ...

    Abstract Highly multiplexed, single-cell technologies reveal important heterogeneity within cell populations. Recently, technologies to simultaneously measure expression of 96 (or more) genes from a single cell have been developed for immunologic monitoring. Here, we report a rigorous, optimized, quantitative methodology for using this technology. Specifically: we describe a unique primer/probe qualification method necessary for quantitative results; we show that primers do not compete in highly multiplexed amplifications; we define the limit of detection for this assay as a single mRNA transcript; and, we show that the technical reproducibility of the system is very high. We illustrate two disparate applications of the platform: a “bulk” approach that measures expression patterns from 100 cells at a time in high throughput to define gene signatures, and a single-cell approach to define the coordinate expression patterns of multiple genes and reveal unique subsets of cells.
    Keywords monitoring ; messenger RNA ; detection limit ; genes ; gene expression
    Language English
    Document type Article
    ISSN 0022-1759
    Database AGRIS - International Information System for the Agricultural Sciences and Technology

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