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  1. AU="Ruliang Li"
  2. AU="Gilchriese, M G D"
  3. AU="Rist, Andreas"
  4. AU="Katznelson, Andrew" AU="Katznelson, Andrew"
  5. AU="Solís-Martínez, Obed"
  6. AU="Dumitrescu, Florentina"
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  1. Article ; Online: A Positive Control for Detection of Functional CD4 T Cells in PBMC

    Annemarie Schiller / Ting Zhang / Ruliang Li / Andrea Duechting / Srividya Sundararaman / Anna Przybyla / Stefanie Kuerten / Paul V. Lehmann

    Cells, Vol 6, Iss 4, p

    The CPI Pool

    2017  Volume 47

    Abstract: Testing of peripheral blood mononuclear cells (PBMC) for immune monitoring purposes requires verification of their functionality. This is of particular concern when the PBMC have been shipped or stored for prolonged periods of time. While the CEF ( ... ...

    Abstract Testing of peripheral blood mononuclear cells (PBMC) for immune monitoring purposes requires verification of their functionality. This is of particular concern when the PBMC have been shipped or stored for prolonged periods of time. While the CEF (Cytomegalo-, Epstein-Barr and Flu-virus) peptide pool has become the gold standard for testing CD8 cell functionality, a positive control for CD4 cells is so far lacking. The latter ideally consists of proteins so as to control for the functionality of the antigen processing and presentation compartments, as well. Aiming to generate a positive control for CD4 cells, we first selected 12 protein antigens from infectious/environmental organisms that are ubiquitous: Varicella, Influenza, Parainfluenza, Mumps, Cytomegalovirus, Streptococcus, Mycoplasma, Lactobacillus, Neisseria, Candida, Rubella, and Measles. Of these antigens, three were found to elicited interferon (IFN)-γ-producing CD4 cells in the majority of human test subjects: inactivated cytomegalo-, parainfluenza-, and influenza virions (CPI). While individually none of these three antigens triggered a recall response in all donors, the pool of the three (the ‘CPI pool’), did. One hundred percent of 245 human donors tested were found to be CPI positive, including Caucasians, Asians, and African-Americans. Therefore, the CPI pool appears to be suitable to serve as universal positive control for verifying the functionality of CD4 and of antigen presenting cells.
    Keywords PBMC quality assessment ; PBMC cryopreservation ; CD4 cell function ; ELISPOT ; ImmunoSpot ; immune monitoring ; Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2017-12-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring Cell-Mediated Cytotoxicity

    Anna Welter / Srividya Sundararaman / Ruliang Li / Ting Zhang / Alexey Y. Karulin / Alexander Lehmann / Villian Naeem / Diana R. Roen / Stefanie Kuerten / Paul V. Lehmann

    Cells, Vol 7, Iss 5, p

    2018  Volume 35

    Abstract: One of the primary effector functions of immune cells is the killing of virus-infected or malignant cells in the body. Natural killer (NK) and CD8 effector T cells are specialized for this function. The gold standard for measuring such cell-mediated ... ...

    Abstract One of the primary effector functions of immune cells is the killing of virus-infected or malignant cells in the body. Natural killer (NK) and CD8 effector T cells are specialized for this function. The gold standard for measuring such cell-mediated cytolysis has been the chromium release assay, in which the leakage of the radioactive isotope from damaged target cells is being detected. Flow cytometry-based single cell analysis of target cells has recently been established as a non-radioactive alternative. Here we introduce a target cell visualization assay (TVA) that applies similar target cell staining approaches as used in flow cytometry but based on single cell computer image analysis. Two versions of TVA are described here. In one, the decrease in numbers of calcein-stained, i.e., viable, target cells is assessed. In the other, the CFSE/PI TVA, the increase in numbers of dead target cells is established in addition. TVA assays are shown to operate with the same sensitivity as standard chromium release assays, and, leaving data audit trails in form of scanned (raw), analyzed, and quality-controlled images, thus meeting requirements for measuring cell-mediated cytolysis in a regulated environment.
    Keywords TVA ; NK measurement ; ADCC ; CD8 cells ; multi-color single cell imaging ; CFR-Part 11 compliance ; audit trails ; cellular assay validation ; immune monitoring ; Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2018-04-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

    Full text online

    More links

    Kategorien

    Inter-library loan at ZB MED

    Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

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