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    T-Cell Xtend Technical Information

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    danang setiawan

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    © All Rights Reserved
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    T-Cell Xtend Technical Information

    Uploaded by

    danang setiawan
    23 views4 pages

    Original Title

    T-cell Xtend Technical Information

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    23 views4 pages

    T-Cell Xtend Technical Information

    Uploaded by

    danang setiawan

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 4

    T-Cell Xtend ™

    Technical information

    References:
    1. Oxford Immunotec. Data on File, 2008.
     chmielau J, Finn OJ. Activated granulocytes and granulocyte-derived H202 are the underlying mechanism of
    2. S
    suppression of T-cell function in advanced cancer patients. Cancer Research 2001; 61 4756-4760.
    3. M
     almberg KJ, Arulampalam V, Ishihara F, et al. Inhibition of activated/memory (CD45RO+) T cells by oxidative
    stress associated with block of NF-KB activation. J Immunol. 2001.167; 2595-2601.

    Contact Oxford Immunotec for further information.


    T-SPOT, T-Cell Xtend and the Oxford Immunotec logo are trademarks of Oxford Immunotec Ltd.
    © Oxford Immunotec Ltd. 2009. “All rights reserved”.
    Oxford Immunotec Limited, 94C Milton Park, Abingdon, Oxon OX14 4RY, UK
    Tel: +44 (0)1235 442780 Fax: +44 (0)1235 442781 Email: [email protected]
    www.oxfordimmunotec.com
    *FICOLL and FICOLL-PAQUE are trademarks of GE Healthcare companies.

    TI-TTK.610-UK-VI 030609
    The T-SPOT®.TB assay functions optimally with whole
    blood stored for up to 8 hours, but what happens if
    blood is stored for longer?
    result in a decrease in spot counts and an increase in non-specific background in
    – Performing the T-SPOT.TB assay on blood stored for more than 8 hours may

    assay wells

    FICOLL-PAQUE*, the majority of the red blood cells and granulocytes are pelleted
    – When peripheral blood mononuclear cells (PBMCs) are separated using

    on the basis of density

    – When blood is stored for longer than 8 hours, granulocytes can become activated
    causing a decrease in their density1. As a result, the granulocytes can contaminate
    the enriched PBMC layer

    Effect of blood storage on T-SPOT.TB performance


    55 12
    50 11
    45 10
    40 9
    35 8

    Spot Counts

    Spot Counts
    7
    30
    6
    25
    5
    20
    4
    15 3
    10 2
    5 1
    0 0
    <8 HOURS 16-18 HOURS <8 HOURS 24 HOURS

    
    – Granulocytes are known to become activated in some disease states (e.g. cancer)2,3

    – Activated granulocytes can release PBMCs


    pre-stored granules2, which may cause
    oxidative stress to lymphocytes in the
    PBMC layer. This may reduce cell
    Granulocytes
    viability and, more specifically, the
    ability of cells to release interferon-
    gamma FICOLL

    – The presence of granulocytes may


    also result in non-specific background,
    affecting results with stored blood Erythrocytes

    With stored blood granulocytes can appear


    in the PBMC layer
    The T-Cell Xtend reagent allows the T-SPOT.TB assay The T-Cell Xtend reagent removes unwanted
    to be performed on blood samples stored for up to granulocytes from the sample
    32 hours without compromising accuracy
    Use of the T-Cell Xtend reagent has been demonstrated to reduce the presence of
    granulocytes in the PBMC layer, thus allowing prolonged storage of blood samples for
    up to 32 hours before performing the T-SPOT.TB assay

    60

    50

    40

    30

    %CD66b
    20

    The T-Cell Xtend reagent is an antibody complex which recognises CD66b, a specific 10
    cell surface marker of granulocytes, and cross-links the granulocytes to red blood cells

    PBMCs 0-8 24 32 48 72 0-8 24 32 48 72


    -10
    Time (Hrs)

    FICOLL
    Granulocyte contamination of Granulocyte contamination
    PBMC layer increases with of PBMC layer is minimal
    blood storage time with increased blood storage
    time and addition of the
    T-Cell Xtend reagent
    Granulocytes

    Erythrocytes

    This cross-linking increases the density of granulocytes so they pellet when applied to
    a density gradient
    The T-Cell Xtend reagent is added to blood samples Benefits of using the T-Cell Xtend reagent
    in the laboratory, immediately before commencing the
    T-SPOT.TB assay
    Equivalent accuracy to standard T-SPOT.TB results but with added
    T-Cell Xtend may also be used with flexibility
    Leucosep tubes to simplify the FICOLL
    separation procedure – Samples can be collected at a distance from the testing laboratory

    – Samples can be collected throughout the day or stored overnight

    – Laboratory can perform T-SPOT.TB on blood samples received on


    2 consecutive days at the same time

    – Enables the use of cost-effective, overnight delivery service

    – Removal of activated granulocytes may improve performance of


    the T-SPOT.TB assay in certain disease states (e.g. cancer)

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