Giuseppe
Emeritus
Standardization and validation of assays determining cellular immune responses against influenza. (Vaccine, abstract, edited)
[Source: US National Library of Medicine, LINK. EDITED.]
Vaccine. 2010 Mar 2. [Epub ahead of print]
Standardization and validation of assays determining cellular immune responses against influenza.
Gijzen K, Liu WM, Visontai I, Oftung F, van der Werf S, Korsvold GE, Pronk I,Aaberge IS, T?ttő A, Jankovics I, Jankovics M, Gentleman B, McElhaney JE,Soethout EC. - Vaccine Research, Netherlands Vaccine Institute (NVI), Bilthoven, TheNetherlands.
Influenza vaccine efficacy does not always correlate with humoral immune responses. Recent reports indicate that the cellular immune response also contributes to protection, however robust assays are lacking. We standardized and validated assays for detection of human influenza-specific cellular responses in four international laboratories. The production of granzyme B as marker of T cell-mediated cytotoxicity and release of Th1 and Th2 cytokines were evaluated. The granzyme B and cytokine assays were specific, accurate, precise, and robust. Replicate stimulations with PBMC from the same donors showed an intra-laboratory robustness (coefficient of variation) for quantitation of granzyme B of 33% and for cytokines - including IFN-gamma, TNF-alpha, IL-2, IL-10, IL-4, IL-13, GM-CSF and including the log IFN-gamma/IL-10 ratio - of 52%. The inter-laboratory robustness for detection of granzyme B was 29% and for detection of all cytokines was 49%. The assays can now be used for determining cell-mediated immunity and explored as correlates of protection. Moreover, the precision and robustness of these cellular assays allow the reliable detection of cellular responses even in small study populations.
Copyright ? 2010 Elsevier Ltd. All rights reserved.
PMID: 20206285 [PubMed - as supplied by publisher]
Vaccine. 2010 Mar 2. [Epub ahead of print]
Standardization and validation of assays determining cellular immune responses against influenza.
Gijzen K, Liu WM, Visontai I, Oftung F, van der Werf S, Korsvold GE, Pronk I,Aaberge IS, T?ttő A, Jankovics I, Jankovics M, Gentleman B, McElhaney JE,Soethout EC. - Vaccine Research, Netherlands Vaccine Institute (NVI), Bilthoven, TheNetherlands.
Influenza vaccine efficacy does not always correlate with humoral immune responses. Recent reports indicate that the cellular immune response also contributes to protection, however robust assays are lacking. We standardized and validated assays for detection of human influenza-specific cellular responses in four international laboratories. The production of granzyme B as marker of T cell-mediated cytotoxicity and release of Th1 and Th2 cytokines were evaluated. The granzyme B and cytokine assays were specific, accurate, precise, and robust. Replicate stimulations with PBMC from the same donors showed an intra-laboratory robustness (coefficient of variation) for quantitation of granzyme B of 33% and for cytokines - including IFN-gamma, TNF-alpha, IL-2, IL-10, IL-4, IL-13, GM-CSF and including the log IFN-gamma/IL-10 ratio - of 52%. The inter-laboratory robustness for detection of granzyme B was 29% and for detection of all cytokines was 49%. The assays can now be used for determining cell-mediated immunity and explored as correlates of protection. Moreover, the precision and robustness of these cellular assays allow the reliable detection of cellular responses even in small study populations.
Copyright ? 2010 Elsevier Ltd. All rights reserved.
PMID: 20206285 [PubMed - as supplied by publisher]