Abstract
Flow cytometric measurement of intracellular cytokines in T cells exposed to antigen is a widely used method for quantification of an antigen-specific T-cell response. As the frequency of antigen-specific T cells is often very low, any improvement in signal to noise ratio is of great importance. Thus, in this study, the ability of antigen-pulsed dendritic cells (DCs) to increase the number of antigen-specific, interferon-gamma (IFN-gamma)-producing CD4+ T cells measurable both in fresh peripheral blood and in reconstituted frozen blood mononuclear cell (MNC) samples was evaluated. Cytomegalovirus (CMV) was used as antigen in a 10 h assay, using cells from both CMV-seropositive and -seronegative donors. When reconstituted frozen samples were analysed, the general response towards CMV lysate in CMV-seropositive donors was 23-86% lower compared to the corresponding fresh blood samples. Antigen-pulsed DCs could not improve the sensitivity of the intracellular cytokine-detection assay when fresh peripheral blood samples were used. Interestingly, however, the addition of CMV lysate-pulsed DCs to cryopreserved MNC samples substantially increased the frequency of specifically induced IFN-gamma-producing cells to a level comparable to the frequency found in the corresponding fresh blood samples.
Original language | English |
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Journal | Scandinavian Journal of Immunology |
Volume | 59 |
Issue number | 1 |
Pages (from-to) | 88-96 |
Number of pages | 9 |
ISSN | 0300-9475 |
Publication status | Published - Jan 2004 |
Externally published | Yes |
Keywords
- Adult
- Animals
- Blood Chemical Analysis
- CD4-Positive T-Lymphocytes
- Cytomegalovirus
- Cytomegalovirus Infections
- Dendritic Cells
- Female
- Flow Cytometry
- Humans
- Immunoglobulin G
- Interferon-gamma
- Intracellular Fluid
- Lymphocyte Activation
- Male
- Middle Aged
- Specimen Handling
- Journal Article
- Research Support, Non-U.S. Gov't