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Published on: September 9, 2011
Correlation of human CD56+ cell cytotoxicity and IFN-gamma production
E G Derby1, V Reddy, E L Nelson
1NCI-Frederick Cancer Research and Development Center, SAIC-Frederick, Frederick, MD 21702-1201, USA.
Insights
The Interferon-gamma ELISPOT assay reliably detects natural killer (NK) cell activity, correlating well with traditional chromium-51 release assays. This offers a valuable alternative for assessing cellular immune responses.
Area of Science:
- Immunology
- Cellular immunology
- Immunoassays
Background:
- Interferon-gamma (IFN-gamma) ELISPOT assays are increasingly used to assess cellular immune responses, particularly cytotoxic T lymphocyte (CTL) activity.
- Traditional chromium-51 (51Cr) release assays are a standard method for measuring natural killer (NK) cell activity.
Purpose of the Study:
- To compare the efficacy of the IFN-gamma ELISPOT assay with the traditional 51Cr release assay for detecting human NK cell activity.
- To identify the primary source of IFN-gamma production in peripheral blood mononuclear cells (PBMC) during NK cell assays.
Main Methods:
- Evaluation of freshly isolated and interleukin-2 (IL-2) activated peripheral blood mononuclear cells (PBMC).
- Comparison of IFN-gamma ELISPOT assay results with 51Cr release assay data.
- Identification of CD56-positive cells as the source of IFN-gamma signal.
Main Results:
- The IFN-gamma ELISPOT assay demonstrated a strong correlation with the 51Cr release assay in evaluating NK cell activity.
- CD56-positive cells were identified as the main producers of IFN-gamma when PBMC were tested against NK-sensitive targets using the ELISPOT assay.
Conclusions:
- The IFN-gamma ELISPOT assay is a reliable method for assessing human NK cell activity.
- ELISPOT assays provide a viable alternative to 51Cr release assays for evaluating NK cell-mediated immune responses.
Abstract:
The use of an IFN-gamma ELISPOT assay to evaluate cellular immune responses has gained increasing popularity, especially as a surrogate measure for cytotoxic T lymphocyte (CTL) responses. We have compared the IFN-gamma ELISPOT assay and the traditional(51)Cr release assay for detection of human natural killer (NK) cell activity. The cell populations used for evaluation of these assays included freshly isolated and IL-2-activated peripheral blood mononuclear cells (PBMC). CD56-positive cells were demonstrated to be the primary source of the IFN-gamma signal when PBMC were evaluated with NK-sensitive targets in the IFN-gamma ELISPOT assay. IFN-gamma ELISPOT and(51)Cr release assays showed excellent correlation suggesting that NK activity can be reliably evaluated with methods other than the traditional(51)Cr release assays.
