Fluorospot assay: methodological analysis.

The conventional enzyme-linked immunospot (ELISPOT) technique detects only one secreted cytokine, which constitutes a major pitfall for the accurate characterization of the various T-cell subpopulations. We have therefore developed a fluorospot assay, which is a modification of the ELISPOT and is based on the use of multiple fluorescent-labeled anticytokines detection antibodies. A special automated ELISPOT reader consisting of a light microscope with incident fluorescence illumination and an integrating digital color camera has been adapted for this technique.

Optimization of an elispot assay to detect cytomegalovirus-specific CD8+ T lymphocytes.

Various arguments suggest that CD8+ T lymphocytes play a major role in the control of cytomegalovirus (CMV) infection. The detection of CMV-specific CD8+ T cells may therefore provide additional information about CMV virus detection to predict the risk of development of CMV disease, especially in immunodepressed transplant recipients. We compared and tested various experimental conditions to optimize an enzyme-linked immunospot assay (Elispot) assay for the detection of CMV-specific CD8+ T lymphocytes.

A Fluorospot assay to detect single T lymphocytes simultaneously producing multiple cytokines.

Various subpopulations of T lymphocytes-i.e. Type 1, Type 2, Tr1 T cells-play a major role in the homeostasis of the immune system and in the pathogenesis of many inflammatory and auto-immune diseases.

[Cytotoxic T lymphocytes: role in immunosurveillance and in immunotherapy].

Experiments in mice and recent human clinical studies have clearly shown the contribution of CD8+ T lymphocyte in the control of tumor development. CD8+ T lymphocytes are a constitutive component of the immune response during the development of cancer. In murine models, the efficiency of various cancer vaccines mainly depends on their ability to induce CD8+ T lymphocytes.