Serum-free culture medium and IL-7 costimulation increase the sensitivity of ELISpot detection. - Archive ouverte HAL Access content directly
Journal Articles Journal of Immunological Methods Year : 2008

Serum-free culture medium and IL-7 costimulation increase the sensitivity of ELISpot detection.

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Abstract

The identification of parameters maximizing detection sensitivity in ELISpot assays is important to transfer this technology into the clinical setting for identifying rare Ag-specific CD8(+) T cells. We have therefore considered human IFN-gamma CD8(+) T cell responses against viral epitopes to analyze different variables which could be critical during the epitope-specific stimulation period. Two parameters were found to greatly enhance detection sensitivity (i.e., to specifically increase epitope-driven signal while keeping background noise to a minimum): use of human serum-free vs. serum-supplemented culture medium (2.4-fold median increase) and addition of low dose IL-7 (1.5-fold increase). Incorporating both of these parameters into the ELISpot procedure proved capable of greatly amplifying (35.1-fold increase) the low grade CD8(+) T cell responses directed against beta-cell epitopes of type 1 diabetes patients, as compared to a previously optimized procedure using human serum-supplemented medium and low dose IL-2. Implementation of this ELISpot procedure should expedite development of "immune staging" protocols for autoimmune as well as tumor and infectious diseases.
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Dates and versions

inserm-00266553 , version 1 (26-03-2008)

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Emanuela Martinuzzi, Matthieu Scotto, Emmanuelle Enée, Vedran Brezar, Jean-Antoine Ribeil, et al.. Serum-free culture medium and IL-7 costimulation increase the sensitivity of ELISpot detection.. Journal of Immunological Methods, 2008, 333 (1-2), pp.61-70. ⟨10.1016/j.jim.2008.01.003⟩. ⟨inserm-00266553⟩
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