A high-throughput single-cell analysis of human CD8(+) T cell functions reveals discordance for cytokine secretion and cytolysis

Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
The Journal of clinical investigation (Impact Factor: 13.22). 10/2011; 121(11):4322-31. DOI: 10.1172/JCI58653
Source: PubMed


CD8+ T cells are a key component of the adaptive immune response to viral infection. An inadequate CD8+ T cell response is thought to be partly responsible for the persistent chronic infection that arises following infection with HIV. It is therefore critical to identify ways to define what constitutes an adequate or inadequate response. IFN-γ production has been used as a measure of T cell function, but the relationship between cytokine production and the ability of a cell to lyse virus-infected cells is not clear. Moreover, the ability to assess multiple CD8+ T cell functions with single-cell resolution using freshly isolated blood samples, and subsequently to recover these cells for further functional analyses, has not been achieved. As described here, to address this need, we have developed a high-throughput, automated assay in 125-pl microwells to simultaneously evaluate the ability of thousands of individual CD8+ T cells from HIV-infected patients to mediate lysis and to produce cytokines. This concurrent, direct analysis enabled us to investigate the correlation between immediate cytotoxic activity and short-term cytokine secretion. The majority of in vivo primed, circulating HIV-specific CD8+ T cells were discordant for cytolysis and cytokine secretion, notably IFN-γ, when encountering cognate antigen presented on defined numbers of cells. Our approach should facilitate determination of signatures of functional variance among individual effector CD8+ T cells, including those from mucosal samples and those induced by vaccines.

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Available from: Navin Varadarajan, Oct 01, 2015
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    • "First of all, ICS does not retain cell viability, and thus, recovery of cells for clonal expansion is difficult. In addition, since a protein transport inhibitor is added to retain the cytokines within the cell, it is difficult to determine whether the detected cytokines are intracellular, secreted or membrane-bound [72]. Moreover, dynamic monitoring of an individual cell is difficult since the secretions of the cells upstream may affect others. "
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    PLoS ONE 01/2014; 9(1):e85675. DOI:10.1371/journal.pone.0085675 · 3.23 Impact Factor
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    • "While flow cytometry has traditionally been used to determine single cell phenotypes, it cannot provide continuous measurements of proteins in the same individual cells over time7. Recently, the development of microfluidic technology enabled analysis of cell phenomena on the single cell level8910. The reported platforms, mostly based on micro well technologies, do not provide a controllable microenvironment for cell-cell interaction, however, as no reagent mixing is provided. "
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