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11-color, 13-parameter flow cytometry: Identification of human naive T cells by phenotype, function, and T-cell receptor diversity

Nature Medicine volume 7, pages 245–248 (2001)Cite this article

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Recent findings illustrate that diseases are often accompanied by changes in the numbers or function of `fine' lymphocyte subsets, even if changes in the bulk lymphocyte populations are not evident. In some cases, such changes may provide powerful prognostic and diagnostic information. For example, Giorgi et al.1 showed that the number of activated CD8 T cells (CD38+/HLA-DR+), but not the number of total CD8 T cells, was a better predictor of progression to AIDS than the commonly-used CD4 T-cell count. And, by accurately dividing T cells into naive and memory, we demonstrated that progression of HIV disease was accompanied by the loss of both CD4 and CD8 naive T cells2,3—a finding that could not be explained by then-popular models of cell loss by viral cytolysis.

These examples illustrate our increasing awareness of the complexity of the immune system. As many diseases are associated with expansions or reductions in major leukocyte subsets (for example, anemias, neutropenias, cancers and AIDS), it should come as no surprise that changes in representation (absolute number or percentage) of fine subsets of lymphocytes are also correlated with disease. Identifying such changes has been difficult because traditional three-color flow cytometric analyses can only resolve major lineages (B, T, natural killer, monocyte or neutrophil), or, at best, resolve a few subsets of any single lineage.

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Figure 1: 13-parameter, 11-color PFC accurately defines naive T cells. Peripheral blood mononuclear cells from one donor were stained with one 11-color staining combination (Table 1) and analyzed by PFC.
Figure 2: The use of multiple markers increases the accuracy of naive cell determinations.
Figure 3: Cells discordant for naive markers express IFN-γ upon stimulation.
Figure 4: CD8 T cells discordant for naive markers display a restricted Vβ repertoire and are therefore memory cells.

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Acknowledgements

This work was supported by grants from the NIH (CA-42509-14, CA-81543-02), and from the University of California University-wide AIDS Research Program (F97-ST-044).

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Authors and Affiliations

  1. Vaccine Research Center, National Institutes of Health, 40 Convent Drive, Bethesda, Maryland, USA

    Stephen C. De Rosa & Mario Roederer

  2. Department of Genetics, Stanford University School of Medicine, Stanford, California, USA

    Leonard A. Herzenberg & Leonore A. Herzenberg

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  1. Stephen C. De Rosa
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Correspondence to Mario Roederer.

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De Rosa, S., Herzenberg, L., Herzenberg, L. et al. 11-color, 13-parameter flow cytometry: Identification of human naive T cells by phenotype, function, and T-cell receptor diversity. Nat Med 7, 245–248 (2001). https://doi.org/10.1038/84701

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