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The CD107 mobilization assay: viable isolation and immunotherapeutic potential of tumor-cytolytic NK cells

Leukemia volume 19, pages 707–709 (2005)Cite this article

In this issue of Leukemia, Penack et al1 report on the identification and isolation of tumor-cytolytic natural killer (NK) cells on the basis of their CD107 surface expression. The CD107 assay constitutes a promising new tool for characterization and possibly immunotherapeutic use of tumor-specific NK cells. Several techniques are already available to measure cytotoxicity of T and NK cells. The classical cytotoxicity assay, measuring release of chromium 51Cr (or more recently of nonradioactive fluorescent dyes)2 from lysed target cells, is a sensitive and reliable method for assessing cytotoxicity of bulk NK cells, but gives no quantitative information on the frequency of cytotoxic effector cells.3 A more direct and quantitative assessment of cytolytic cells became possible via ELISPOT analysis of granzyme B release.4 Recently, Betts and colleagues developed a simple flow cytometric assay, which enables the identification and isolation of viable cytolytic T cells, based on surface mobilization of CD107.5 Peter Lee's laboratory applied the CD107 mobilization assay for the specific isolation and subsequent ex vivo expansion of tumor-specific cytolytic T cells from peptide-vaccinated melanoma patients.6 Now, Penack et al1 demonstrate that the CD107 mobilization assay can equally well be utilized for isolation of tumor-cytolytic NK cells.

The major weapon of NK cells as well as cytotoxic T-lymphocytes (CTL) are specialized secretory lysosomes, the lytic granules.7 When a target cell is identified, the content of these granules is released by exocytosis into the immunological synapse formed between killer and target cell, eventually leading to the induction of apoptosis.8, 9 The ammunition stored in these granules includes the serine proteases granzyme A and B, the membrane-disrupting protein perforin, and the antimicrobial lytic molecule granulysin.10 The molecular events leading to delivery of the lethal hit are only starting to get unraveled. Inside the granules, perforin and granzymes are complexed with the proteoglycan serglycin.11 Upon activation, the multimeric complexes are exocytosed and subsequently enter the target cell by yet unknown mechanisms. Serglycin acts as a carrier molecule for granzyme and might also facilitate the uptake of granzyme into the target cell.11 Earlier studies had suggested that internalization of granzyme is based on the pore-forming capacity of perforin, which would enable passive diffusion of granzymes into the target cell. However, recent results suggest that, rather than drilling holes into the target cell plasma membrane, perforin acts subsequent to entering the target cell by enabling release of lytic complexes from endocytic vesicles of the target cell, where the granzymes would otherwise get trapped.7, 11 Upon cytosolic delivery of granzymes, cell death is then induced through triggering of different, caspase-independent (granzyme A and B) as well as caspase-dependent (granzyme B) apoptotic pathways.10

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Acknowledgements

This work was supported by a grant from the Deutsche Krebshilfe.

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  1. Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University Düsseldorf, Düsseldorf, Germany

    M Uhrberg

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Uhrberg, M. The CD107 mobilization assay: viable isolation and immunotherapeutic potential of tumor-cytolytic NK cells. Leukemia 19, 707–709 (2005). https://doi.org/10.1038/sj.leu.2403705

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