Journal
BIOCHEMICAL PHARMACOLOGY
Volume 80, Issue 12, Pages 2087-2091Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.bcp.2010.09.004
Keywords
Cytotoxic T lymphocytes; Cell migration; Real-time imaging; Immunotherapy; Cytotoxicity assay
Categories
Funding
- DFG [FR1155/6-3]
- EU [ENCITE HEALTH TH-15-2008-208142]
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Cytotoxic T lymphocytes (CTL) mediate antigen- and cell-cell contact dependent killing of target cells, such as cancer cells and virus-infected cells. In vivo, this process requires the active migration of OM towards and away from target cells. We here describe an organotypic 3D collagen matrix assay to monitor CTL migration together with CTL-mediated killing of target cells. The assay supports both, time-lapse microscopy of killing dynamics as well as population analysis of killing after matrix digestion and flow cytometry. The assay was used to assess the detrimental effect of cyclosporine A (CsA) present during CTL activation, which caused an inhibition of CTL-target cell conjugation and strongly impaired CTL-mediated killing, particularly at low effector-target ratios. Thus, the organotypic assay is useful to monitor spatiotemporal control mechanisms of cytotoxic immune effector functions. (C) 2010 Elsevier Inc. All rights reserved.
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