Unit

UNIT 3.37 Isolation of Cytotoxic T Cell and NK Granules and Purification of Their Effector Proteins

  1. Jerome Thiery1,2,
  2. Michael Walch1,2,
  3. Danielle K. Jensen1,2,
  4. Denis Martinvalet1,2,
  5. Judy Lieberman1,2

Published Online: 1 JUN 2010

DOI: 10.1002/0471143030.cb0337s47

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Thiery, J., Walch, M., Jensen, D. K., Martinvalet, D. and Lieberman, J. 2010. Isolation of Cytotoxic T Cell and NK Granules and Purification of Their Effector Proteins. Current Protocols in Cell Biology. 47:3.37.1–3.37.29.

Author Information

  1. 1

    Program in Cellular and Molecular Medicine, Children's Hospital and Immune Disease Institute, Boston, Massachusetts

  2. 2

    Department of Pediatrics, Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 JUN 2010
  2. Published Print: JUN 2010

Abstract

Killer lymphocytes induce apoptosis by the release of cytotoxic mediators from specialized secretory lysosomes, called cytotoxic granules, into the immunological synapse formed with a cell targeted for elimination. Methods are presented here for isolating CTL and NK cell cytotoxic granules using cell disruption by nitrogen cavitation followed by continuous Percoll density gradient fractionation. Protocols are also given for purifying the key cytolytic molecules (perforin, granzyme A, granzyme B, and granulysin) from isolated cytotoxic granules by fast protein liquid chromatography. Curr. Protoc. Cell Biol. 47:3.37.1-3.37.29. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • cytotoxic granules;
  • CTL;
  • NK cells;
  • perforin;
  • granzyme;
  • granulysin