UNIT 3.15 Quantitation of Functional T Cells by Limiting Dilution
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Miller, R. A. 2001. Quantitation of Functional T Cells by Limiting Dilution. Current Protocols in Immunology. 35:III:3.15:3.15.1–3.15.12.
- Published Online: 1 MAY 2001
- Published Print: FEB 2000
Culturing by limiting dilution (LD) can be used to estimate the proportion of T cells in a cell preparation which can respond to an activating stimulus to produce interleukin 2 or to generate cytotoxic T lymphocyte (CTL) activity. In the first , IL-2-producing murine T cells are measured following stimulation by the mitogen Con A. This protocol can be modified as described in the first to quantitate responses to alloantigens, anti-CD3 antibody, protein antigens such as keyhole limpet hemocyanin (KLH), and viruses. The second provides a modification for using human responder cells. The second is used for estimating the proportion of cells that can generate a clone of cytotoxic effector cells when stimulated by Con A with the addition of IL-2.