UNIT 3.15 Quantitation of Functional T Cells by Limiting Dilution

  1. Richard A. Miller

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0315s35

Current Protocols in Immunology

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.

Author Information

  1. University of Michigan School of Medicine, Ann Arbor, Michigan

Publication History

  1. Published Online: 1 MAY 2001
  2. 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 basic protocol, 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 alternate protocol to quantitate responses to alloantigens, anti-CD3 antibody, protein antigens such as keyhole limpet hemocyanin (KLH), and viruses. The second alternate protocol provides a modification for using human responder cells. The second basic protocol 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.