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Published on: June 4, 2019
Quantitation of functional T cells by limiting dilution
1University of Michigan School of Medicine, Ann Arbor, Michigan, USA.
Current Protocols in Immunology
|April 25, 2008
Summary
Limiting dilution assays quantify T-cell responses, measuring interleukin-2 production or cytotoxic T-lymphocyte activity. These methods enable precise estimation of responsive T-cell proportions in various immune cell preparations.
Area of Science:
- Immunology
- Cell Biology
Background:
- Limiting dilution (LD) is a technique used to estimate the frequency of cells with specific functional properties.
- T cells play a crucial role in adaptive immunity, mediating both helper and cytotoxic functions.
Purpose of the Study:
- To describe protocols for quantifying T-cell responses using limiting dilution.
- To detail methods for measuring interleukin-2 (IL-2) production and cytotoxic T-lymphocyte (CTL) activity.
Main Methods:
- The study outlines two basic protocols and modifications for limiting dilution assays.
- Protocol 1 measures IL-2-producing murine T cells stimulated by Concanavalin A (Con A), with variations for alloantigens, anti-CD3 antibody, protein antigens (KLH), and viruses.
- Protocol 2 estimates the proportion of cells generating CTL activity upon stimulation with Con A and IL-2.
Main Results:
- The described limiting dilution methods allow for the quantitative estimation of T cells responding to various stimuli.
- Modifications enable the use of human cells and the assessment of cytotoxic effector cell generation.
Conclusions:
- Limiting dilution assays provide a robust method for determining the frequency of functional T cells.
- These protocols are adaptable for diverse research applications in immunology and cell biology.
