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Generation of Human Alloantigen-specific T Cells from Peripheral Blood
Published on: November 21, 2014
Generation of human T cell clones
Sabrina Mariotti1, Roberto Nisini
1Dipartimento di Malattie Infettive, Parassitarie e Immunomediate, Istituto Superiore di Sanità, Roma, Italy.
Methods in Molecular Biology (Clifton, N.J.)
|December 3, 2008
Summary
Generating human T cell clones from peripheral blood is challenging due to cell diversity. This guide details methods for T cell cloning, improving experimental clarity and function attribution.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Peripheral blood T lymphocytes exhibit significant heterogeneity, complicating functional attribution in experimental settings.
- A homogeneous population of T lymphocytes (clones) from a single progenitor offers a standardized approach for research.
Purpose of the Study:
- To describe procedures for designing and executing T cell cloning experiments.
- To detail methods for obtaining and maintaining peptide-specific CD4 positive T cell clones from human peripheral blood.
Main Methods:
- Prototypic limiting dilution cloning procedure for isolating peptide-specific CD4+ T cell clones.
- Description of methods for generating diverse T cell clones.
- Brief overview of isolating T cell clones from biopsies and other tissues.
Main Results:
- Detailed protocol for establishing peptide-specific CD4+ T cell clones using limiting dilution.
- Guidance on alternative T cell isolation techniques and sources.
- Inclusion of practical tips to enhance T cell cloning efficiency.
Conclusions:
- Standardized T cell cloning protocols are essential for reproducible immunological research.
- The described methods facilitate the generation of defined T cell populations for functional studies.
- Consideration of alternative cell sources and optimization techniques improves cloning success.
