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Protocol for generation of human peptide-specific primary CD8+ T cell lines
Emma J Grant1, Stephanie Gras1
1Department of Biochemistry and Chemistry, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC 3086, Australia; Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800, Australia.
STAR Protocols
|August 9, 2022
Summary
This protocol expands low-frequency, resting memory peptide-specific CD8+ T cells in vitro. Expanded cells can be characterized using assays like tetramer and intracellular cytokine staining.
Area of Science:
- Immunology
- Cell Biology
Background:
- Peptide-specific CD8+ T cells are crucial for adaptive immunity.
- Directly ex vivo, these cells are infrequent and often quiescent.
Purpose of the Study:
- To detail a protocol for expanding memory peptide-specific CD8+ T cells.
- To enable robust characterization of these expanded cells.
Main Methods:
- In vitro stimulation of peripheral blood mononuclear cells.
- Expansion of peptide-specific CD8+ T cell populations.
- Characterization via tetramer staining and intracellular cytokine staining.
Main Results:
- Successful expansion of memory peptide-specific CD8+ T cells.
- Demonstrated feasibility of subsequent cellular assays.
Conclusions:
- This protocol provides a reliable method for generating sufficient numbers of peptide-specific CD8+ T cells for analysis.
- Facilitates deeper investigation into T cell responses.
