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Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
Quantifying viable virus-specific T cells without a priori knowledge of fine epitope specificity
Carol Beadling1, Mark K Slifka
1Vaccine and Gene Therapy Institute, Oregon Health & Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
Nature Medicine
|October 3, 2006
Summary
A new assay, T-cell recognition of APCs by protein transfer (TRAP), detects pathogen-specific T cells. This method identifies T cells by tracking protein transfer from antigen-presenting cells, independent of specific epitopes.
Area of Science:
- Immunology
- Cell Biology
Background:
- Synthetic peptide-MHC tetramers aid pathogen-specific T cell identification.
- Undefined epitopes limit current T cell detection methods, requiring epitope-independent approaches.
Purpose of the Study:
- To develop a novel assay for detecting T cells that functions independently of specific peptide-MHC recognition.
- To leverage the phenomenon of lymphocytes acquiring surface proteins from antigen-presenting cells (APCs).
Main Methods:
- Developed the T-cell recognition of APCs by protein transfer (TRAP) assay.
- Employed biotinylation and streptavidin-fluorochrome labeling of APCs.
- Monitored the acquisition of labeled APC proteins by T cells.
Main Results:
- The TRAP assay detects MHC class I-restricted T cells.
- The method is independent of T cell cytokine profiles and peptide-MHC affinities.
- TRAP provides a versatile tool for monitoring APC membrane acquisition by antigen-specific T cells.
Conclusions:
- The TRAP assay offers a novel, epitope-independent method for identifying antigen-specific T cells.
- This technique facilitates the study of T cell interactions with APCs through protein transfer.
