Development of an MHC class I Ld-restricted PSA peptide-loaded tetramer for detection of PSA-specific CD8+ T cells in the mouse

Caitlin D Lemke; Jessica B Graham; David M Lubaroff; Aliasger K Salem

doi:10.1038/pcan.2010.57

Back

Development of an MHC class I Ld-restricted PSA peptide-loaded tetramer for detection of PSA-specific CD8+ T cells in the mouse

Journal article

Peer reviewed

Development of an MHC class I Ld-restricted PSA peptide-loaded tetramer for detection of PSA-specific CD8+ T cells in the mouse

Caitlin D Lemke, Jessica B Graham, David M Lubaroff and Aliasger K Salem

Prostate cancer and prostatic diseases, Vol.14(2), pp.118-121

06/2011

DOI: 10.1038/pcan.2010.57

PMCID: PMC3094480

PMID: 21263453

Files and links (1)

url

https://www.ncbi.nlm.nih.gov/pmc/articles/3094480View

Open Access

Abstract

We set out to develop a PSA peptide-loaded tetramer for enumeration of PSA-specific CD8+ T cells in the Balb/c mouse model. A candidate major histocompatibility complex (MHC) class I PSA peptide (HPQKVTKFML188−197) was selected on the basis of its ability to restimulate PSA-specific CD8+ T cells to secrete interferon-γ in our assays. Next, H-2Ld-restricted peptide-loaded and fluorescently labeled tetramers were produced in conjunction with the NIH Tetramer Core Facility, Atlanta, GA, USA. This tetramer was then tested for staining specificity and optimized for detection of PSA-specific CD8+ T cells induced by our PSA-encoding adenovirus tumor vaccine. The MHC class I PSA peptide demonstrated successful restimulation of CD8+ T cells isolated from mice previously vaccinated with a PSA-encoding adenovirus tumor vaccine, with no restimulation observed in control-vaccinated mice. The peptide-loaded H-2Ld tetramer exhibited the desired binding specificity and allowed for detection and frequency determination of PSA-specific CD8+ T cells by flow cytometry. We have successfully designed and validated a PSA peptide tetramer for use in the Balb/c mouse model that can be used to test PSA-based prostate cancer vaccines. Until now, PSA-specific CD8+ T cells in the mouse have only been detectable via cytotoxic T-lymphocyte assays or intracellular cytokine staining, which primarily assess antigen-specific functional activity and not their absolute number. This research tool provides laboratories the ability to directly quantitate CD8+ T cells elicited by PSA-specific immunotherapies and cancer vaccines that are tested in mouse models.

Prostate specific antigen

Intracellular cytokine staining

Mouse

MHC Tetramer

Details

Title: Subtitle: Development of an MHC class I Ld-restricted PSA peptide-loaded tetramer for detection of PSA-specific CD8+ T cells in the mouse
Creators: Caitlin D Lemke - College of Medicine, University of Iowa
Jessica B Graham - College of Medicine, University of Iowa
David M Lubaroff - College of Medicine, University of Iowa
Aliasger K Salem - College of Medicine, University of Iowa
Resource Type: Journal article
Publication Details: Prostate cancer and prostatic diseases, Vol.14(2), pp.118-121
DOI: 10.1038/pcan.2010.57
PMID: 21263453
PMCID: PMC3094480
NLM abbreviation: Prostate Cancer Prostatic Dis
ISSN: 1365-7852
eISSN: 1476-5608
Language: English
Date published: 06/2011
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Urology; Dental Research; Chemical and Biochemical Engineering; Holden Comprehensive Cancer Center
Record Identifier: 9983985716602771

Metrics

28 Record Views

5 Times Cited - Web of Science