MHC Tetramers are used for the detection of antigen-specific T-cell populations. MHC-peptide tetramer complexes recognize antigen specific T-cells at the single cell level, allowing for accurate measurements of the functional immune response in cancer, infectious diseases, and autoimmune diseases. Due to their increased avidity, MHC tetramers stably bind to cognate TcRs and are evaluated using flow cytometry.
T lymphocytes play a central role in immune system. Total Tcell and T cell subset counts are measured by detection of various cellsurface molecules. Enumeration of CD4+antigen-specific T cells requirescognate recognition of the T cell receptor (TCR) by a class II MHC/peptidecomplex. This can be done using T-SelectMHC Class II Tetramers which arecomposed of four MHC class II molecules each bound to the specificpeptide1, 2and conjugated with a fluorescent protein. Thus, T-SelectMHCTetramer assaysallow quantitation of thetotalT cell population specific for agiven peptide complexed in a particular MHC molecule. Furthermore, sincebinding does not depend on functional pathways, Tetramer-stainedpopulation includesspecific CD4+T cells regardless of functional status. Measurementsmay be performed in whole blood or isolated lymphocyte/mononuclearcell preparations.In some cases where frequency is low, it may benecessary to perform an in vitrocell expansion3. Specific cell stainingis accomplished by incubating the sample with the T-SelectMHC Tetramerreagent, then washing away excess Tetramer. The number of Tetramerpositive lymphocytes is then determined by flow cytometry.
This Tetramer reagent comprises murineclass II MHC I-Aband epitope peptide derived from ovalbumin (OVA), anditcan detect an I-Ab-restrictedOVA323-339-specific CD4+T cells.The CD4+T cell epitope,OVA323-339, of the OVA model antigen hasbeen a useful tool in immunology. The OT-II transgenic mouse strain carriesa TCR transgenespecific for the OVA323-339peptide,and it is useful for studyingT cell immunology.
A Tetramer, which is constructed with the same allele(I-Ab)of interest and an irrelevant peptide, may be used as a negative controlTetramer. Alternatively, a cell population devoid of Tetramer-positive cells may be used as a negative control.
Product Specifications
Application
FC
Reactivity
Mouse
Conjugation
PE
Immunogen
OVA
Peptide Sequence
ISQAVHAAHAEINEAGR
Class
Class II
Sequence Location
323-339
MHC Allele
I-Ab
Documents & Links for T-Select I-Ab OVA 323-339 Tetramer-PE
