MHC Monomers and Tetramers

RESOURCES

PROTOCOLS

A tetramer assay (tetramer stain) is used to detect and quantify antigen specific T cells. The interaction between a peptide MHC complex (pMHC) and a cognate T cell receptor (TcR) is of too low affinity to be detected in flow cytometry. Multimerization of monomeric biotinylated pMHC by streptavidin (1) complexing increases the avidity of the pMHC streptavidin tetramer towards cognate TcRs to allow detection of antigen specific T cells. immunAware has three product lines for MHC class I:

ready-to-use MHC tetramers loaded with peptide from our catalog
custom made monomers or tetramers based on your choice of peptide-MHC combination
Make you own tetramers with our peptide receptive MHC molecules easYmer®

We have developed dedicated staining protocols for CD4+ and CD8+ T cells to ensure highly specific staining with our Tetramers. For optimal results, please follow our MHC peptide loading protocol and tetramer staining protocols developed to obtain high peptide-MHC complex yields with the easYmer kits and subsequently stain T cells.

Read more about easYmers and Tetramers

References

(1) Altman JD. et al, Science. 1996 Oct 4;274(5284):94-6.

easYmer Protocols

Assessment of HLA-complex folding – FACS assay

easYmer – Tetramer production protocol

Calculate amount of Streptavidin-conjugate

Tetramer Staining protocols

HLA class I tetramer staining of human T cells

HLA class II tetramer staining of human T cells

Scientific Papers

1. Moyo N, Vogel AB, Buus S, Erbar S, Wee EG, Sahin U, et al. Efficient Induction of T Cells against Conserved HIV-1 Regions by Mosaic Vaccines Delivered as Self-Amplifying mRNA. Mol Ther Methods Clin Dev. 2019;12:32-46.

2. Gonzalez-Duque S, Azoury ME, Colli ML, Afonso G, Turatsinze JV, Nigi L, et al. Conventional and Neo-antigenic Peptides Presented by beta Cells Are Targeted by Circulating Naive CD8+ T Cells in Type 1 Diabetic and Healthy Donors. Cell Metab. 2018;28(6):946-60 e6.

3. Culina S, Lalanne AI, Afonso G, Cerosaletti K, Pinto S, Sebastiani G, et al. Islet-reactive CD8(+) T cell frequencies in the pancreas, but not in blood, distinguish type 1 diabetic patients from healthy donors. Sci Immunol. 2018;3(20).

4. Nastasi C, Fredholm S, Willerslev-Olsen A, Hansen M, Bonefeld CM, Geisler C, et al. Butyrate and propionate inhibit antigen-specific CD8(+) T cell activation by suppressing IL-12 production by antigen-presenting cells. Sci Rep. 2017;7(1):14516.

5. Abana CO, Pilkinton MA, Gaudieri S, Chopra A, McDonnell WJ, Wanjalla C, et al. Cytomegalovirus (CMV) Epitope-Specific CD4(+) T Cells Are Inflated in HIV(+) CMV(+) Subjects. J Immunol. 2017;199(9):3187-201.

6. Svitek N, Hansen AM, Steinaa L, Saya R, Awino E, Nielsen M, et al. Use of “one-pot, mix-and-read” peptide-MHC class I tetramers and predictive algorithms to improve detection of cytotoxic T lymphocyte responses in cattle. Vet Res. 2014;45:50.

7. Leisner C, Loeth N, Lamberth K, Justesen S, Sylvester-Hvid C, Schmidt EG, et al. One-pot, mix-and-read peptide-MHC tetramers. PLoS One. 2008;3(2):e1678.