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:

 

 

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, 1996 Oct 4;274(5284):94-6.

Scientific Papers using immunAware reagents

Oberhardt, V., et al., Rapid and stable mobilization of CD8(+) T cells by SARS-CoV-2 mRNA vaccine. Nature, 2021.

Sahin, U., et al., BNT162b2 vaccine induces neutralizing antibodies and poly-specific T cells in humans. Nature, 2021.

Tippalagama, R., et al., HLA-DR Marks Recently Divided Antigen-Specific Effector CD4 T Cells in Active Tuberculosis Patients. J Immunol, 2021.

Bigot, J., et al., Splicing patterns in SF3B1 mutated uveal melanoma generate shared immunogenic tumor-specific neo-epitopes. Cancer Discov, 2021.

Lande, R. et al. Complementary Effects of Carbamylated and Citrullinated LL37 in Autoimmunity and Inflammation in Systemic Lupus Erythematosus. International Journal of Molecular Sciences 22, 1650 (2021).

Schulien, I. et al. Characterization of pre-existing and induced SARS-CoV-2-specific CD8+ T cells. Nature Medicine 27, 78-85, doi:10.1038/s41591-020-01143-2 (2021).

Osterbye T, Nielsen M, Dudek NL, Ramarathinam SH, Purcell AW, Schafer-Nielsen C, et al. HLA Class II Specificity Assessed by High-Density Peptide Microarray Interactions. J Immunol. 2020;205(1):290-9.

Mohamed YS, Borthwick NJ, Moyo N, Murakoshi H, Akahoshi T, Siliquini F, et al. Specificity of CD8(+) T-Cell Responses Following Vaccination with Conserved Regions of HIV-1 in Nairobi, Kenya. Vaccines (Basel). 2020;8(2).

Lande R, Palazzo R, Gestermann N, Jandus C, Falchi M, Spadaro F, et al. Native/citrullinated LL37-specific T-cells help autoantibody production in Systemic Lupus Erythematosus. Sci Rep. 2020;10(1):5851.

Acevedo GR, Juiz NA, Ziblat A, Perez Perri L, Girard MC, Ossowski MS, et al. In Silico Guided Discovery of Novel Class I and II Trypanosoma cruzi Epitopes Recognized by T Cells from Chagas’ Disease Patients. J Immunol. 2020

Ndhlovu ZM, Kazer SW, Nkosi T, Ogunshola F, Muema DM, Anmole G, et al. Augmentation of HIV-specific T cell function by immediate treatment of hyperacute HIV-1 infection. Sci Transl Med. 2019;11(493).

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.

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.

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).

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.

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.

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.

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.