Background Understanding the specific T- and B- cellular immunity during an induced cellular immune response is important for development of anti-tumor immunity in personalized immunotherapy. Advanced single-cell genomics technologies have enabled researchers to do single cells immune profiling, by assessing cell surface proteins, the transcriptome and TCR and/or BCR gene clonotypes. However, understanding antigen-specific recognition at the immune synapse is key to understand the specific immune response in cancer. The dCODE MHC Dextramer® technology combine single-cell genomic profiling with antigen-recognition allowing deep analysis of antigen-specific T-, and B-cells at the single cell level pairing TCR recognition adding to unveil the antigen specific immune response, in cancer and infectious diseases.
Methods We have combined two powerful technologies, Immudex® dCODE Dextramer® (RiO) Reagents and the BD Rhapsody™ Single-Cell Analysis System, to detect and characterize low-frequency antigen-specific T- and B cells, including the full sequences of the V(D)J gene segments of the antigen-specific T- and B cell receptors, as well as profile transcriptome and cell phenotyping by surface protein expression. We used a panel of dCODE Dextramer® (RiO) reagents directed against 10 virus-specific antigens + 6 negative control reagents, spanning MHCI and MHCII, to profile two HPBMC samples, together with 15 BD® AbSeq immune related antibodies, and over 350 immune related genes.
Results We identified 10 different virus-specific T-cell responses over the two samples and revealed major clonotypes of all responses in the two samples, alongside with phenotypic activation profile using the BD® AbSeq reagents and immune target gene expression of the antigen-specific T cells identified by the dCODE® (RiO) Dextramer reagents. Data on antigen-specific B-cell responses is pending.
Conclusions Here we show an assay with the ability to analyze antigen-specific immune cells, by applying combined antigen-specific detection, with surface phenotyping, and gene expression, resulting in a deep phenotypic characterization of the immune cells in the donor. The ability to do high-resolution B and T cell profiling has broader implications and utility in immuno-oncology, infectious diseases, and autoimmunity.
For Research Use Only. Not for use in therapeutic or diagnostic procedures
-Immudex®, Dextramer®, dCODE®, KlickmerTM and U LoadTM are trademarks owned by Immudex ApS
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.