Year: 2023
Type: conference-abstract
Abstract: CD4+ T cells recognize peptide antigens presented on class II Major Histocompatibility Complex (MHC-II) molecules to carry out their function. The remarkable diversity of T cell receptor (TCR) sequences and lack of antigen discovery approaches for MHC-II epitopes has made profiling the specificities of CD4+ T cells challenging. To address this, our lab has expanded our platform of Signaling and Antigen-presenting Bifunctional Receptors (SABRs) to encode MHC-II molecules presenting covalently linked peptides (SABR-IIs) for CD4+ cell antigen discovery. SABR-IIs can present epitopes to CD4+ T cells and induce signaling upon their recognition, generating a readable output. We demonstrate that SABR-IIs libraries presenting endogenous and post-translationally modified epitopes can be used for antigen discovery. Using SABR-II libraries in conjunction with single cell RNA sequencing, we have de-convoluted multiple highly expanded TCRs from pancreatic islets of Non-Obese Diabetic (NOD) mice de novo. We have expanded antigen discovery by incorporating computational TCR similarity prediction metrics followed by experimental validation. Finally, we show that SABR-IIs presenting epitopes in class II HLA alleles can be used for antigen discovery for human CD4+ T cells. Taken together, we have developed a rapid, flexible, scalable, and versatile approach for the de novo identification of CD4+ T cell ligands from single cell RNA sequencing data using experimental and computational approaches. We are continuing to de-convolute the specificity of islet infiltrating CD4+ T cells showing high levels of clonal expansion from both mice and humans with an emphasis on screening theoretical libraries containing hybrid insulin peptides. Supported by grants from NIDDK (1R03DK127447), dkNET New investigator Pilot Program in Bioinformatics, PACER Innovative Discovery Award, JDRF Innovative Immunotherapies Award,
Source: The Journal of Immunology
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W4385687317