TCR transgenic clone selection guided by immune receptor analysis and single cell RNA expression of polyclonal responders

• DOI: 10.7554/elife.98344.1
• Publication Date: 2024-06-11T15:38:35Z
• Authors: Nincy Debeuf; Sahine Lameire; Manon Vanheerswyn...; Julie Deckers; Caroline De Wolf; Wendy Toussaint; Rein Verbeke; Kevin Verstaen; Hamida Hammad; Stijn Vanhee; Bart N. Lambrecht
• Keywords: Streptamer; clone (Java method)

Since the precursor frequency of naïve T cells is extremely low, investigating early steps antigen-specific cell activation challenging. To overcome this detection problem, adoptive transfer a cohort purified from receptor (TCR) transgenic donors has been extensively used but not readily available for emerging pathogens. Constructing TCR mice hybridomas labor-intensive and sometimes erratic process, since best clones are selected based on antigen-induced CD69 upregulation or IL-2 production in vitro, chains PCR-cloned into expression vectors. Here, we exploited rapid advances single sequencing repertoire analysis to select without hybridoma selection, generated CORSET8 (CORona Spike Epitope specific CD8 cell), carrying protein SARS-CoV-2. Implementing newly created DALI software enabled selection ideal responder clone, antigen reactivity, proliferation immunophenotype vivo. In contrast, traditional method technology was unsuccessful. Identified sequences were inserted as synthetic DNA an vector donor created. After immunization with Spike/CpG-motifs, mRNA vaccination SARS-CoV2 infection, strongly proliferated showed signs activation. Thus, combination scRNA immunophenotyping allowed that can be generate mice.