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

• DOI: 10.7554/elife.98344.3
• Publication Date: 2024-12-30T15:30:41Z
• 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: clone (Java method); Streptamer

Since the precursor frequency of naive 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 polymerase chain reaction (PCR)-cloned into expression vectors. Here, we exploited rapid advances single-cell sequencing repertoire analysis to select without hybridoma selection, generated CORSET8 ( COR ona S pike E pitope specific CD8 cell), carrying Spike protein SARS-CoV-2. Implementing newly created DALI software enabled selection ideal responder clone, antigen reactivity, proliferation, immunophenotype vivo. Identified sequences were inserted as synthetic DNA an vector donor created. After immunization with Spike/CpG-motifs, mRNA vaccination SARS-CoV-2 infection, strongly proliferated showed signs activation. Thus, combination scRNA immunophenotyping allowed that can be generate mice.