Single-Cell Antigen Receptor Sequencing in Pigs with Influenza.

Autor:	Gu W; Division of Animal Sciences, University of Missouri, Columbia, MO, USA.; Bond Life Sciences Center, University of Missouri, Columbia, MO, USA., Madrid DMC; Division of Animal Sciences, University of Missouri, Columbia, MO, USA.; Bond Life Sciences Center, University of Missouri, Columbia, MO, USA., Clements S; Division of Animal Sciences, University of Missouri, Columbia, MO, USA.; Bond Life Sciences Center, University of Missouri, Columbia, MO, USA., Touchard L; Division of Animal Sciences, University of Missouri, Columbia, MO, USA.; Bond Life Sciences Center, University of Missouri, Columbia, MO, USA., Bivins N; Genomics Technology Core, University of Missouri, Columbia, MO, USA., Zane G; Genomics Technology Core, University of Missouri, Columbia, MO, USA., Zhou M; Genomics Technology Core, University of Missouri, Columbia, MO, USA., Lee K; Division of Animal Sciences, University of Missouri, Columbia, MO, USA., Driver JP; Division of Animal Sciences, University of Missouri, Columbia, MO, USA.; Bond Life Sciences Center, University of Missouri, Columbia, MO, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Oct 15. Date of Electronic Publication: 2024 Oct 15.
DOI:	10.1101/2024.10.13.617920
Abstrakt:	Understanding the pulmonary adaptive immune system of pigs is important as respiratory pathogens present a major challenge for swine producers and pigs are increasingly used to model human pulmonary diseases. Single-cell RNA sequencing (scRNAseq) has accelerated the characterization of cellular phenotypes in the pig respiratory tract under both healthy and diseased conditions. However, combining scRNAseq with recovery of paired T cell receptor (TCR) α and β chains as well as B cell receptor (BCR) heavy and light chains to interrogate their repertoires has not to our knowledge been demonstrated for pigs. Here, we developed primers to enrich porcine TCR α and β chains along with BCR κ and λ light chains and IgM, IgA, and IgG heavy chains that are compatible with the 10x Genomics VDJ sequencing protocol. Using these pig-specific assays, we sequenced the T and B cell receptors of cryopreserved lung cells from CD1D -expressing and -deficient pigs after one or two infections with influenza A virus (IAV) to examine whether natural killer T (NKT) cells alter pulmonary TCR and BCR repertoire selection. We also performed paired single-cell RNA and receptor sequencing of FACS-sorted T cells longitudinally sampled from the lungs of IAV-vaccinated and -infected pigs to track clonal expansion in response to IAV exposure. All pigs presented highly diverse repertoires. Pigs re-exposed to influenza antigens from either vaccination or infection exhibited higher numbers of expanded CD4 and CD8 T cell clonotypes with activated phenotypes, suggesting potential IAV reactive T cell populations. Our results demonstrate the utility of high throughput single-cell TCR and BCR sequencing in pigs. Competing Interests: Competing Interest Statement: Authors declare that they have no competing interests.
Databáze:	MEDLINE
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