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<p>Single-cell RNA sequencing (scRNAseq) has accelerated characterizing cellular phenotypes in pigs under healthy and diseased conditions. To pair scRNAseq with immune receptor profiling, we developed porcine-specific T cell receptor (TCR) and B cell receptor (BCR) enrichment primers that are compatible with the 10 × Genomics VDJ sequencing protocol. Using these assays, we profiled the immune repertoire of cryopreserved lung cells from <i>CD1D</i>-expressing and <i>CD1D</i>-deficient pigs after one or two infections with influenza A virus (IAV) to examine whether natural killer T (NKT) cells influence pulmonary TCR and BCR receptor repertoires. We also profiled T cells longitudinally sampled from the lung fluid of IAV-vaccinated and -infected pigs to track clonal expansion. While all pigs presented highly diverse repertoires, pigs re-exposed to IAV had more expanded T cell clonotypes with activated phenotypes, suggesting potential IAV-reactive clones. Our results demonstrate the utility of high throughput single cell TCR and BCR sequencing in pigs.</p>

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Single-cell antigen receptor sequencing in pigs with influenza

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年7月26日

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  • Weihong Gu,
  • Darling Melany de Carvahlo Madrid,
  • Yuhan Wen,
  • Sadie Clements,
  • Laurie Touchard,
  • Nathan Bivens,
  • Grant Zane,
  • Mingyi Zhou,
  • Kiho Lee,
  • John P. Driver

摘要

Single-cell RNA sequencing (scRNAseq) has accelerated characterizing cellular phenotypes in pigs under healthy and diseased conditions. To pair scRNAseq with immune receptor profiling, we developed porcine-specific T cell receptor (TCR) and B cell receptor (BCR) enrichment primers that are compatible with the 10 × Genomics VDJ sequencing protocol. Using these assays, we profiled the immune repertoire of cryopreserved lung cells from CD1D-expressing and CD1D-deficient pigs after one or two infections with influenza A virus (IAV) to examine whether natural killer T (NKT) cells influence pulmonary TCR and BCR receptor repertoires. We also profiled T cells longitudinally sampled from the lung fluid of IAV-vaccinated and -infected pigs to track clonal expansion. While all pigs presented highly diverse repertoires, pigs re-exposed to IAV had more expanded T cell clonotypes with activated phenotypes, suggesting potential IAV-reactive clones. Our results demonstrate the utility of high throughput single cell TCR and BCR sequencing in pigs.