Single-Cell Antigen Receptor Sequencing in Pigs with Influenza

Area of Science:

• Immunology
• Veterinary Medicine
• Genomics

Background:

• Respiratory pathogens pose significant challenges in swine production.
• Pigs serve as valuable models for human pulmonary diseases.
• Characterizing the pig pulmonary adaptive immune system is crucial for understanding disease.

Purpose of the Study:

• To develop and validate primers for high-throughput sequencing of porcine T cell receptor (TCR) and B cell receptor (BCR) repertoires.
• To investigate the impact of natural killer T (NKT) cells on pulmonary TCR and BCR repertoire selection in pigs.
• To track clonal expansion of T cells in response to influenza A virus (IAV) in vaccinated and infected pigs.

Main Methods:

• Development of pig-specific primers for TCR α/β and BCR κ/λ/IgM/IgA/IgG chain enrichment.
• Compatibility testing with 10x Genomics VDJ sequencing protocol.
• Single-cell RNA and receptor sequencing of lung cells and FACS-sorted T cells from IAV-exposed pigs.

Main Results:

• Successful enrichment and sequencing of porcine TCR and BCR repertoires.
• Demonstrated highly diverse TCR and BCR repertoires across all pigs.
• Identified expanded CD4 and CD8 T cell clonotypes with activated phenotypes in pigs re-exposed to IAV antigens.

Conclusions:

• The developed assays enable high-throughput single-cell TCR and BCR sequencing in pigs.
• Re-exposure to IAV through vaccination or infection leads to an increase in expanded, activated T cell clonotypes.
• This work provides a valuable tool for studying adaptive immunity in swine and disease modeling.