Unraveling new characteristics of γδ T cells using scRNA-seq in TCR KO chicken

The characterization of T cells in chickens has proven to be challenging, primarily due to the lack of specific markers to differentiate classical Th1, Th2, and Th17 subsets. Furthermore, chickens possess a notably high proportion of γδ T cells, making them a unique model for investigating the poorly understood role of these cells not only in chickens but also in mammals. To gain deeper insights into the functions and characteristics of the αβ and γδ T cell subsets in chickens, whole transcriptome analysis (WTA) on CD3 ⁺ single cells isolated from TCR Cβ knockout (KO), TCR Cγ KO, and wild type (WT) chickens at embryonic day (ED) 18, day (d) 3, and d14 post-hatch was conducted. The results indicate that γδ T cells exhibit cytolytic activity in both TCR Cβ KO and WT chickens, a feature absent in TCR Cγ KO chickens. The severe phenotype observed in TCR Cβ KO chickens revealed a distinct cluster of γδ T cells expressing genes associated with the interferon pathway. Additionally, novel gene markers were identified that more precisely define the αβ and γδ T cell subsets. These findings provide new insights into chicken T cell biology and contribute to a better understanding of the mechanisms underlying the severe phenotype observed in TCR Cβ KO chickens.