Conserved functional features of natural killer cell subsets in chicken, human, and murine immune systems

Immunological conservation across species is crucial for comparative biology. While Natural killer (NK) cells' functions are conserved in humans and mice at the single-cell level, they remain unexplored in chickens due to technical limitations. Single-cell RNA sequencing in a recombination-activating gene 1-deficient (RAG1 ^-/-) chicken model revealed two NK cell subpopulations (NK-1 and -2) in chickens, analogous to those in humans and mice. Cross-species analysis revealed that most genes exhibited distinct expression patterns within NK subsets, reflecting evolutionary divergence, though functional genes were conserved across chicken, humans, and mice. NK-1 cells exhibited conserved cytotoxic functions through immunological synapses and activated signaling pathways, while NK-2 cells exhibited conserved immune-regulatory functions via cytokine production. Transcription factors related to NK cells' terminal and early maturation were upregulated in NK-1 and NK-2 cells, respectively. These findings highlight evolutionarily conserved immune mechanisms, establishing chickens as potential avian models for translational research in developing treatments against infectious diseases.