Non-specific cytotoxic cells (NCC) are important cytotoxic leukocytes in teleost immune system. However, the NCC subsets have not been clarified. Thus, we create a comprehensive cell map of ~24,062 head kidney-derived leukocytes from Nile tilapia post poly I:C stimulation using single-cell RNA sequencing (scRNA-seq). Based on cell heterogeneity and known markers, the cells were classified into four cell types including B cell, T cell, NCC and monocytes/macrophages (Mo/MΦ). In the meantime, the regulatory network of NCC population was predicted by WGCNA and four hub genes (Stbd1, VWF, PGP, and GRN) and one transcription factor (Hvcn1) were identified. To further study the differentiation of NCC, four subsets including memory-like NCC, mature NCC, immature NCC, and pre-NCC were revealed in NCC population for the first time. Our data will provide new insight into the biology of NCC and enable more accurate functional and developmental analysis of NCC in immune system of lower vertebrates

非特异性细胞毒性细胞 (NCC) 是硬骨鱼免疫系统中重要的细胞毒性白细胞。但是，NCC 子集尚未阐明。因此，我们使用单细胞 RNA 测序 (scRNA-seq) 在 poly I:C 刺激后创建了来自尼罗罗非鱼的约 24,062 个头肾源性白细胞的综合细胞图谱。根据细胞异质性和已知标志物，将细胞分为四种细胞类型，包括B细胞、T细胞、NCC和单核细胞/巨噬细胞（Mo/MΦ）。同时，通过WGCNA和4个枢纽基因（Stbd1、VWF、PGP和GRN）和一个转录因子（Hvcn1）预测NCC种群的调控网络。) 被识别。为了进一步研究 NCC 的分化，首次在 NCC 人群中揭示了记忆样 NCC、成熟 NCC、未成熟 NCC 和前 NCC 四个亚群。我们的数据将提供对 NCC 生物学的新见解，并能够对低等脊椎动物免疫系统中的 NCC 进行更准确的功能和发育分析