scRNA-seq of human CD45+ cells proliferated in the salivary grand or peripheral blood of CD8T cells depleted PBMC-transferred humanized mice

We found that the transfer of PBMCs depleted of CD8 T cells into immunodeficient mice resulted in the development of Sjögren's syndrome-like salivary gland symptoms without causing GVHD. To characterize the human immune cells infiltrating the salivary glands, we examined the profiles of human immune cells in the salivary glands or peripheral blood of humanized mice by 10x Genomics single-cell RNA sequencing (scRNA-seq). We revealed that human Tph-like cells co-expressing IL-21 and CXCL13 were highly infiltrated in the salivary glands of CD8 T cell-depleted humanized mice. Overall design: Peripheral blood or salivary glands were collected from immunodeficient mice transplanted with either human PBMCs or PBMCs depleted of CD8 T cells, and mononuclear cells were isolated by density gradient centrifugation. Human CD45+ cells were sorted by fluorescence-activated cell sorting (FACS) using a BD Aria III, and scRNA-seq analysis was performed.

我们发现，将去除CD8 T细胞（CD8 T cells）的外周血单个核细胞（PBMC）移植到免疫缺陷小鼠（immunodeficient mice）体内后，可诱发类干燥综合征（Sjögren's Syndrome）样唾液腺症状，且不会引发移植物抗宿主病（GVHD）。为表征浸润唾液腺的人类免疫细胞，我们通过10×基因组学（10x Genomics）单细胞RNA测序（scRNA-seq）分析了人源化小鼠（humanized mice）唾液腺及外周血中的人类免疫细胞谱。我们发现，共表达白细胞介素21（IL-21）与趋化因子C-X-C基序配体13（CXCL13）的人类外周辅助T细胞样（Tph-like）细胞在去除CD8 T细胞的人源化小鼠唾液腺中呈现高度浸润特征。实验整体设计：分别从移植了完整人类外周血单个核细胞或去除CD8 T细胞的外周血单个核细胞的免疫缺陷小鼠体内采集外周血与唾液腺样本，通过密度梯度离心法分离单个核细胞；采用BD Aria III流式细胞仪通过荧光激活细胞分选（FACS）筛选人类CD45阳性细胞，并开展单细胞RNA测序分析。