Fc-optimized CD40 Agonistic Antibody Induces Tertiary Lymphoid Structures and Systemic Antitumor Immunity in Metastatic Cancer Patients [Human]

Human peripheral blood mononuclear cells (PBMC) from participants of the clinical study were isolated and thawed using prewarmed BD BSA stain buffer (BD Biosciences), washed twice with ice-cold buffer and counted and CD45+ cells were sorted for RNA sequencing. Overall design: Human CD45+ PBMCs isolated by FACS and analyzed by scRNAseq to determine the gene expression profile and T-cell receptor repertoire.

本临床研究受试者的人外周血单个核细胞（peripheral blood mononuclear cells, PBMC）经预热的BD BSA染色缓冲液（BD Biosciences）完成分离与复苏，随后以冰冷缓冲液洗涤两次并进行细胞计数，分选CD45阳性细胞用于RNA测序。整体实验设计：通过荧光激活细胞分选术（fluorescence-activated cell sorting, FACS）分离人CD45+ PBMC，随后采用单细胞RNA测序（single-cell RNA sequencing, scRNAseq）进行分析，以获取基因表达谱与T细胞受体库。