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. 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（human peripheral blood mononuclear cells, PBMC）采用预热的BD BSA染色缓冲液（BD Biosciences）完成分离与复苏解冻，随后以预冷缓冲液洗涤两次并进行细胞计数，分选出CD45阳性细胞用于RNA测序。通过荧光激活细胞分选术（fluorescence-activated cell sorting, FACS）分离得到的人CD45阳性PBMC，经单细胞RNA测序（single-cell RNA sequencing, scRNAseq）分析以明确其基因表达谱与T细胞受体库。