Reconstitution of Immune System in Mice

On the day of inoculation with tumor segments, human PBMCs were prepared according to a previous study (19 (link)). Fresh peripheral venous blood from healthy adult volunteers was collected at The First Affiliated Hospital of Guangzhou Medical College (Guangzhou, China) in heparinized tubes. For the isolation of PBMCs, leucosep tubes (Greiner Bio-One, Wemmel, Belgium) were used, and blood was diluted 1:1 with RPMI-1640 medium (vol/vol) prior to transferring into the leucosep tube. Following centrifugation (10 min, 1000 × g), the PBMC layer was pooled and transferred into a 15-ml falcon tube. The sample was washed with 10 ml phosphate-buffered saline (PBS) and centrifuged again for 10 min at 250 × g. The obtained cell pellet was resuspended in PBS. A total of 1×10⁸ PBMCs per mouse were intraperitoneally injected into NOD/SCID mice of the Am1010+PBMC and PBMC groups, for the reconstitution of immune system. The same volume of PBS was injected into the mice of the Am1010 group.