Fig. 1.
Primed macrophages rejected allogeneic donor splenocytes and skin grafts with memory. a The flowchart for the donor and third-party splenocyte rejection assays in SCID mice with adoptive transfer of primed macrophages is shown. BALB/c mice were immunized with B6 splenocytes. By 10 or 120 days after immunization, the sorted B6 splenocyte-priming BALB/c PEMs and CFSE-labeled target splenocytes (BALB/c cells: B6/FvB cells = 1:1) were mixed at a 4:1 ratio and intraperitoneally injected into SCID mice. The survival of CFSE-labeled cells (b) and the total percentages of CFSEhigh cells among total CFSE-labeled cells (c) in SCID mice were determined 24 h later by flow cytometry. d The experimental procedure for investigation of donor and third-party skin graft rejection in SCID mice with adoptive transfer of primed macrophages is shown. BALB/c mice were immunized with B6 splenocytes. By 10 or 120 days after immunization, the sorted naive or primed BALB/c PEMs were adoptively transferred into MHC-matched SCID mice by i.v. injection, followed by transplantation of allogeneic priming donor B6 or third-party allogeneic FvB skin grafts. e The survival of B6 and FvB skin grafts in the SCID mice is shown. f The number of days of survival of allogeneic skin grafts in the SCID mice are summarized. g The percentages of transferred BALB/c splenocyte-primed B6-GFP+ macrophages were detected in the peripheral blood, peritoneal cavity, and spleen in syngeneic Rag2 KO mice 130 days after adoptive transfer. **P < 0.01 compared with the indicated groups