Fig 1.
Trx-priming attenuates acute allograft rejection and prolong allograft survival.
Orthotopic left lung transplantations from Lewis to Sprague-Dawley rats were performed without immunosuppression using with (+) or without (-) Trx-primed donor lungs. Shown are representative gross images of native lung (right) allograft lung (left) of Trx (-) and Trx (+) priming on day 5 (panels A and B), day 31 (panels G and H), and day 37 (panels K and L) post-transplant, respectively. Shown are representative histological images of Trx (-) and Trx (+) allograft lungs on day 5 (panels C and D), day 7 (panels G and H, and day 31 (panels I and J) post-transplant (original magnification 200x), respectively. Images shown are representative of 3–7 transplants in each group.
Fig 2.
Trx- priming attenuates recipients’ T cell response to the donor type alloantigen accompanied with increased IL-10 production, but not third party control.
Orthotopic left lung transplantations were performed from Lewis to Sprague-Dawley rats without immunosuppression using with (+) or without (-) Trx-primed donor lungs. A: Images of T cell proliferating clusters in the co-culture of 7 day transplant recipients’ CD4+ T cells and naïve donor Lewis DC at day 3 in MLR, image represents Trx (-) and Trx (+) groups respectively. The round dark areas represent T cell clusters. B: 5 day transplant recipients’ spleen CD4+ T cells proliferative response to naive donor DC stimulation in MLR. C: IL-10 concentration in the supernatants of A. D: comparison of 37 day transplanted recipients’ spleen CD4+ T cells proliferative response to the stimulation of donor type antigens from Lewis spleen DC and third party antigens from Wistar spleen DC. E: IL-10 concentration in supernatants of D. Data represent mean ± SE, n = 3. (*p<0.05, **p<0.01 vs Trx (-).
Fig 3.
Trx-priming enhances recipients’ spleen CD4+Foxp3+ regulatory cells.
Panels A, B and C shows representative images of one of three separate flow cytometric analysis of spleen CD4 and Foxp3 intracellular staining on day 5, 21 and 37 post-transplant recipients, respectively. Panel D shows quantitatively analysis of host spleen Foxp3+ CD4+ T cells. Data represent mean ± SE, n = 3.*p<0.05 vs Trx (-).
Fig 4.
Trx-priming inhibits the production of antibodies against donor type antigen.
Two weeks post lung transplant recipient rats were injected with SPC from naïve donor to re-challenge the recipients. The levels of antibodies against donor antigen were measured by immunofluorescent flow cytometry and expressed in MFI. The representative levels of one of three separate host antibodies against donor antigens pre-SPC injection, 1 week, and 2 weeks post-SPC injection are shown in panels A, B, and C, respectively. D: The quantitative levels of host antibodies against donor antigen in pre-SPC, SPC-1 week, and SPC-2 weeks groups (n = 3 for each group). # p< 0.05 vs Trx (-) pre-SPC injection group. *p<0.05, **p<0.01 vs Trx (-) 1 week and 2 weeks, respectively.