PSGL-1-mTOR-TIM3 Signaling in a Murine Model of Hindlimb Transplantation
1UCLA Medical Center, Los Angeles, CA, 2Johns Hopkins UJohns Hopkins University School of Medicine, Baltimore, MD
Meeting: 2022 American Transplant Congress
Abstract number: 960
Keywords: Graft survival, Immunosuppression, T cell activation
Topic: Basic & Clinical Science » Basic & Clinical Science » 20 - VCA
Session Information
Session Time: 7:00pm-8:00pm
Presentation Time: 7:00pm-8:00pm
Location: Hynes Halls C & D
*Purpose: Peri-transplant tissue damage driven by innate-adaptive immune cascades resulting from ischemia-reperfusion injury (IRI) is a major concern in vascularized composite allotransplantation (VCA). Here, we targeted the endothelial CD62 – T-cell CD162 signaling pathway with a recombinant soluble form of a tandem-PSGL-1 (TSGL-Ig) to monitor organ preservation-triggered early/late immune responses in VCAs.
*Methods: Orthotopic hindlimb transplantation studies were performed to determine the significance of CD62-CD162 signaling in the context of innate/adaptive immune activation in syngeneic (BL6->BL6) vs. allogeneic (Balb/c->BL6) mouse models with/without TSGL-Ig during 24h cold-storage (UW). To determine whether TSGL-Ig promotes CD4+ T cell exhaustion, allo-VCAs pre-conditioned with TSGL-Ig, were treated with adjunctive anti-TIM3 Ab. Some TSGL-Ig treated allo-VCAs were conditioned with Rapamycin (Rapa), an mTOR inhibitor, to determine whether targeting endothelial CD62 – T-cell CD162 by TSGL-Ig synergizes with immunosuppressive Rapa. Skin and muscle samples were collected and analyzed at 24h post-transplant.
*Results: Treatment with TSGL-Ig exerted anti-inflammatory function in the skin and inhibited thrombosis in the muscle of syn-VCAs. By contrast, TSGL-Ig treated allo-VCAs showed a more remarkable improvement, evidenced by decreased H&E scores (p<0.05), and suppressed T-cell activation markers (e.g. IFNg, T-Bet and CD4/8), as detected by qPCR. TSGL-Ig treatment also led to higher TIM-3 gene expression while mTOR signaling was down-regulated. Anti-TIM3 Ab used as an adjunctive therapy increased tissue damage and T-cell activation, as compared with TSGL-Ig monotherapy, suggesting the beneficial effect of TSGL-Ig may relate to TIM-3 - mTOR axis. Interestingly and by contrast, TSGL-Ig failed to attenuate VCA-induced tissue damage with unchanged TIM-3 levels in allo-VCA groups conditioned with Rapa, suggesting TSGL-Ig improved VCA was mTOR-TIM-3 signaling dependent. Skin samples from Rapa + TSGL-Ig regimen showed increased T-cell activation and inferior long-term VCA survival. Lastly, TSGL-Ig failed to up-regulate TIM-3 in syn-VCA, suggesting TIM-3 signaling is essential in an adaptive immune response in allo-VCA.
*Conclusions: Our results identify the PSGL-1-TIM-3 signaling axis as a novel regulator of adaptive immunity in VCA, with mTOR being the critical checkpoint of this pathway.
To cite this abstract in AMA style:
Kadono K, Messner F, He J, Guo Y, Oh B, Brandacher G, Kupiec-Weglinski J. PSGL-1-mTOR-TIM3 Signaling in a Murine Model of Hindlimb Transplantation [abstract]. Am J Transplant. 2022; 22 (suppl 3). https://atcmeetingabstracts.com/abstract/psgl-1-mtor-tim3-signaling-in-a-murine-model-of-hindlimb-transplantation/. Accessed December 2, 2024.« Back to 2022 American Transplant Congress