Mouse Models and Experimental Protocols to Study Alloantibody-Mediated Transplant Rejection.

Abstract

Transplantation is the definitive treatment for patients with end-stage organ failure. Following allogeneic transplant, the recipient's immune system recognizes transplanted cells or organs as foreign. The immune system recognizes and targets the foreign tissue for damage through cell-mediated rejection (CMR) and/or antibody-mediated rejection (AMR). Immunosuppressive agents are utilized to protect the transplant from rejection and extend transplant function and survival. Despite advances in immunosuppressive agents, AMR remains a critical barrier to the success of transplantation. AMR occurs when B cells produce alloantibodies that bind the allograft causing antibody-dependent, complement-mediated or immune cell-mediated cytotoxic damage. Continued research on AMR is required to develop novel and effective therapeutic strategies. Murine AMR models have been utilized to investigate mechanisms mediating the production of posttransplant alloantibodies and the pathology of damage to the transplanted allograft. These models facilitating the investigation of cellular and molecular mechanisms of alloantibody production and allograft damage are critical to the development of novel therapeutic strategies to prevent and treat AMR. This article describes the methodologies used to study AMR in animal transplant models. These include protocols to detect and measure alloantibodies, allograft survival, AMR pathology, and effector immune cell responses following transplantation. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Alloserum transfer into immune-incompetent recipient mice to determine transplant organ susceptibility to AMR Basic Protocol 2: Allogeneic transplantation into immune-deficient mice to study critical cellular and molecular pathways impacting AMR Support Protocol 1: Quantification of posttransplant alloantibody titer Support Protocol 2: Monitoring of transplant allograft survival Support Protocol 3: Assessment of immunopathology and severity of AMR Basic Protocol 3: Analysis of posttransplant immunologic responses.