Multidimensional Analysis of Immune Rejection in a Murine HLA ‐Mismatched Laryngeal Transplant Model

Objective To establish a murine heterotopic laryngeal transplantation (LT) model with a single HLA‐A2 mismatch and to develop a multidimensional framework for dynamic evaluation of immune rejection and graft injury. Methods Laryngeal grafts from HLA‐A2 transgenic mice were transplanted into C57BL/6 recipients to generate an HLA‐mismatched LT model, with syngeneic C57BL/6 → C57BL/6 transplants as controls. A multidimensional evaluation system was established, integrating physiological monitoring (survival, body weight, graft appearance, neck circumference, computed tomography, and ultrasonography), immunological profiling (histopathology, immunohistochemistry, and flow cytometry), and inflammatory assessments (cytokines and biochemical parameters) from Day 0 to 12 weeks posttransplantation. Results Compared with syngeneic transplantation, the HLA‐mismatched group showed aggravated immune rejection and graft injury. Progressive graft volume reduction was an early discriminator, detectable at 1 week posttransplantation and correlating with graft surface darkening and ulceration. Immunophenotyping revealed early CD4 + and CD8 + T‐cell infiltration beginning at 1 week, whereas regulatory T cells increased at 3 days before markedly declining. Cytokine profiling showed elevations of MIP‐1β, G‐CSF, and IL‐10 from 4 weeks onward, consistent with immune activation and graft injury. Conclusion A murine heterotopic LT model with an HLA‐A2 mismatch was established, demonstrating that a single HLA Class I disparity was sufficient to drive progressive immune rejection and graft injury in LT. A panel of biomarkers associated with adverse graft outcomes was observed, providing indicators for monitoring HLA‐driven immune injury. Level of Evidence N/A.