Anti-CTLA-4 Treatment Abrogates Co-stimulation Blockade-induced Acceptance of Transgenic Porcine Islets in Humanized Mice

Transplantation. 2025 Dec 1;109(12):e691-e696. doi: 10.1097/TP.0000000000005512. Epub 2025 Aug 26.

Abstract

Background: In previous studies, we showed that beta cell-specific overexpression of high-affinity variant of human CTLA-4 (LEA29Y), a high-affinity variant of cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4)-immunoglobulin, prevented porcine islet rejection in humanized mouse models. We here investigate whether long-term xenograft function and survival is maintained after neutralization of LEA29Y-mediated co-stimulation blockade.

Methods: Diabetic humanized NOD-SCID IL2rγ -/- mice were transplanted with transgenic neonatal porcine islet-like clusters expressing LEA29Y under control of the porcine insulin promoter. After development of normal glucose tolerance, mice were treated with blocking anti-CTLA-4 antibody (Ab) or isotype control Ab. Reoccurrence of diabetes, plasma cytokines/chemokines and graft histology were analyzed.

Results: Systemic treatment with an inhibitory anti-humanized CTLA-4 Ab led to a significant increase of pro-inflammatory plasma cytokine production (interferon gamma, monokine induced by interferon gamma; P < 0.05) at day 14 and reoccurrence of diabetes in 100% of the animals within 40 d after Ab application ( P = 0.01). Strong infiltration with human CD45 + cells consisting mainly of CD4 + and CD8 + T cells and some B lymphocytes and few destructed remaining beta cells were observed in the treatment group indicating rapid and severe graft rejection.

Conclusions: The present data demonstrate that long-lasting acceptance of LEA29Y transgenic porcine islet grafts is dependent on CTLA-4/LEA29Y signaling even several months after transplantation. This finding has important implications on safety of pig islet xenotransplantation in patients with type 1 diabetes because it provides a potent tool for graft elimination in case of medical indication.

MeSH terms

  • Animals
  • CTLA-4 Antigen* / antagonists & inhibitors
  • CTLA-4 Antigen* / genetics
  • CTLA-4 Antigen* / immunology
  • CTLA-4 Antigen* / metabolism
  • Cytokines / blood
  • Diabetes Mellitus, Experimental* / surgery
  • Graft Rejection* / immunology
  • Graft Rejection* / prevention & control
  • Graft Survival* / drug effects
  • Humans
  • Interleukin Receptor Common gamma Subunit / deficiency
  • Interleukin Receptor Common gamma Subunit / genetics
  • Islets of Langerhans Transplantation* / immunology
  • Islets of Langerhans Transplantation* / methods
  • Islets of Langerhans* / drug effects
  • Islets of Langerhans* / immunology
  • Islets of Langerhans* / metabolism
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Mice, Transgenic
  • Swine
  • Time Factors
  • Transplantation, Heterologous

Substances

  • CTLA-4 Antigen
  • Cytokines
  • Interleukin Receptor Common gamma Subunit
  • CTLA4 protein, human
  • Il2rg protein, mouse