bims-hummad 2025-10-19 papers

Curr Protoc. 2025 Oct;5(10): e70224

Humanized Mouse Models for Type 1 Diabetes.

David V Serreze, Marissa Tousey-Pfarrer, Jeremy J Racine.

T cell-mediated autoimmune type 1 diabetes (T1D) is under complex polygenic control in both humans and the NOD mouse model. However, in both species, particular major histocompatibility complex (MHC; designated HLA in humans) haplotypes provide the primary T1D risk factor. Both MHC/HLA class I and II variants interactively contribute to T1D by respectively driving autoreactive CD8 and CD4 T cell responses that cooperatively destroy insulin-producing pancreatic β cells. While NOD mice have provided important insights to the pathogenic basis of T1D, the model has so far provided only a limited means to identify possible clinically translatable disease intervention approaches. This highlights a need to humanize NOD mice in ways that their pathogenic basis of T1D development becomes more similar to that characterizing the disease course in patients. In this review, we discuss the use of CRISPR/Cas9-generated murine-MHC-deficient NOD mice as a platform for introduction of patient-relevant HLA and T cell receptor molecules. These mice provide ever-improving models for development of clinically applicable interventions for T1D and other autoimmune diseases. © 2025 The Author(s) Current Protocols published by Wiley Periodicals LLC.

Keywords: MHC/HLA; TCRs; autoimmune T1D; humanized mice

DOI: https://doi.org/10.1002/cpz1.70224

Nat Commun. 2025 Oct 13. 16(1): 9090

HLA matching or CRISPR editing of HLA class I/II enables engraftment and effective function of allogeneic human regulatory T cell therapy in a humanized mouse transplantation model.

Regulatory T cells (Tregs) hold promise for treating autoimmune disease and transplant rejection, yet generation of autologous products for adoptive transfer can suffer donor variability and slow turnaround, limiting their use in urgent indications. We therefore examine whether allogeneic, pre-manufactured ('off-the-shelf') Tregs could overcome these barriers. In a human skin-xenograft model, HLA-mismatched Tregs are swiftly eliminated by recipient CD8+ T cells and fail to protect grafts. Stringent matching of HLA class I and II restores efficacy but is clinically impractical. Using non-viral CRISPR editing we disrupt B2M and CIITA while inserting an HLA-E-B2M fusion, generating hypo-immunogenic Tregs that evade both T and NK cell attack. Engineered cells retain FOXP3 stability and potent in vitro suppression, and after a single low-dose infusion, prolong human skin graft survival in a humanized mouse model comparably to autologous Tregs. Histology and spatial transcriptomics reveal minimal cytotoxic infiltration and enrichment of immunoregulatory and tissue-repair programmes. Multiplex HLA engineering thus enables ready-to-use allogeneic Tregs that withstand host immune attack for adoptive transfer.

DOI: https://doi.org/10.1038/s41467-025-64945-3

Curr Protoc. 2025 Oct;5(10): e70225

Autoantigen-Humanized Mouse Models of Bullous Pemphigoid.

Takuya Kawamura, Hideyuki Ujiie.

Bullous pemphigoid (BP) is an autoimmune blistering disease characterized by tense blisters and itchy erythema, predominantly affecting elderly individuals. The pathogenic autoantibodies mainly target collagen XVII (COL17), leading to subepidermal blister formation and infiltrations of immune cells, including eosinophils and neutrophils. Although systemic oral corticosteroids remain the mainstay of treatment, their use in elderly patients is often limited by serious complications and adverse effects, highlighting the unmet need for novel therapeutic targets. Several mouse models for BP have been reported; however, the inconsistency of disease induction has hindered therapeutic investigations. The active BP mouse model has emerged as a reliable system that recapitulates key disease features, making it valuable for both preclinical therapeutic studies and elucidation of BP pathophysiology. Furthermore, autoantigen-humanized mouse models, including the active BP model and the neonatal passive IgG transfer model, provide significant advantages for the development of antigen-specific therapies. Here, we describe detailed materials and methods for mouse models for BP using the COL17-humanized mouse, including the active BP mouse model and the neonatal passive IgG transfer model. Protocol modifications may be necessary when using different donor mice and antibodies. © 2025 Wiley Periodicals LLC. Basic Protocol 1: Active BP mouse model Basic Protocol 2: Passive IgG transfer mouse model using neonatal mice.

Keywords: adoptive transfer; autoantigen‐humanized mouse; bullous pemphigoid; passive IgG transfer; skin graft

DOI: https://doi.org/10.1002/cpz1.70225