Development of a humanized mouse model to analyze antibodies specific for human leukocyte antigen (HLA)

Abstract

In organ transplantation, human leukocyte antigen (HLA)-mismatch grafts not only induce the activation of cellular mediated immune response but also the development of chronic antibody-mediated rejection due to the donor-specific anti-HLA antibody (DSA) produced by B cells and plasma cells interacting with the graft endothelium. Significant improvement in long-term survival after transplantation can be expected if antibody-mediated rejection due to the DSA can be overcome. However, the mechanism of producing or controlling the DSA remains to be elucidated. In recent decades, “humanized” mouse models have been widely used for the basic research of human immune systems, but a humanized mouse model to analyze the mechanism of DSA production has not been established yet. Thus, we aimed to create a humanized mouse using a severe immunodeficiency mouse (NSG mouse) administered with human peripheral blood mononuclear cells (PBMCs). Initially, we detected a very low level of human total-IgG and no anti-HLA antibodies (Abs) in these mice. In our next attempt, we mixed PBMCs of various HLA antigenic combinations with or without regulatory T cells and preconditioned them by culturing on feeder cells stably transfected with human CD40 ligand (h-CD40L) alone or with h-CD40L and human B cell activating factor (h-BAFF). They were subsequently co-cultured with the corresponding irradiated stimulator PBMCs, and all cells were administered into naïve NSG mice. Although all three humanized models had sufficient human total-IgG and anti-HLA antibody production, allospecific anti-HLA Ab production was prominently suppressed whereas non-specific anti-HLA Abs were sufficiently detected. Therefore, this novel humanized mouse model might be useful for analyzing the mechanism of anti-allogeneic human B cell tolerance induction.

A novel cDNA‐uPA/SCID/Rag2−/−/Jak3−/− mouse model for hepatitis virus infection and reconstruction of human immune system

“…human immune system chimeric mouse model without humanized liver 27. In our mouse model, we confirmed immune cell accumulation, especially in human hepatocyte areas and detection of transplanted human hepatocyte-specific anti-HLA antibody.Antibody-mediated allograft rejection due to donor-specific anti-HLA antibody influences transplantation outcomes despite advances in immunosuppression and antibody screening prior to F I G U R E 4 PBMC administration into human hepatocyte-transplanted cDNA-uPA/SCID/Rag2 −/− /Jak3 −/− mice.…”

“…However, no alloimmunity mouse model was reported so far. According to the previous report, allo-speci c anti-HLA antibody was di cult to detect in human immune system chimeric mouse model without humanized liver 21 . In our mouse model, we con rmed immune cell accumulation, especially in human hepatocyte areas and detection of transplanted human hepatocyte-speci c anti-HLA antibody.…”

Skin-Grafting and Dendritic Cell “Boosted” Humanized Mouse Models Allow the Pre-Clinical Evaluation of Therapeutic Cancer Vaccines

“…The expression of the α integrin CD11c on T cells has been linked to a highly activated T cell phenotype, which positively correlated in our animals with CD11b up-regulation, a sign of recent cell activation [ 24 ]. This model was also capable of sustaining high levels of B cells in the spleen, which have been previously reported to produce IgGs detectable in the blood of engrafted animals [ 42 ]. Notably, one of the B cell subsets present in the engrafted animals showed high expression of CD11c + .…”

Humanized mouse models for immuno-oncology research

“…Human CD19 + B cells are maintained at low levels in specific sites, including the spleen and bone marrow, for several weeks. In addition, human IgGs are detectable in the peripheral blood of Hu-PBL-SCID mice for their whole lifespan 55 .…”