Location
Moultrie, GA
Start Date
10-5-2021 12:00 AM
End Date
10-5-2021 12:00 AM
Description
Purpose of Study: Solid organ transplantation has been a life-saving procedure for thousands of patients worldwide. Recent advances on improving donor-screening diagnostics have aimed at identification of the most compatible donor for the transplant recipient to maximize allograft survival. Current standards of donor selection relies on HLA typing and in vitro mixed lymphocyte reaction (MLR) which do not take into account the in vivo environment and recipient’s adaptive immune response. Humanized mouse models are an appealing alternative that permits personalized investigation of the immunocompatibility of potential donor tissues for the recipient human immune system without putting patients at risk. By utilizing genomics, molecular and cellular analyses of allogeneic immune response we analyze the efficiency of our novel humanized mouse model to assess the donor-recipient compatibility and determine that it to be significantly more sensitive than conventional screening methods.
Methods Used: Human Leukocyte Antigen (HLA) typing and MLR for histocompatibility. Special strain of immunodeficient mice, NSG mice, subjected to irradiation (2Gy) and i.v injection of 8×10 peripheral blood mononuclear cells (PBMCs) from transplant recipients. For allogeneic immune response, humanized mice received 3×10 PBMCs from unrelated donors (UD) or related donors(RD). Whole genome transcriptome analysis and Real-Time PCR (RT-PCR) Transplant Rejection Array was used.
Summary of Results: Humanized mice demonstrated that allogeneic UD challenge induced significant splenomegaly with infiltration of activated cytotoxic human CD8+ CD25+ T cells expressing Perforin, Granzyme B and Interferon gamma (IFN-γ). Amongst the RDs, RD1 showed minimal allogeneic response while RD2 promoted higher cytotoxic CD8+ T cells infiltration, indicating that RD1 has better immunocompatibility with the recipient than RD2. However, MLR and HLA typing had failed to differentiate the 2 RDs showing them to have equal immunocompatibility with the recipient.
Conclusions: NSG-PBMC humanized mouse model was able to identify the related donor exhibiting minimal allogeneic response to the recipient. This model is significantly more immunologically sensitive than conventional MLR and HLA typing for selection of an immunocompatible donor for the transplant recipient.
Embargo Period
6-3-2021
Development of Humanized Mouse Model for Organ Transplantation
Moultrie, GA
Purpose of Study: Solid organ transplantation has been a life-saving procedure for thousands of patients worldwide. Recent advances on improving donor-screening diagnostics have aimed at identification of the most compatible donor for the transplant recipient to maximize allograft survival. Current standards of donor selection relies on HLA typing and in vitro mixed lymphocyte reaction (MLR) which do not take into account the in vivo environment and recipient’s adaptive immune response. Humanized mouse models are an appealing alternative that permits personalized investigation of the immunocompatibility of potential donor tissues for the recipient human immune system without putting patients at risk. By utilizing genomics, molecular and cellular analyses of allogeneic immune response we analyze the efficiency of our novel humanized mouse model to assess the donor-recipient compatibility and determine that it to be significantly more sensitive than conventional screening methods.
Methods Used: Human Leukocyte Antigen (HLA) typing and MLR for histocompatibility. Special strain of immunodeficient mice, NSG mice, subjected to irradiation (2Gy) and i.v injection of 8×10 peripheral blood mononuclear cells (PBMCs) from transplant recipients. For allogeneic immune response, humanized mice received 3×10 PBMCs from unrelated donors (UD) or related donors(RD). Whole genome transcriptome analysis and Real-Time PCR (RT-PCR) Transplant Rejection Array was used.
Summary of Results: Humanized mice demonstrated that allogeneic UD challenge induced significant splenomegaly with infiltration of activated cytotoxic human CD8+ CD25+ T cells expressing Perforin, Granzyme B and Interferon gamma (IFN-γ). Amongst the RDs, RD1 showed minimal allogeneic response while RD2 promoted higher cytotoxic CD8+ T cells infiltration, indicating that RD1 has better immunocompatibility with the recipient than RD2. However, MLR and HLA typing had failed to differentiate the 2 RDs showing them to have equal immunocompatibility with the recipient.
Conclusions: NSG-PBMC humanized mouse model was able to identify the related donor exhibiting minimal allogeneic response to the recipient. This model is significantly more immunologically sensitive than conventional MLR and HLA typing for selection of an immunocompatible donor for the transplant recipient.
Comments
Winner of 2021 Research Week Division of Research Award for Excellence in Research — SGA Campus