Since mixed chimerism has been shown in mouse and humanized mouse models (Project 3) to have the ability not only to induce central T cell tolerance, but also to prevent new antibody responses and turn off existing antibody responses, the establishment of xenogeneic mixed chimerism remains an attractive means for achieving xenograft tolerance. Before GalT-KO miniature swine were available, the failure of attempts to achieve mixed chimerism across the pig-to-primate barrier was attributed to high levels of primate natural antibodies to the Gal antigen. However, despite the absence of the Gal antigen on the cell surface of GalT-KO hematopoeitic cells (HSC), we have found that these cells are rapidly cleared from the circulation of baboons shortly after infusion, eluding the establishment of mixed chimerism. Our data indicate that two of the most important factors playing a role in this clearance are: 1) natural antibodies to non-Gal determinants, which are variably present in different baboons;and 2) absence of the species-specific cell-membrane protein, CD47, which is required to inhibit the innate immune system from rapidly clearing cells from the circulation. The goal of this proposal is to overcome these two barriers in order to establish mixed xenogeneic chimerism of GalT-KO HSC in baboons. Specifically, we will: 1) Optimize xenogeneic bone marrow engraftment by avoiding natural and induced anti-non-Gal antibodies;2) Test effect of human CD47 on engraftment of porcine marrow in baboons using GalT-KO pig bone marrow transduced with a human CD47 expression vector;and 3) Test the effect of GalT-KO pig HSC engraftment in baboons, either alone or in combination with vascularized porcine thymus transplantation, on the induction of tolerance of GalT-KO renal xenografts.
Aim 3 will apply the findings of Aims 1 and 2, as well as strategies developed in Project 1 of this Program Project, to the mixed chimerism approach for inducing transplantation tolerance to organ xenografts in this preclinical, discordant species combination. In addition, the experiments planned will provide basic information on xenogeneic stem cell engrafment and on the immunologic pathways responsible for xenogeneic rejection and tolerance induction in primates. As such, these studies should have both theoretical and practical implications for the eventual application of xenotransplantation as a clinical modality.

Public Health Relevance

Xenotransplantation remains the best near-term hope for alleviating the critical shortage of allogeneic organs today. However, the high level of immunosuppression required to avoid rejection of xenotransplants is likely to be too dangerous for clinical use. The goal of this project is to achieve tolerance of xenotransplants through mixed chimerism, and to thereby avoid the need for immunosuppressive drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI045897-12
Application #
8377257
Study Section
Special Emphasis Panel (ZAI1-QV-I)
Project Start
Project End
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
12
Fiscal Year
2012
Total Cost
$286,301
Indirect Cost
$84,581
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Sykes, Megan (2018) IXA Honorary Member Lecture, 2017: The long and winding road to tolerance. Xenotransplantation 25:e12419
Proto, Jonathan D; Doran, Amanda C; Subramanian, Manikandan et al. (2018) Hypercholesterolemia induces T cell expansion in humanized immune mice. J Clin Invest 128:2370-2375
Chen, Mo; Wang, Yuantao; Wang, Hui et al. (2018) Elimination of donor CD47 protects against vascularized allograft rejection in mice. Xenotransplantation :e12459
Watanabe, Hironosuke; Sahara, Hisashi; Nomura, Shunichiro et al. (2018) GalT-KO pig lungs are highly susceptible to acute vascular rejection in baboons, which may be mitigated by transgenic expression of hCD47 on porcine blood vessels. Xenotransplantation 25:e12391
Sachs, David H (2018) Transplantation tolerance through mixed chimerism: From allo to xeno. Xenotransplantation 25:e12420
Fishman, Jay A; Sachs, David H; Yamada, Kazuhiko et al. (2018) Absence of interaction between porcine endogenous retrovirus and porcine cytomegalovirus in pig-to-baboon renal xenotransplantation in vivo. Xenotransplantation 25:e12395
Mastroianni, Melissa; Ng, Zhi Yang; Goyal, Ritu et al. (2018) Topical Delivery of Immunosuppression to Prolong Xenogeneic and Allogeneic Split-Thickness Skin Graft Survival. J Burn Care Res 39:363-373
Tanabe, T; Watanabe, H; Shah, J A et al. (2017) Role of Intrinsic (Graft) Versus Extrinsic (Host) Factors in the Growth of Transplanted Organs Following Allogeneic and Xenogeneic Transplantation. Am J Transplant 17:1778-1790
Yamada, Kazuhiko; Sykes, Megan; Sachs, David H (2017) Tolerance in xenotransplantation. Curr Opin Organ Transplant 22:522-528
Holzer, Paul W; Leonard, David A; Shanmugarajah, Kumaran et al. (2017) A Comparative Examination of the Clinical Outcome and Histological Appearance of Cryopreserved and Fresh Split-Thickness Skin Grafts. J Burn Care Res 38:e55-e61

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