TomotherapyandHematopoieticStemCellsforTolerancetoKidneyTransplants ABSTRACT The overarching goal of this project is to develop a tolerance induction protocol for MHC disparate kidney transplantsinrhesusmacaquesandtoelucidatetheunderlyingmechanismsoftheinductionandmaintenance ofmixedchimerismandtoleranceinthismodel.TheprimaryhypothesisisthattolerancetoMHCmismatched kidneytransplantscanbesafelyandeffectivelyachievedbyestablishingamixedchimericstateusinganewly established post?transplant non?myeloablative, helical tomotherapy?based total lymphoid irradiation (TLI)? based conditioning regimen followed by donor bone marrow?CD34+ hematopoietic cell (HSC) infusions. In addition, we will elucidate mechanisms of host immunoregulatory characteristics that are associated with successfulHSCengraftmentandmaintenanceofthechimericstate.Weproposetotestourhypothesesbymeans of 2 specific aims: 1.) Combined Bone Marrow Hematopoietic Cell/Kidney Transplants todetermine the proportionofrhesusmacaquerecipientsofdisparateunrelateddonorkidneytransplantsthatachievechimerism (without GVHD) and can be withdrawn from all immunosuppressive drugs for greater than 2 years while maintainingnormalallograftfunctionandwithoutrejection.Wewillmeasurethestateofmixedchimerismin recipientsasafunctionoftimepost?transplantandaccordingtovariousperipheralbloodandbonemarrowcell types using DNA (STR) analysis, and with flow cytometry using rhesus antibodies specific for MHC class I Mamuallelesofthedonor.2)ImmuneMonitoring,ImmunopathologyandImmunocompetence.Wewill:a.) determine early recipient immunoregulatory changes of host myeloid cells induced by the TLI?based conditioningregimenthatcorrelatewiththesuccessofbonemarrowengraftment,b.)determineifthemixed chimeric state induces changes of host dendritic cell acquisition of donor MHC class I antigen and PD?LI expression at serial time points during and after withdrawing immunosuppression, c.) characterize the developmentofrenalallograftimmuneandnon?immuneinjurybyanalysisofserialrenalallograftbiopsies,and d.)determinethedegreeofrecipientimmune?competencyafterimmunosuppressivedrugwithdrawalbytesting the recall T cell responses to cytomegalovirus antigens and tetanus toxoid. Knowledge gained through this rhesus tolerance induction protocol, including the underlying immunological mechanisms, will have direct relevance to a variety of deceased donor transplants. Furthermore, it will set a new course of healthcare innovationanddeliverythatwillgreatlybenefittransplantpatients,andotherpatientpopulations.

Public Health Relevance

/PUBLICHEALTHRELEVANCE Safelyandconsistentlyachievingoperationaltoleranceinhumantransplantrecipientswouldbeamagnificent breakthrough. This would render chronic immunosuppression (IS) exposure obsolete. There are great opportunities emerging that utilize several immune tolerance mechanisms to promote this challenging and highlybeneficialobjective.Oneofthemostpromisingiscreationofamixedchimericimmuneenvironmentin thehost,comprisingbothrecipientanddonorimmuneelements.Thiscanbeaccomplishedwithinfusionof donor hematopoietic stem cells (HSCs) and T cells to induce the recipient immune state tolerant to the transplantedsolidorganallowingISelimination.Theoverarchinggoalofthisprojectistodevelopasafeand effectiveimmunetoleranceinductionprotocolforMHCdisparatekidneytransplantationinrhesusmacaques andtoelucidatetheunderlyingmechanismsoftheinductionandmaintenanceoftoleranceinthismodel.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
2U01AI102456-06
Application #
9329967
Study Section
Special Emphasis Panel (ZAI1)
Program Officer
Shaw, Julia M
Project Start
2012-08-01
Project End
2022-07-31
Budget Start
2017-08-01
Budget End
2018-07-31
Support Year
6
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Surgery
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Haynes, Lynn D; Coonen, Jennifer; Post, Jennifer et al. (2017) Collection of hematopoietic CD34 stem cells in rhesus macaques using Spectra Optia. J Clin Apher 32:288-294
Sullivan, J A; Jankowska-Gan, E; Hegde, S et al. (2017) Th17 Responses to Collagen Type V, k?1-Tubulin, and Vimentin Are Present Early in Human Development and Persist Throughout Life. Am J Transplant 17:944-956
Zahorchak, A F; Ezzelarab, M B; Lu, L et al. (2016) In Vivo Mobilization and Functional Characterization of Nonhuman Primate Monocytic Myeloid-Derived Suppressor Cells. Am J Transplant 16:661-71
Fantus, Daniel; Rogers, Natasha M; Grahammer, Florian et al. (2016) Roles of mTOR complexes in the kidney: implications for renal disease and transplantation. Nat Rev Nephrol 12:587-609
Oliver, Stefan L; Yang, Edward; Arvin, Ann M (2016) Varicella-Zoster Virus Glycoproteins: Entry, Replication, and Pathogenesis. Curr Clin Microbiol Rep 3:204-215
Guo, H; Lu, L; Wang, R et al. (2016) Impact of Human Mutant TGF?1/Fc Protein on Memory and Regulatory T Cell Homeostasis Following Lymphodepletion in Nonhuman Primates. Am J Transplant 16:2994-3006
Wang, Zhaohui; Pratts, Shannon G; Zhang, Huiping et al. (2016) Treg depletion in non-human primates using a novel diphtheria toxin-based anti-human CCR4 immunotoxin. Mol Oncol 10:553-65
Kempton, Steve J; Robertson, Frederick A; Bentz, Michael L et al. (2015) Helical tomotherapy-based total lymphoid irradiation: a more precise and less toxic radiation delivery system for consideration in composite tissue allotransplantation. Plast Reconstr Surg 135:655e-656e
Ezzelarab, Mohamed B; Cooper, David K C; Thomson, Angus W (2015) Cell-based immunosuppression in kidney transplantation: the value of non-human primate studies. Kidney Int 88:1196-7
Burlingham, William J; Jankowska-Gan, Ewa; Kempton, Steve et al. (2015) Patterns of Immune Regulation in Rhesus Macaque and Human Families. Transplant Direct 1:e20