The goal of this grant has been to develop an approach to inducing mixed hematopoietic chimerism across MHC barriers with sufficiently high reliability and low toxicity that it could be used for the induction of organ allograft tolerance in humans. In the currently funded grant, we have utilized costimulatory blockade to reliably tolerize pre-existing peripheral and intrathymic CD4 and CDST cells to allogeneic marrow donors, achieving mixed chimerism and eventual central, deletional tolerance in the maintenance phase. Pre-existing peripheral and intrathymic mature alloreactive T cells are tolerized by allogeneic BMT with one injection of anti-GDI 54 mAb. Peripheral CDScells are more reliably tolerized if a low dose of TBI (3 Gy) is given 1 day prior to BMT. Peripheral CD4 cell tolerance involves anergy followed by deletion due to antigen presentation on APC lacking an activation signal through CD40. Donor-reactive peripheral CDS cells undergo even more rapid deletion than CD4 cells, and only CDStolerance, remarkably, requires recipient B cells and indirect allorecognition by recipient CD4 cells. Wehave obtained evidence fora novel indirect CDScell-mediated rejection pathway of cellular allograft rejection, which must be tolerized in our model. We now propose: 1)To identify molecular pathways through which BMTwith anti-CD154 leads to tolerization of peripheral donor- reactive CD4 Tcells. Theultimate goal of this aim is to identify reagents that could replace anti-CD154 by targeting pathways downstream of the blocked CD40/CD154 interaction;2)To identify the CD4/CD8/APC interactions leading to peripheral CD8cell tolerance when 3 Gy TBIon Day -1 and anti-CD154 are given. We hypothesize that separate interactions between directly alloreactive CD4and CDScells and indirectly alloreactive CD4and CDS cells and donor and host APC, respectively, tolerize two separate pathways of CDS-mediated marrow rejection;3)To identify the mechanisms by which recipient B cells promote tolerance of alloreactive CDS cells. We hypothesize that Ig receptors on recipient B cells recognizing donor alloantigens permit theconcentratedpresentation ofdonor MHC-derivedpeptides onboth class I and class II molecules of the B cell, which mediates tolerance of indirectly alloreactive CD4and CDS cells. Achievement of these goals will lead to a greater understanding of the pathways to allogeneic marrow rejection and tolerance and will allow the identification of molecular pathways downstream ofCD40 blockade to target in the development of new strategies for achieving this robust tolerance. These studies will ultimately enhance our ability to reliably achieve marrow engraftment without toxicity in patients needing organ transplants and those with hematologic disorders such as hemoglobinopathies.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL049915-17
Application #
7781341
Study Section
Transplantation, Tolerance, and Tumor Immunology (TTT)
Program Officer
Wagner, Elizabeth
Project Start
1994-06-01
Project End
2010-06-30
Budget Start
2010-03-01
Budget End
2010-06-30
Support Year
17
Fiscal Year
2010
Total Cost
$209,532
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Sykes, M (2015) Immune tolerance in recipients of combined haploidentical bone marrow and kidney transplantation. Bone Marrow Transplant 50 Suppl 2:S82-6
Haspot, F; Li, H W; Lucas, C L et al. (2014) Allospecific rejection of MHC class I-deficient bone marrow by CD8 T cells. Am J Transplant 14:49-58
Strober, Samuel; Spitzer, Thomas R; Lowsky, Robert et al. (2011) Translational studies in hematopoietic cell transplantation: treatment of hematologic malignancies as a stepping stone to tolerance induction. Semin Immunol 23:273-81
Lucas, Carrie L; Workman, Creg J; Beyaz, Semir et al. (2011) LAG-3, TGF-?, and cell-intrinsic PD-1 inhibitory pathways contribute to CD8 but not CD4 T-cell tolerance induced by allogeneic BMT with anti-CD40L. Blood 117:5532-40
Hermanrud, Christina E; Lucas, Carrie L; Sykes, Megan et al. (2011) Expression and purification of soluble murine CD40L monomers and polymers in yeast Pichia pastoris. Protein Expr Purif 76:115-20
Levesque, V; Bardwell, P D; Shimizu, I et al. (2011) B-cell-dependent memory T cells impede nonmyeloablative mixed chimerism induction in presensitized mice. Am J Transplant 11:2322-31
Mollov, J L; Lucas, C L; Haspot, F et al. (2010) Recipient dendritic cells, but not B cells, are required antigen-presenting cells for peripheral alloreactive CD8+ T-cell tolerance. Am J Transplant 10:518-526
Nikolic, Boris; Onoe, Takashi; Takeuchi, Yasuo et al. (2010) Distinct requirements for achievement of allotolerance versus reversal of autoimmunity via nonmyeloablative mixed chimerism induction in NOD mice. Transplantation 89:23-32
Fehr, Thomas; Lucas, Carrie L; Kurtz, Josef et al. (2010) A CD8 T cell-intrinsic role for the calcineurin-NFAT pathway for tolerance induction in vivo. Blood 115:1280-7
Sykes, Megan (2009) Mechanisms of transplantation tolerance in animals and humans. Transplantation 87:S67-9

Showing the most recent 10 out of 75 publications