This 5 year competing application describes an opportunity to explore the long term outcomes of thymus transplantation in detail, with particular focus on the role of thymic activity and T cell function. Over the last 15 years, our laboratory has established that thymus transplantation is a successful therapeutic strategy for infants born with complete DiGeorge anomaly (cDGA) who are athymic and thus have a fatal immunodeficiency. Thymus transplantation results in host T cell reconstitution and survival of approximately 73% of subjects. We now will address unanswered questions about the basic biology of thymus transplantation in our long term survivors. Questions include the following, 1) why do T cell numbers remain in the 10th percentile after cDGA subjects develop circulating T cells and T cell function? How long does the thymus function after transplantation? 2) Do the peripheral T cells undergo premature senescence and die at higher rates than normal from apoptosis? 3) How are host T cells restricted to host HLA after development in an unmatched donor thymus? What recipient cells are present in the donor thymus after transplantation that can effect positive selection? To address these questions, we will pursue three aims. The first specific aim will assess the etiology of low T cell numbers after transplantation. Numbers of recent thymic emigrants (RTE) will be quantified by flow cytometry. Thymic output of RTE will be quantified by in vivo deuterated water loading in cDGA subjects. The intrathymic proliferation of thymocytes will be quantified by determination of the signal joint to ? T cell receptor rearrangement excision circle (TREC) ratios. These data will be evaluated to determine if thymic function decreases with time after transplantation. The RTE and TREC data from cDGA subjects will be compared to data from partial DGA and healthy children to assess relative thymic output in cDGA. The second specific aim will address the survival of peripheral T cells. Quantification of telomere length in naove CD4 cells will show if there is premature senescence of these cells. Studies of apoptosis will address whether peripheral apoptosis contributes to the low T cell numbers found in subjects with cDGA. The third specific aim will assess the binding of T cell receptors to HLA by tetramer experiments. The binding to class I and class II tetramers will be used to evaluate restriction to host and donor HLA. Affinity of binding to HLA Class I tetramers will be assessed for cDGA subjects with and without matching to the thymus donor and will be compared to the binding of subjects with partial DiGeorge anomaly. By comparing the outcomes of the experiments from cDGA subjects to those with partial DGA, who have lymphopenia, and to those of healthy age matched children, we can determine if thymus transplantation across an HLA barrier contributes to defects in T cell numbers after transplantation.

Public Health Relevance

Thymus transplantation is a promising therapy being developed for the fatal congenital anomaly, complete DiGeorge anomaly. Detailed evaluation of the research subjects will elucidate immune defects that remain in the subjects after transplantation which may allow improved medical management for these subjects and extension of this therapy to other patient groups. Project Narrative Thymus transplantation is a promising therapy being developed for the fatal congenital anomaly, complete DiGeorge anomaly. Detailed evaluation of the research subjects will elucidate immune defects that remain in the subjects after transplantation which may allow improved medical management for these subjects and extension of this therapy to other patient groups.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01AI047040-15
Application #
8711171
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Hayes, Deborah
Project Start
Project End
Budget Start
Budget End
Support Year
15
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Duke University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
City
Durham
State
NC
Country
United States
Zip Code
27705
Ciupe, Stanca M; Devlin, Blythe H; Markert, Mary Louise et al. (2013) Quantification of total T-cell receptor diversity by flow cytometry and spectratyping. BMC Immunol 14:35
Chinn, I K; Milner, J D; Scheinberg, P et al. (2013) Thymus transplantation restores the repertoires of forkhead box protein 3 (FoxP3)+ and FoxP3- T cells in complete DiGeorge anomaly. Clin Exp Immunol 173:140-9
Chinn, Ivan K; Markert, M Louise (2011) Induction of tolerance to parental parathyroid grafts using allogeneic thymus tissue in patients with DiGeorge anomaly. J Allergy Clin Immunol 127:1351-5
Markert, M Louise; Marques, Jose G; Neven, Benedicte et al. (2011) First use of thymus transplantation therapy for FOXN1 deficiency (nude/SCID): a report of 2 cases. Blood 117:688-96
Markert, M Louise; Devlin, Blythe H; McCarthy, Elizabeth A (2010) Thymus transplantation. Clin Immunol 135:236-46
Chinn, Ivan K; Olson, John A; Skinner, Michael A et al. (2010) Mechanisms of tolerance to parental parathyroid tissue when combined with human allogeneic thymus transplantation. J Allergy Clin Immunol 126:814-820.e8
Ciupe, Stanca M; Devlin, Blythe H; Markert, M Louise et al. (2009) The dynamics of T-cell receptor repertoire diversity following thymus transplantation for DiGeorge anomaly. PLoS Comput Biol 5:e1000396
Markert, M Louise; Devlin, Blythe H; Chinn, Ivan K et al. (2009) Thymus transplantation in complete DiGeorge anomaly. Immunol Res 44:61-70
Li, Bin; Li, Jie; Hsieh, Chia-San et al. (2009) Characterization of cultured thymus tissue used for transplantation with emphasis on promiscuous expression of thyroid tissue-specific genes. Immunol Res 44:71-83
Markert, M L; Devlin, B H; Chinn, I K et al. (2008) Factors affecting success of thymus transplantation for complete DiGeorge anomaly. Am J Transplant 8:1729-36

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