title: Defining the alloreactive T cell repertoire using high-throughput sequencing of mixed lymphocyte reaction culture.
abstract: The cellular immune response is the most important mediator of allograft rejection and is a major barrier to transplant tolerance. Delineation of the depth and breadth of the alloreactive T cell repertoire and subsequent application of the technology to the clinic may improve patient outcomes. As a first step toward this, we have used MLR and high-throughput sequencing to characterize the alloreactive T cell repertoire in healthy adults at baseline and 3 months later. Our results demonstrate that thousands of T cell clones proliferate in MLR, and that the alloreactive repertoire is dominated by relatively high-abundance T cell clones. This clonal make up is consistently reproducible across replicates and across a span of three months. These results indicate that our technology is sensitive and that the alloreactive TCR repertoire is broad and stable over time. We anticipate that application of this approach to track donor-reactive clones may positively impact clinical management of transplant patients.
authors.affiliation: Adaptive Biotechnologies Corporation, Seattle, Washington, United States of America.
authors.forename: Ryan O
authors.initials: RO
authors.lastname: Emerson
authors.affiliation: Department of Surgery, Comprehensive Transplant center, Northwestern University, Chicago, Illinois, United States of America; Department of Microbiology-Immunology, Northwestern University, Chicago, Illinois, United States of America.
authors.forename: James M
authors.initials: JM
authors.lastname: Mathew
authors.affiliation: Department of Surgery, Comprehensive Transplant center, Northwestern University, Chicago, Illinois, United States of America.
authors.forename: Iwona M
authors.initials: IM
authors.lastname: Konieczna
authors.affiliation: Public Health Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.
authors.forename: Harlan S
authors.initials: HS
authors.lastname: Robins
authors.affiliation: Department of Surgery, Comprehensive Transplant center, Northwestern University, Chicago, Illinois, United States of America.
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Leventhal
mesh_terms: D000328:Adult; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D008297:Male; D016377:Organ Transplantation; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D014181:Transplantation Immunology / Q000235:genetics*
chemical_list: D011948:Receptors, Antigen, T-Cell