title: Efficient identification of T-cell clones associated with graft-versus-host disease in target tissue allows for subsequent detection in peripheral blood.
abstract: Graft-versus-host disease (GVHD) causes severe morbidity and mortality in allogeneic haematopoietic stem cell transplantation (HSCT) because of destruction of recipient tissues by donor alloreactive T cells. We hypothesized that GVHD-specific T-cell clones are expanded within affected tissue of HSCT patients and can also be detected in blood at the time of active disease. A multiplex polymerase chain reaction (PCR) was used to amplify T-cell receptor (TCR) variable beta (VB) chain rearrangements in skin biopsies from eight allogeneic HSCT patients. Molecular analysis of the complementarity-determining region 3 (CDR3) of amplified products defined expanded, potentially disease-associated 'clonotypes' and enabled the design of clonotype-specific PCR assays. We detected immunodominant clones in seven of eight GVHD-positive skin biopsies. In serial skin biopsies from the same patient, the identical clone was found in each biopsy. In a patient who underwent two successive HSCTs from different donors, distinct clones were identified for each engraftment. Using clonotypic PCR assays, individual tissue-derived clones could be identified in peripheral blood samples obtained during active GVHD. We hypothesize that clonotypic sequences derived from target tissue can serve as markers for GVHD and may have utility in diagnosis and monitoring response to therapy, as well as enable future therapies targeted against pathogenic clones.
authors.affiliation: The Experimental Hematology and Hematopoiesis Section, Taussig Cancer Center, The Cleveland Clinic Foundation, Cleveland, OH 44195, USA.
authors.forename: Rose C
authors.initials: RC
authors.lastname: Beck
authors.forename: Marcin
authors.initials: M
authors.lastname: Wlodarski
authors.forename: Lukasz
authors.initials: L
authors.lastname: Gondek
authors.forename: Karl S
authors.initials: KS
authors.lastname: Theil
authors.forename: Ralph J
authors.initials: RJ
authors.lastname: Tuthill
authors.forename: Ronald
authors.initials: R
authors.lastname: Sobeck
authors.forename: Brian
authors.initials: B
authors.lastname: Bolwell
authors.forename: Jaroslaw P
authors.initials: JP
authors.lastname: Maciejewski
mesh_terms: D002999:Clone Cells / Q000276:immunology*; D022801:Complementarity Determining Regions / Q000235:genetics; D015329:Gene Rearrangement, T-Lymphocyte / Q000276:immunology; D006086:Graft vs Host Disease / Q000276:immunology*; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis / Q000235:genetics; D012867:Skin / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D022801:Complementarity Determining Regions; D016693:Receptors, Antigen, T-Cell, alpha-beta