title: TCR repertoire analysis by next generation sequencing allows complex differential diagnosis of T cell-related pathology.
abstract: Clonotype analysis is essential for complete characterization of antigen-specific T cells. Moreover, knowledge on clonal identity allows tracking of antigen-specific T cells in whole blood and tissue infiltrates and can provide information on antigenic specificity. Here, we developed a next generation sequencing (NGS)-based platform for the highly quantitative clonotype characterization of T cells and determined requirements for the unbiased characterization of the input material (DNA, RNA, ex vivo derived or cell culture expanded T cells). Thereafter we performed T cell receptor (TCR) repertoire analysis of various specimens in clinical settings including cytomegalovirus (CMV), polyomavirus BK (BKV) reactivation and acute cellular allograft rejection. Our results revealed dynamic nature of virus-specific T cell clonotypes; CMV reactivation was linked to appearance of new highly abundant antigen-specific clonalities. Moreover, analysis of clonotype overlap between BKV-, alloantigen-specific T cell-, kidney allograft- and urine-derived lymphocytes provided hints for the differential diagnosis of allograft dysfunction and enabled appropriate therapy adjustment. We believe that the established approach will provide insights into the regulation of virus-specific/anti-tumor immunity and has high diagnostic potential in the clinical routine.
authors.affiliation: Berlin-Brandenburg Center for Regenerative Therapies, Charité University Medicine Berlin, Berlin, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Dziubianau
authors.forename: J
authors.initials: J
authors.lastname: Hecht
authors.forename: L
authors.initials: L
authors.lastname: Kuchenbecker
authors.forename: A
authors.initials: A
authors.lastname: Sattler
authors.forename: U
authors.initials: U
authors.lastname: Stervbo
authors.forename: C
authors.initials: C
authors.lastname: Rödelsperger
authors.forename: P
authors.initials: P
authors.lastname: Nickel
authors.forename: A U
authors.initials: AU
authors.lastname: Neumann
authors.forename: P N
authors.initials: PN
authors.lastname: Robinson
authors.forename: S
authors.initials: S
authors.lastname: Mundlos
authors.forename: H-D
authors.initials: HD
authors.lastname: Volk
authors.forename: A
authors.initials: A
authors.lastname: Thiel
authors.forename: P
authors.initials: P
authors.lastname: Reinke
authors.forename: N
authors.initials: N
authors.lastname: Babel
mesh_terms: D001739:BK Virus / Q000235:genetics; D003587:Cytomegalovirus / Q000276:immunology* / Q000472:pathogenicity; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D003937:Diagnosis, Differential; D006084:Graft Rejection / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D016030:Kidney Transplantation / Q000009:adverse effects; D027601:Polyomavirus Infections / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D012189:Retrospective Studies; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology* / Q000821:virology; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D014775:Virus Activation
chemical_list: D011948:Receptors, Antigen, T-Cell
keywords: Acute cellular rejection; BKV; differential diagnosis; kidney transplantation; next generation sequencing; polyoma