title: Clonotype analysis of cytomegalovirus-specific cytotoxic T lymphocytes.
abstract: Cytotoxic T lymphocytes (CTL) control the replication of human cytomegalovirus (CMV). Previous studies assessed the clonotypic composition of CTL specific for individual immunodominant peptides within a certain HLA context. Such an approach has inherent limitations and may not assess the true clonal CTL response in vivo. Here, the clonotypic composition of CMV-specific CTL was determined in HLA-A2, CMV-seropositive kidney transplant recipients and healthy blood donors after stimulation of peripheral blood mononuclear cells with either a pp65 whole-peptide pool or a single immunodominant peptide. Even after stimulation with the whole peptide pool, CMV-specific CTL remained monoclonal or oligoclonal. Regarding intraindividual variation, the CDR3 motifs of the dominant clones were identical to those observed in CTL generated by the single immunodominant peptide. Sequencing of the CDR3 regions demonstrated significant interindividual variation; however, structural homology was observed for immunodominant clonotypes in three individuals. In conclusion, the highly focused T cell receptor repertoire found after stimulation with either a single immunodominant peptide or a peptide pool demonstrates a pivotal role for immunodominant epitopes in the generation of a clonal repertoire. These results provide new insights into the regulation of CMV clonal dominance and may contribute to the design and monitoring of adoptive immunotherapy.
authors.affiliation: Experimental Haematology and Hematopoiesis Section, Taussig Cancer Center, Cleveland Clinic Foundation, Cleveland, Ohio, USA. nina.babel@charite.de
authors.forename: Nina
authors.initials: N
authors.lastname: Babel
authors.forename: Gordon
authors.initials: G
authors.lastname: Brestrich
authors.forename: Lukasz P
authors.initials: LP
authors.lastname: Gondek
authors.forename: Arne
authors.initials: A
authors.lastname: Sattler
authors.forename: Marcin W
authors.initials: MW
authors.lastname: Wlodarski
authors.forename: Nina
authors.initials: N
authors.lastname: Poliak
authors.forename: Nicole
authors.initials: N
authors.lastname: Bethke
authors.forename: Andreas
authors.initials: A
authors.lastname: Thiel
authors.forename: Markus H
authors.initials: MH
authors.lastname: Hammer
authors.forename: Petra
authors.initials: P
authors.lastname: Reinke
authors.forename: Jaroslaw P
authors.initials: JP
authors.lastname: Maciejewski
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D000941:Antigens / Q000737:chemistry; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D006680:HLA Antigens / Q000378:metabolism; D015789:HLA-A2 Antigen / Q000096:biosynthesis; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D016219:Immunotherapy, Adoptive / Q000379:methods; D016030:Kidney Transplantation / Q000276:immunology; D010455:Peptides / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism* / Q000821:virology*; D014763:Viral Matrix Proteins / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D006680:HLA Antigens; D015789:HLA-A2 Antigen; D010455:Peptides; D010750:Phosphoproteins; D011948:Receptors, Antigen, T-Cell; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa
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