Sorting and pooling strategy: a novel tool to map a virus proteome for CD8 T-cell epitopes.
Human cytomegalovirus (CMV) is a major cause of morbidity in immunocompromised individuals. However, no efficient vaccine has been developed to date. Identification of T-cell target proteins and epitopes is crucial not only for developing a successful immunization strategy, but also for new approaches using adoptive transfer of antigen-specific T-cells. The CMV genome has more than 200 open reading frames potentially coding for as many proteins. Here, we describe a robust, fast, and simple SPOT synthesis strategy, which allowed us to micro-synthesize every possible CD8 T-cell epitope in the entire potential CMV proteome. So far, 9069 of these peptides have been tested in an ex vivo T-cell stimulation assay. As well as confirming a number of previously known epitopes, we identified several new ones.
Ay
Bernhard
B
Institut für Medizinische Immunulogie, Charite - Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
Streitz
Mathias
M
Boisguerin
Prisca
P
Schlosser
Andreas
A
Mahrenholz
Carsten C
CC
Schuck
Sebastian D
SD
Kern
Florian
F
Volkmer
Rudolf
R
D000595:Amino Acid Sequence; D001704:Biopolymers / Q000737:chemistry; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003587:Cytomegalovirus / Q000235:genetics* / Q000276:immunology*; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000737:chemistry*; D005434:Flow Cytometry; D016679:Genome, Viral; D006801:Humans; D066298:In Vitro Techniques; D009842:Oligopeptides / Q000737:chemistry; D010449:Peptide Mapping; D020543:Proteome; D040901:Proteomics / Q000379:methods*
D001704:Biopolymers; D000939:Epitopes; D009842:Oligopeptides; D020543:Proteome