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Sections of
* Oncology,
Surgery, and
Pathology, Departments of Oncology and Surgical Sciences and
Pediatrics, University of Padova, Padova, Italy; and
¶ Division of Medical Oncology, University Hospital, Padova, Italy
It is widely accepted that the repertoire of Melan-A-specific T
cells naturally selected in melanoma patients is diverse and mostly
nonoverlapping among different individuals. To date, however, no
studies have addressed the TCR profile in different tumor sites and the
peripheral blood from the same patient. We compared the TCR usage of
Melan-A-specific T cells from different compartments of a single
melanoma patient to evaluate possible clonotype expansion or
preferential homing over a 4-mo follow-up period. Using HLA-A2 peptide
tetramers, CD8+ T cells recognizing the modified Melan-A
immunodominant ELAGIGILTV peptide were isolated from four metastatic
lesions resected from a single melanoma patient, and their TCR
repertoire was studied. A panel of T cell clones was generated by cell
cloning of tetramer-positive cells. Analysis of the TCR 
-chain V
segment and the complementarity-determining region 3 (CDR3) length and
sequence revealed a large diversity in the TCR repertoire, with only
some of the clones showing a partial conservation in the CDR3. A
similar degree of diversity was found by analyzing a number of T cell
clones obtained after sorting a Melan-A-specific population derived
from PBLs of the same patient after in vitro culture with the
immunodominant epitope. Moreover, clonotypes found at one site were not
present in another, suggesting the lack of expansion and circulation of
one or more clonotypes. Taken together, these results buttress the
notion that the CTLs recognizing the immunodominant Ag of Melan-A
comprise a high number of different clonotypic TCR, of which only some
exhibit common features in the CDR3.
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