Shared amino acid sequences in the NDβN and Nα regions of the T cell receptors of tumor‐infiltrating lymphocytes within malignant glioma

Abstract

The purpose of this study was to assess the V-(D)-J junctional region of the T cell receptor (TCR), the CDR3 region, which is responsible for glioma-specific antigen contact in alpha beta TCR-mediated recognition. We sequenced the TCR alpha and beta chains of V alpha 7, and V beta 13.1 cDNA derived from tumor-infiltrating lymphocytes (TIL) of 12 glioma patients and also the corresponding clones from the patients' peripheral blood lymphocytes (PBL). A shared V beta 13.1 DJ sequence of the CDR3 region, ND beta N, was demonstrated in 49 of 66 V beta 13.1+ clones (74.2%) from the glioma TIL, whereas only 4 of 33 clones (12.1%) were observed in the V beta 13.1+ clones from the PBL (p < 0.001). A common VDJ sequence, FCASS (V beta 13.1)-YRLPWGTSDS (ND beta N)-GELFF (J beta 2.2), was observed not only in the gliomas from each patient, but also among all the patients with a preference for V beta 13.1. In contrast, the amino acid sequences of the V beta 13.1+ PBL clones were diverse and random. Next, we sequenced subclones from other V beta subfamilies randomly selected to compare their VDJ region rearrangements (V beta 3 and V beta 5.1). In contrast to V beta 13.1, the amino acid sequences of these junctional regions were completely different in these subclones. The V-J junctional region of the alpha chain is dominated by a few clones in some patients, and no shared amino acid sequences were detected in the TCR V alpha junctional region. However, in the N alpha region of the V alpha 7-bearing TIL clones, arginine was used in 27 of 44 clones (61.4%) compared to only 3 of 12 clones from the PBL (p < 0.05). These results are consistent with the hypothesis that a clonal expansion/accumulation of glioma lineage-specific T cells occurred in vivo at the tumor site and that these T cells may be recognizing glioma-specific antigens.

Astrocytoma infiltrating lymphocytes include major T cell clonal expansions confined to the CD8 subset

“…many BV1 expansions with identical CDR3 size), but never conserved CDR3 sequences. This contrasts with previous TCR molecular analysis of astrocytoma infiltrating lymphocytes (but of non-Caucasian patients) reporting preferential AV7 and BV13 gene segment usage, with an identical BV13 transcript in different astrocytoma samples (53,54). In our series of Caucasian patients, this recurrent BV13 sequence was never detected in eight astrocytoma samples, including two patients expressing HLA-A24, the haplotype commonly (but not exclusively) associated with the recurrent TCR previously described (54) (G. Perrin, unpublished observations).…”

Antigen-loaded MR1 tetramers define T cell receptor heterogeneity in mucosal-associated invariant T cells

“…This noncanonical TRAJ usage was observed using either MR1-Ag tetramers or mAb TRAV1-2 + /CD161 hi markers to identify MAIT cells. TRAJ20 + MAIT cells have previously been reported (Gold et al, 2010, and although there have been other studies suggestive of repertoire diversity in MAIT cells (Ebato et al, 1994;Maru et al, 2003;Hwang et al, 2006), these cannot be confirmed in the absence of definitive MAIT cell characterization. Our study demonstrates that in some individuals, a surprisingly high frequency (up to 31%) of MR1-Ag tetramer + or mAb TRAV1-2 + cells can use alternative, non-TRAJ33 junctional genes.…”

TRAV gene expression in PBMCs and TILs in patients with breast cancer analyzed by a DNA melting curve (FQ-PCR) technique for TCR alpha chain CDR3 spectratyping

“…TCR CDR3 shows preferential usage TRBV or TRAV; There are common motifs in the CDR3 region or same CDR3 sequences [3,4]. A correlation between TCR beta chain CDR3 spectratyping and tumor characteristics have been reported for lung cancer [5,6], gastric cancer [7], colorectal cancer [8,9], melanoma [10], glioma [11], and uterine, and ovarian cancers [12]. There appears to be less correlation between TCR alpha-chain CDR3 spectratyping and tumor characteristics [3,13,14 ] .…”