New parental cell lines for generating human hybridomas

Structural insight into MR1-mediated recognition of the mucosal associated invariant T cell receptor

“…It is thought that MAIT cells may be able to potentially recognize unknown microbial Ags presented by MR1 (Gold et al, 2010; Le Bourhis et al, 2010), although definitive identification of a bacterially encoded Ag is lacking at present. To explore the activation of human MAIT TCRs by MR1, we transduced genes for MAIT TCRs using TRBV20 (Vβ2), TRBV6-1 (Vβ13.3), or TRBV6-4 (Vβ13.5) β chains into either of the two human leukemia T cell lines SKW3 (Hundhausen et al, 1992; Gras et al, 2010) and Jurkat (Gillis and Watson, 1980), thus creating clonal cellular reagents for T cell recognition (SKW3.TRBV20, SKW3.TRBV6-1, and SKW3.TRBV6-4; Jurkat.BV20, Jurkat.BV6-1, and Jurkat.BV6-4). To generate Ag-presenting cells, HeLa cells and the MHC-I–deficient human lymphoblastoid cell line C1R were transduced with the gene for human MR1.…”

Allelic polymorphism in the T cell receptor and its impact on immune responses

“…1 d for the HLA-B*3501 HPVG multimer, no significant differences in CD69 up-regulation were observed between the different cell lines after peptide stimulation (not depicted). We reasoned that the high levels of CD8 expression on the JurkatCD8 cells could mask TCR–pMHC affinity differences, and therefore the SKW3 thymoma cell line, which is TCR α and β deficient and lacks high levels of CD8αβ expression (Hundhausen et al, 1992), was also transduced to express either the TK3 WT , TK3 Gln55His , or TK3 Gln55Ala TCRs. When these cell lines were tested in CD69 up-regulation experiments using various concentrations of the HPVGEADYFEY peptide presented on C1R–HLA-B*3501 APCs, the SKW3-TK3 Gln55His cell line was found to require much higher peptide concentrations for equivalent CD69 up-regulation in comparison to the SKW3-TK3 WT and SKW3-TK3 Gln55Ala cell lines (Fig.…”