The role of the conserved residue in pocket A and the polymorphic residue in pocket E of HLA-B^*3501 in presentation of human minor histocompatibility peptides to T cells

Abstract

We investigated T cell recognition for human minor histocompatibility (hmH) peptides using HLA-B*3501 restricted, hmH specific cytotoxic T lymphocytes (CTL) clones. These CTL clones killed C1R cells expressing HLA-B*3501 but not C1R cells expressing chimeric antigens between HLA-B*3501 and HLA-B*5101. They also failed to kill C1R cells expressing HLA-B*3501 mutants at residue 152 (B*3501-V152E) or at residue 171 (B*3501-Y171H). The CTL clone failed to kill C1R cells expressing these mutant molecules loaded with the hmH peptides isolated from C1R-B*3501 cells although it killed a self-B cell line expressing HLA-B3501 loaded with the specific hmH peptides. The CTL clone also failed to kill T2 cells expressing the mutant molecules loaded with the specific peptides whereas it killed T2 cells expressing HLA-B*3501 loaded with the specific peptide. On the other hand, naturally occurring specific hmH peptides were isolated from purified B*3501-V152E and B*3501-Y171H molecules, indicating that both HLA-B*3501-V152E and HLA-B*3501-Y171H molecules can bind the hmH peptides. These findings indicate that both the conserved residue 171 in pocket A and the polymorphic residue 152 in pocket E are critical in recognition of the T cells but not binding of the hmH peptides. Furthermore, these results provide the possibility that the TCR recognizes a conformational structure of hmH peptides bound to HLA-B*3501 molecules.

Ex Vivo Profiling of CD8+-T-Cell Responses to Human Cytomegalovirus Reveals Broad and Multispecific Reactivities in Healthy Virus Carriers

“…T2 cells transfected with other individual HLA class I antigens were also used for MHC stabilization assays. T2 cells expressing HLA B35, B7, and B27 have been described elsewhere (44,46,53). T2 cells expressing HLA A3, A24, and B8 were established by transfecting expression vectors encoding individual class I alleles as described previously (24).…”

Peptides spanning the junctional region of both the abl/bcr and the bcr/abl fusion proteins bind common HLA class I molecules

“…27 T2 cells transfected with a number of different HLA alleles were kindly provided by Dr A McMichael, IMM, John Radcliffe Hospital, Oxford, UK (HLA-A*0301, -B*0702, -B*2705, -B*3501), and Dr M Masucci, MTC, Karolinska Institute, Stockholm, Sweden (HLA-A*1101). [28][29][30][31][32][33] The BM36.1 cell line is also defective in TAP function and has a similar phenotype as T2 with low expression of HLA class I (HLA-A*0101, HLA-B*3501) at the surface. 34 BM36.1 cells were kindly provided by Dr A Ziegler, Humboldt University, Berlin, Germany.…”

Autophagy mediates transporter associated with antigen processing-independent presentation of viral epitopes through MHC class I pathway

“…The T2.B35 cell line was derived by stable transfection of T2 cells with HLA B*3501. 13 Primary fibroblast cell lines were established from human foreskin fibroblasts. Immature monocyte-derived dendritic cells (MoDCs) were generated by plating peripheral blood mononuclear cells in 6-well plates for 90 minutes and treating the adherent monocytes with GM-CSF 1000 IU/mL and IL-4 1000 IU/mL (both from Schering-Plough) for 5 days.…”

“…14 In characterizing its endogenous presentation, we found that the LPL epitope was not only efficiently processed and presented by TAP-positive primary fibroblasts but also by TAP-deficient T2.B35 cells ( Figure 1A), the latter being stable HLA-B*3501 transfectants derived from a TAP1 and TAP2 deficient B xT hybrid cell line known as T2 or 174xCEM.T2. 13,16 We next used chemical inhibitors to determine whether antigen processing in the 2 cell types used the same or distinct pathways. The LPL epitope sequence is predicted to be hydrophobic.…”