Mutational analysis of critical residues determining antigen presentation and activation of HLA-DQ0602 restricted T-cell clones.
Three different HLA-DQ0602 restricted T-lymphocyte clones (clones 5, 44, and 48) specific for two different Herpes simplex virus type 2 (HSV-2) VP16 peptides were used in a series of proliferation assays with BLS-1 cell lines expressing mutated HLA-DQ0604 molecules as APC. Up to four residues in the peptide-binding region of DQ0604 were replaced by the respective DQ0602 residue. For all three clones, residue beta70 played a crucial role in TCR recognition; beta30 and beta57 were important, although beta86 was less significant. Clone 5 and 48, specific to the HSV-2 VP16 369--379 peptide, responded to the same mutated DQ0604 molecules. Both clones could be stimulated only when the antigen presenting DQ molecule contained the DQ0602-like Gly at position beta70. Stimulation of clone 44, which recognized a different HSV-2 VP16 epitope (VP16 40-50), was less restricted. Molecular homology modeling showed that the beta70Arg of DQ0604 partially covered the peptide around P5/P6. Interactions of beta70 with residues from the antigen-peptide and polymorphic residues at positions beta30 and beta57 can modulate this effect. Supported by molecular modeling data, we conclude that the distinct molecular topography of DQ0602 is not contributed by a single residue, but rather the interactions of various polymorphic DQ residues with particular antigenic peptides.
Reichstetter
Sandra
S
Virginia Mason Research Center, Seattle, WA 98101, USA.
Papadopoulos
George K
GK
Moustakas
Antonis K
AK
Swanson
Eric
E
Liu
Andrew W
AW
Beheray
Sucheta
S
Ettinger
Ruth A
RA
Nepom
Gerald T
GT
Kwok
William W
WW
D017951:Antigen Presentation / Q000502:physiology; D002454:Cell Differentiation; D002478:Cells, Cultured; D002999:Clone Cells; D004252:DNA Mutational Analysis; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D006683:HLA-DQ Antigens / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D008213:Lymphocyte Activation; D008958:Models, Molecular; D011485:Protein Binding / Q000276:immunology; D017434:Protein Structure, Tertiary; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity; D017385:Sequence Homology; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
D018984:Epitopes, T-Lymphocyte; D006683:HLA-DQ Antigens; D011948:Receptors, Antigen, T-Cell