Binding affinity independent contribution of peptide length to the stability of peptide-HLA-DR complexes in live antigen presenting cells.
The effect of peptide length on the stability of peptide-HLR-DR1 (DR1) complexes was analyzed using two peptide series of increasing length, each containing a 7mer core with five DR1-binding anchors, extended stepwise with Ala residues at the N- and C-terminus, respectively. The Ala extensions, although did not affect binding affinity, significantly increased the half lives of peptide-DR1 complexes (from 1.5 h up to 10 h) in live antigen presenting cells (APC). Flanking residues from position -2 to 0 and 8 to 11 were involved in the affinity-independent increase of complex stability. The shortest (8mer and 9mer) peptides, with in vivo half lives of <2.5 h, were unable to form stable complexes with DR1 in presence of HLA-DM (DM) molecules, and were poor competitors of antigen presentation. Longer peptides were resistant to DM-mediated unloading, and were efficient competitors of antigen presentation. Thus, DM appears to limit short peptides in establishing biologically relevant DR occupancy, despite their high binding affinity. In APC, stable complexes can form only with high affinity peptides of >9 residues, and the longevity of complexes seems to depend on full of occupation of the binding site.
Siklodi
B
B
Department of Immunology, Hoffmann-La Roche Inc., Nutley, New Jersey 07110-1199, USA.
Vogt
A B
AB
Kropshofer
H
H
Falcioni
F
F
Molina
M
M
Bolin
D R
DR
Campbell
R
R
Hämmerling
G J
GJ
Nagy
Z A
ZA
D000911:Antibodies, Monoclonal; D017951:Antigen Presentation; D000938:Antigen-Presenting Cells / Q000276:immunology*; D001665:Binding Sites; D002461:Cell Line, Transformed; D006681:HLA-D Antigens / Q000276:immunology; D015798:HLA-DR1 Antigen / Q000378:metabolism*; D006207:Half-Life; D000949:Histocompatibility Antigens Class II; D006801:Humans; D008213:Lymphocyte Activation; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry; D013601:T-Lymphocytes / Q000276:immunology
D000911:Antibodies, Monoclonal; C416189:H2-M antigens; D006681:HLA-D Antigens; C084245:HLA-DM antigens; D015798:HLA-DR1 Antigen; D000949:Histocompatibility Antigens Class II; D009842:Oligopeptides; D010446:Peptide Fragments