A NOVEL FIRST PRIMARY ANCHOR EXTENDS THE MHC-CLASS-II I-A(D)-BINDING-MOTIF TO ENCOMPASS 9 AMINO-ACIDS

The MHC class II molecule I-A(d) has been reported to bind peptides co ntaining a motif of six consecutive amino acids, We demonstrate that b inding of the murine IgG2a(b) heavy chain allopeptide gamma 2a(b) 435- 451 (Kabat numbering) to I-A(d) is strongly enhanced by a novel first primary anchor (P1) three residues N-terminal to this hexamer, This is based on flow cytometric assessment of the I-A(d) binding capacity of gamma 2a(b) peptide analogues, their antigenicity for I-A(d) restrict ed T cell clones and molecular modelling. The P1 pocket is broadly spe cific since aliphatic, aromatic, acidic, the basic histidine and small polar side chains all allowed good binding, By contrast, asparagine, arginine and glycine reduced the binding capacity 10-, 16- and >100-fo ld respectively, Truncation or glycine substitution at P1 decreased an tigenicity by a factor >1000, Nevertheless, I-A(d)-restricted T cells are not completely dependent on this anchor since high concentrations of a peptide with glycine-substituted P1 elicited maximal responses. A dditional anchoring side chains are found at P4, P6 and P9, The autolo gous IgG2a(a) heavy chain shares prominent epitopic residues with gamm a 2a(b) 435-451 at P3, P5 and P8. However, the lysine of gamma 2a(a) a t P9 impairs binding to I-A(d), which may explain why the gamma 2a(b) allopeptide-reactive T cells escaped negative selection, The data rati onalize our observation (Bartnes, K, and Hannestad, K. 1997. fur. J. I mmunol. 27:1124) that these T cells recognize a syngeneic B cell lymph oma, provided its presentation of intrinsic gamma 2a(a) is enhanced by surface IgG2a(a) ligation.