Beryllium binding to HLA-DP molecule carrying the marker of susceptibilityto berylliosis glutamate beta 69

Berylliosis is a chronic granulomatous disorder caused by inhalation of Be dusts that is driven by the accumulation Be-specific CD-rif Th1-cells at di sease sites. Susceptibility to berylliosis has Lt en associated with the su pratypic variant of HLA-DP gene coding for glutamate at position beta 69 (H LA-DP beta Glu69). The aim of this study was to test the hypothesis that th e HLA-DP beta Glu69 residue plays a role in the interaction with Be. To thi s end, soluble HLA-DP2 molecule (carrying beta Glu69) and its mutated form carrying lysine at position beta 69 (HLA-DP2Lys69) were produced in Drosoph ila melanogaster and then used in a Be binding assays. BeSO4 (1-1000 muM) w as used to compete for the binding of the biotinilated invariant chain-deri ved peptide CLIP (50 muM). BeSO4 was capable of compete out biotin-CLIP bin ding from the HLA-DP2 (IC50%: 4.5 muM of BeSO4 at pH 5.0 and 5.5 muM of BeS O4 at pH 7.5), but not from the HLA-DP2Lys69 molecule (IC50%: 480 muM of Be SO4 at pH 5.0 and 220 muM of BeSO4 at pH 7.5). Moreover, the binding of NFL D.M60, a MoAb recognizing an epitope in the HLA-DP peptide binding region, to the HLA-DP2, but not to the HLA-DP2Lys69 soluble molecules was inhibited BeSO4. NFLD.M60 binding to HLA-DP2, bur not ro HLA-DP2Lys69 stably transfe cted murine cells was also inhibited Ly De both at pH 5.0 and at pH 7.5. Th e data indicate a direct interaction of Be with the HLA-DPGlu69 molecule, i n the absence of antigen processing. (C) American Society fur Histocompatib ility and Immunogenetics, 2001. Published by Elsevier Science Inc.