The Binding Site of NK Receptors on HLA-C Molecules Ofer Mandelboim, Hugh T Reyburn, Eric G Sheu, Mar Valés-Gómez, Daniel M Davis, Laszlo Pazmany, Jack L Strominger  Immunity  Volume 6, Issue 3, Pages 341-350 (March 1997) DOI: 10.1016/S1074-7613(00)80336-2

Figure 1 HLA-C Expression on the Various Transfectants 721.221 cells were transfected with wild-type HLA-Cw6 or HLA-Cw7 and with various HLA-Cw6 or HLA-Cw7 mutants at α76 (A), α73, and/or α90 (B). Transfections were performed as described in Experimental Procedures. Recipient cells (721.221) and the various transfectants were stained with W6/32 antibody, followed by fluorescein isothiocyanate (FITC)-labeled sheep anti–mouse immunoglobulin G (IgG) antibody (bold lines). Controls were the same cells stained with FITC anti–mouse IgG alone (light lines). Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)

Figure 2 NKIR Expression on NK Clones NK clones OM9, OM32, and OM49 were stained with the p58-specific MAbs EB6 (NKIR1, bold lines) and GL183 (NKIR2, dotted lines), followed by FITC anti–mouse IgG antibody (bold lines). Controls were the same cells stained with FITC anti–mouse IgG alone (light lines). Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)

Figure 3 αVal-76, the Locus-Specific Residue, of HLA-C Allotypes Is Essential for Inhibition of NK1- and NK2-Specific NK Cells (A) MV NK line (group 1 specificity); (B) OM32 clone (group 1 specificity); (C) OM NK line (group 2 specificity); (D) OM49 NK clone (group 2 specificity); (E) OM9 clone (group 1 and 2 specificity). NK lines and clones prepared from donor MV (A) or from donor OM (B–E) were incubated with various [35S]methionine-labeled target cells for 5 hr at various E:T ratios, with or without antibodies (GL183 or EB6 at 5 μg/ml). The targets are untransfected 721.221 cells or 721.221 cells transfected with the cDNAs shown. Only the data obtained at an E:T ratio of 10:1 (for the lines) and 5:1 (for the clones) are shown. One out of three representative experiments is shown. Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)

Figure 4 The Amino Acids at α73 and α90 Are Critical Components of the Interaction of HLA-C Allotypes with NK1- and NK2-Specific NK Cells (A) MV NK line (group 1 specificity); (B) OM32 clone (group 1 specificity); (C) OM NK line (group 2 specificity); (D) OM49 NK clone (group 2 specificity); (E) OM9 clone (group 1 and 2 specificity). For experimental details see the legend to Figure 3. Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)

Figure 5 Reversal of Inhibition Mutants Cw6/A73T,D90 (A) and Cw7/A73T,D90 (B) were used at different concentrations of the MAb EB6 and GL183. For experimental details see the legend to Figure 3. Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)

Figure 6 Cell Surface Expression of HLA-C Allotypes and Mutants at α78, α79, α81, and α82 For experimental details see the legend to Figure 1. Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)

Figure 7 α78, α79, α81, and α82 of HLA-C Allotypes Do Not Affect Recognition by NK1 and NK2 Clones For experimental details see the legend to Figure 3. Only an E:T ratio of 5:1 is shown. Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)

Figure 8 The Locations of the Four Key HLA-C Residues That Affect Its Interaction with NKIR1 and NKIR2 Residues α73, α76, α80, and α90 are highlighted on an α-carbon diagram of a class I MHC molecule, viewed from the top (Bjorkman et al. 1987). Immunity 1997 6, 341-350DOI: (10.1016/S1074-7613(00)80336-2)