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The direct binding of a p58 killer cell inhibitory receptor to human histocompatibility leukocyte antigen (HLA)-Cw4 exhibits peptide selectivity.

Rajagopalan S, Long EO - J. Exp. Med. (1997)

Bottom Line: Binding of soluble KIR cl42 molecules to HLA-Cw4 expressed on transporter associated with antigen presentation (TAP)-deficient RMA-S cells occurred only upon exogenous peptide loading.Moreover, there was peptide selectivity in that certain substitutions at positions 7 and 8 of the nonamer peptide QYDDAVYKL abolished Cw4 interaction with KIR cl42 despite similar surface expression of HLA-C.The specificity of this direct interaction between peptide-loaded HLA-Cw4 on RMA-S cells and soluble KIR cl42 correlated with recognition by NK clones in that they were inhibited only by HLA-Cw4 loaded with the appropriate peptides.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland 20852, USA.

ABSTRACT
Natural killer (NK) cells in mice and humans express a number of structurally diverse receptors that inhibit target cell lysis upon recognition of major histocompatibility complex (MHC) class I molecules expressed on targets. The contribution of peptide to the structural features of class I required for NK cell inhibition appears to vary depending on the type of receptor engaged. Thus, while there is no peptide specificity in NK inhibition mediated by Ly-49A in the mouse, human histocompatibility antigen (HLA)-B*2705-specific NK clones displayed selectivity for peptides. In this report, we examine the role of peptide in the recognition of HLA-C by the defined killer cell inhibitory receptor (KIR) cl42 with established specificity for HLA-Cw4. Binding of soluble KIR cl42 molecules to HLA-Cw4 expressed on transporter associated with antigen presentation (TAP)-deficient RMA-S cells occurred only upon exogenous peptide loading. Moreover, there was peptide selectivity in that certain substitutions at positions 7 and 8 of the nonamer peptide QYDDAVYKL abolished Cw4 interaction with KIR cl42 despite similar surface expression of HLA-C. The specificity of this direct interaction between peptide-loaded HLA-Cw4 on RMA-S cells and soluble KIR cl42 correlated with recognition by NK clones in that they were inhibited only by HLA-Cw4 loaded with the appropriate peptides.

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Interaction of peptide-stabilized surface HLA-C with cl42– Ig. Cells were incubated with peptides (100 μM) for 24 h at 25°C and  tested for induction of HLA-C surface expression (closed bars) or KIR  cl42–Ig binding (open bars). The data are represented as mean fluorescence  intensity. The cl42–Ig binding to peptide Cw4#1 A5R was not done in  this particular experiment. In other experiments, binding was comparable  to that seen with peptide Cw4#1. Three other experiments gave similar  results.
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Figure 2: Interaction of peptide-stabilized surface HLA-C with cl42– Ig. Cells were incubated with peptides (100 μM) for 24 h at 25°C and tested for induction of HLA-C surface expression (closed bars) or KIR cl42–Ig binding (open bars). The data are represented as mean fluorescence intensity. The cl42–Ig binding to peptide Cw4#1 A5R was not done in this particular experiment. In other experiments, binding was comparable to that seen with peptide Cw4#1. Three other experiments gave similar results.

Mentions: TAP-deficient RMA-S cells previously transfected with human β2m (24) (referred to in this report as RMA-S) were used for transfections with HLA-Cw*0401 or HLA-Cw*0802 cDNA. HLA-Cw*0401 belongs to the group of HLA-C allotypes with amino acids N77K80, and HLA-Cw*0802 belongs to the group with amino acids S77N80. Transfectants were incubated at 25°C to faciliate surface expression of empty HLA-C molecules, screened by flow cytometry using the HLA-C-reactive mAb F4/326, and subcloned. One clone each, expressing high cell surface levels of either HLA-Cw4 or HLA-Cw8 at 25°C, was chosen for further study (Fig. 1). RMA-S-Cw4 transfectants were incubated with the peptide QYDDAVYKL, a high affinity consensus peptide that bound Cw*0401 (27). Peptide loading resulted in a dramatic increase in cell surface expression of HLA-Cw4 as depicted by the greater than fivefold increase in fluorescence intensity. This peptide did not affect the cell surface expression of HLA-Cw8 (see Fig. 2).


The direct binding of a p58 killer cell inhibitory receptor to human histocompatibility leukocyte antigen (HLA)-Cw4 exhibits peptide selectivity.

Rajagopalan S, Long EO - J. Exp. Med. (1997)

Interaction of peptide-stabilized surface HLA-C with cl42– Ig. Cells were incubated with peptides (100 μM) for 24 h at 25°C and  tested for induction of HLA-C surface expression (closed bars) or KIR  cl42–Ig binding (open bars). The data are represented as mean fluorescence  intensity. The cl42–Ig binding to peptide Cw4#1 A5R was not done in  this particular experiment. In other experiments, binding was comparable  to that seen with peptide Cw4#1. Three other experiments gave similar  results.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196286&req=5

Figure 2: Interaction of peptide-stabilized surface HLA-C with cl42– Ig. Cells were incubated with peptides (100 μM) for 24 h at 25°C and tested for induction of HLA-C surface expression (closed bars) or KIR cl42–Ig binding (open bars). The data are represented as mean fluorescence intensity. The cl42–Ig binding to peptide Cw4#1 A5R was not done in this particular experiment. In other experiments, binding was comparable to that seen with peptide Cw4#1. Three other experiments gave similar results.
Mentions: TAP-deficient RMA-S cells previously transfected with human β2m (24) (referred to in this report as RMA-S) were used for transfections with HLA-Cw*0401 or HLA-Cw*0802 cDNA. HLA-Cw*0401 belongs to the group of HLA-C allotypes with amino acids N77K80, and HLA-Cw*0802 belongs to the group with amino acids S77N80. Transfectants were incubated at 25°C to faciliate surface expression of empty HLA-C molecules, screened by flow cytometry using the HLA-C-reactive mAb F4/326, and subcloned. One clone each, expressing high cell surface levels of either HLA-Cw4 or HLA-Cw8 at 25°C, was chosen for further study (Fig. 1). RMA-S-Cw4 transfectants were incubated with the peptide QYDDAVYKL, a high affinity consensus peptide that bound Cw*0401 (27). Peptide loading resulted in a dramatic increase in cell surface expression of HLA-Cw4 as depicted by the greater than fivefold increase in fluorescence intensity. This peptide did not affect the cell surface expression of HLA-Cw8 (see Fig. 2).

Bottom Line: Binding of soluble KIR cl42 molecules to HLA-Cw4 expressed on transporter associated with antigen presentation (TAP)-deficient RMA-S cells occurred only upon exogenous peptide loading.Moreover, there was peptide selectivity in that certain substitutions at positions 7 and 8 of the nonamer peptide QYDDAVYKL abolished Cw4 interaction with KIR cl42 despite similar surface expression of HLA-C.The specificity of this direct interaction between peptide-loaded HLA-Cw4 on RMA-S cells and soluble KIR cl42 correlated with recognition by NK clones in that they were inhibited only by HLA-Cw4 loaded with the appropriate peptides.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland 20852, USA.

ABSTRACT
Natural killer (NK) cells in mice and humans express a number of structurally diverse receptors that inhibit target cell lysis upon recognition of major histocompatibility complex (MHC) class I molecules expressed on targets. The contribution of peptide to the structural features of class I required for NK cell inhibition appears to vary depending on the type of receptor engaged. Thus, while there is no peptide specificity in NK inhibition mediated by Ly-49A in the mouse, human histocompatibility antigen (HLA)-B*2705-specific NK clones displayed selectivity for peptides. In this report, we examine the role of peptide in the recognition of HLA-C by the defined killer cell inhibitory receptor (KIR) cl42 with established specificity for HLA-Cw4. Binding of soluble KIR cl42 molecules to HLA-Cw4 expressed on transporter associated with antigen presentation (TAP)-deficient RMA-S cells occurred only upon exogenous peptide loading. Moreover, there was peptide selectivity in that certain substitutions at positions 7 and 8 of the nonamer peptide QYDDAVYKL abolished Cw4 interaction with KIR cl42 despite similar surface expression of HLA-C. The specificity of this direct interaction between peptide-loaded HLA-Cw4 on RMA-S cells and soluble KIR cl42 correlated with recognition by NK clones in that they were inhibited only by HLA-Cw4 loaded with the appropriate peptides.

Show MeSH
Related in: MedlinePlus