Visualization of inhibitory Ly49 receptor specificity with soluble major histocompatibility complex class I tetramers.
Murine natural killer (NK) cells are inhibited from killing their targets by the interaction between inhibitory, C-type lectin like Ly49 receptors and major histocompatibility complex (MHC) class I molecules. The receptors have overlapping specificity, and it has been difficult to analyze specific aspects of the interaction between different Ly49 receptors and their respective ligands. We have addressed this problem using tetramers of bacterially expressed, non-glycosylated, MHC class I molecules refolded with different peptides. Our results indicate that this technology is useful for analysis of Ly49 receptor specificity as well as for monitoring of NK cell subsets, with the following major conclusions emerging from this study: (1) tetramers of H-2D(d) bound the Ly49A receptor; the MHC associated glycan, previously suggested to be involved in recognition by this receptor, is thus not required for Ly49A receptor binding; (2) in support and extension of a recent report indicating peptide selectivity in the recognition of H-2K(b) by Ly49C(+) cells, H-2K(b) tetramer binding to Ly49C receptors was strongly influenced by the peptide presented by the MHC class I molecule; (3) tetramer binding allowed visualization of interactions that have not previously been detected in functional studies, such as the recognition of H-2D(b) by Ly49A and Ly49C.
Michaëlsson
J
J
Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden. Jakob.Michaelsson@mtc.ki.se
Achour
A
A
Salcedo
M
M
Kâse-Sjöström
A
A
Sundbäck
J
J
Harris
R A
RA
Kärre
K
K
D000818:Animals; D000950:Antigens, Ly; D002460:Cell Line; D006183:H-2 Antigens / Q000378:metabolism; D064532:Histocompatibility Antigen H-2D; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000378:metabolism*; D037181:Lectins, C-Type; D051379:Mice; D008807:Mice, Inbred BALB C; D055650:NK Cell Lectin-Like Receptor Subfamily A; D051381:Rats; D011971:Receptors, Immunologic / Q000378:metabolism*; D055630:Receptors, NK Cell Lectin-Like
D000950:Antigens, Ly; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D064532:Histocompatibility Antigen H-2D; D015395:Histocompatibility Antigens Class I; C527088:Klra1 protein, mouse; C527091:Klra3 protein, mouse; D037181:Lectins, C-Type; D055650:NK Cell Lectin-Like Receptor Subfamily A; D011971:Receptors, Immunologic; D055630:Receptors, NK Cell Lectin-Like