title: Single-cell analysis of the human NK cell response to missing self and its inhibition by HLA class I.
abstract: Natural killer (NK) cells activate quickly in response to pathogens, tumors, and allogeneic hematopoietic cell transplants. Modulating the NK cell response are clonally distributed NK cell receptors that survey cells for change in the expression of major histocompatibility complex (MHC) class I and structurally related ligands. Here the enzyme-linked immunospot (ELISPOT) assay, intracellular cytokine staining (ICS), and short-term culture were used to quantify the response of bulk NK cell populations from human donors to HLA class I-deficient 221 cells and to 221 cells transfected with single HLA class I allotypes. NK cells in cultures containing interleukin-2 (IL-2) or IL-12 exhibited specificities of HLA class I-mediated inhibition that correlated well with those previously defined using NK cell clones in long-term culture and with the frequencies of cells expressing particular inhibitory HLA class I receptors. Culture with IL-12, but not IL-2, gave an increased frequency of cells expressing CD94: NKG2A but no change in killer immunoglobulin-like receptor (KIR) expression. For some heterozygote combinations of KIR3DL1 alleles, ICS can be used to compare the functional properties of the 2 allotypes. Thus, both the low-expressing KIR3DL1*005 and the high-expressing KIR3DL1*002 gave similar inhibitory response on challenge with an HLA-B*5801 ligand. The single-cell assays developed here should facilitate future population study and clinical analysis of human NK cell regulation by MHC class I.
authors.affiliation: Department of Structural Biology, Sherman Fairchild Building, 299 Campus Dr West, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.
authors.forename: Monia
authors.initials: M
authors.lastname: Draghi
authors.forename: Nobuyo
authors.initials: N
authors.lastname: Yawata
authors.forename: Michael
authors.initials: M
authors.lastname: Gleimer
authors.forename: Makoto
authors.initials: M
authors.lastname: Yawata
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Valiante
authors.forename: Peter
authors.initials: P
authors.lastname: Parham
mesh_terms: D015703:Antigens, CD / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002478:Cells, Cultured; D003584:Cytological Techniques; D005838:Genotype; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D007157:Immunologic Surveillance; D018664:Interleukin-12 / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000276:immunology*; D037181:Lectins, C-Type / Q000032:analysis; D051920:NK Cell Lectin-Like Receptor Subfamily D; D011971:Receptors, Immunologic / Q000032:analysis / Q000235:genetics; D054340:Receptors, KIR; D054341:Receptors, KIR3DL1
chemical_list: D015703:Antigens, CD; D015395:Histocompatibility Antigens Class I; D007376:Interleukin-2; C515083:KIR3DL1 protein, human; C497869:KLRD1 protein, human; D037181:Lectins, C-Type; D051920:NK Cell Lectin-Like Receptor Subfamily D; D011971:Receptors, Immunologic; D054340:Receptors, KIR; D054341:Receptors, KIR3DL1; D018664:Interleukin-12