KIR3DL1 and HLA-B Density and Binding Calibrate NK Education and Response to HIV This information is current as Jeanette E. Boudreau, Tiernan J. Mulrooney, Jean-Benoît Le of September 25, 2021. Luduec, Edward Barker and Katharine C. Hsu J Immunol 2016; 196:3398-3410; Prepublished online 9 March 2016; doi: 10.4049/jimmunol.1502469 http://www.jimmunol.org/content/196/8/3398 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2016/03/08/jimmunol.150246 Material 9.DCSupplemental http://www.jimmunol.org/ References This article cites 72 articles, 37 of which you can access for free at: http://www.jimmunol.org/content/196/8/3398.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 25, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology KIR3DL1 and HLA-B Density and Binding Calibrate NK Education and Response to HIV Jeanette E. Boudreau,*,1 Tiernan J. Mulrooney,* Jean-Benoıˆt Le Luduec,* Edward Barker,† and Katharine C. Hsu*,‡,x NK cells recognize self-HLA via killer Ig-like receptors (KIR). Homeostatic HLA expression signals for inhibition via KIR, and downregulation of HLA, a common consequence of viral infection, allows NK activation. Like HLA, KIR are highly polymorphic, and allele combinations of the most diverse receptor–ligand pair, KIR3DL1 and HLA-B, correspond to hierarchical HIV control. We used primary cells from healthy human donors to demonstrate how subtype combinations of KIR3DL1 and HLA-B calibrate NK education and their consequent capacity to eliminate HIV-infected cells. High-density KIR3DL1 and Bw4-80I partnerships endow NK cells with the greatest reactivity against HLA-negative targets; NK cells exhibiting the remaining KIR3DL1/HLA-Bw4 combinations demonstrate intermediate responsiveness; and Bw4-negative KIR3DL1+ NK cells are poorly responsive. Cytotox- Downloaded from icity against HIV-infected autologous CD4+ T cells strikingly correlated with reactivity to HLA-negative targets. These findings suggest that the programming of NK effector function results from defined features of receptor and ligand subtypes. KIR3DL1 and HLA-B subtypes exhibit an array of binding strengths. Like KIR3DL1, subtypes of HLA-Bw4 are expressed at distinct, predictable membrane densities. Combinatorial permutations of common receptor and ligand subtypes reveal binding strength, receptor density, and ligand density to be functionally important. These findings have immediate implications for prognosis in patients with HIV infection. Furthermore, they demonstrate how features of KIR and HLA modified by allelic variation calibrate http://www.jimmunol.org/ NK cell reactive potential. The Journal of Immunology, 2016, 196: 3398–3410. atural killer cells weigh inhibitory input against acti- specific for self-HLA class I molecules readily respond to acti- vating signals to distinguish healthy from diseased cells vation signals on target cells when HLA is absent or downregu- N (1, 2). To simultaneously achieve self-tolerance and lated (3, 5–7). In contrast, uneducated NK cells lacking inhibitory sensitivity to unhealthy cells, NK cells rely on interaction between receptors for self–class I molecules require more potent stimula- cell surface killer Ig-like receptors (KIR) and their HLA class I tion, including inflammation or opsonizing Abs, to be activated for ligands. KIR receptors convey inhibitory signaling upon binding effector function, but their insensitivity to inhibition by class I by guest on September 25, 2021 to their HLA ligands, thereby preventing autoaggression. Of in- molecules is beneficial in conditions in which HLA expression terest, KIR and HLA interaction is also critical for NK education, persists (8–10). a poorly understood process that confers NK cell sensitivity to loss Among individuals with the same KIR and HLA genes, NK of normal HLA expression on diseased cells. Independent segre- populations frequently exhibit dramatic differences in inhibitory gation of KIR and HLA genes frequently leads to the same indi- sensitivities and reactive potentials, possibly attributable to sig- vidual harboring NK populations exhibiting KIR for self-HLA as nificant allelic variation for both receptor and ligand (11–14). well as NK populations exhibiting KIR for which the cognate li- KIR3DL1 and HLA-B form the most ancient and polymorphic gand is lacking (3, 4). Educated cells expressing KIR molecules receptor–ligand pair (15). Wide variation in KIR3DL1 and HLA-B alleles occurs throughout global populations, but both genes are maintained at nearly 100% frequency, underscoring a coevolution *Immunology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY 10065; †Department of Immunology and Microbiology, Rush that has selected for substantial subtype diversity (16). University Medical Center, Chicago, IL 60612; ‡Department of Medicine, Memorial x The 77 unique alleles of KIR3DL1 are classified into 4 subtypes Sloan Kettering Cancer Center, New York, NY 10065; and Weill Cornell Medical based on their surface expression density and sequence homology: College, New York, NY 10065 KIR3DS1, null (KIR3DL1-n), low (KIR3DL1-l), and high (KIR3DL1-h) 1Current address: Department of Microbiology and Immunology, Dalhousie Univer- sity, Halifax, NS, Canada. (17–19). Considerably more diverse, the HLA-B alleles can ORCIDs: 0000-0002-9187-7128 (J.E.B.); 0000-0002-0218-2771 (T.J.M.). nevertheless be clustered based on polymorphisms at positions Received for publication November 23, 2015. Accepted for publication February 8, 77–83 that define either the Bw6 epitope, which does not interact 2016. with any KIR, or the Bw4 epitope, a ligand for KIR3DL1 (20). This work was supported by National Institutes of Health Grants R01 HL088134, Based on a dimorphism (isoleucine versus threonine) at position U01 AI069197, P01 CA23766, P30 CA008748-46, P03 CA008748 (Memorial Sloan 80 that affects interaction with KIR3DL1, the HLA-Bw4 alleles Kettering Cancer Center Core Grant), and R01 AI065361 (to E.B.). can be segregated further into Bw4-80I or Bw4-80T subtypes. Address correspondence and reprint requests to Dr. Katharine C. Hsu, Department of Medicine, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, In experiments using transfectant systems and tetramer bind- NY 10065. E-mail address: [email protected] ing, specific combinations of KIR3DL1 and HLA-Bw4 subtypes The online version of this article contains supplemental material. exhibit different receptor–ligand binding affinities and inhibitory Abbreviations used in this article: KIR, killer Ig-like receptor; b2m, b2-microglobu- strengths (13, 14, 21). KIR3DS1 and KIR3DL1-n subtypes are lin; MFI, median fluorescence intensity; MSKCC, Memorial Sloan Kettering Cancer not known to engage Bw4 molecules on neighboring cells; Center; PCR-SSP, PCR with sequence-specific primers. however, specific peptides, including those from HIV, may fa- Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 cilitate engagement of KIR3DS1 by Bw4-80I (22). KIR3DL1-l www.jimmunol.org/cgi/doi/10.4049/jimmunol.1502469 The Journal of Immunology 3399 and -h subtypes, in contrast, bind both Bw4 subtypes, with -Bw6, -C1, and -C2 subtypes using the HLA Immunopolymorphism data- varying strengths. KIR3DL1*005, a common KIR3DL1-l iso- base version 3.14.0. KIR genotyping and KIR3DL1 subtyping were form, binds Bw4-80I and -80T tetramers with similar affinity performed as previously described (19, 31, 32). Individuals with KIR3DL1*002-group high alleles were examined using a Luminex-based (21). KIR3DL1-h, notably the common KIR3DL1*001 and *015 KIR SSO platform according to the manufacturer’s instructions. The probe isoforms, preferentially engage Bw4-80I over -80T tetramers recognition site for bead region 64 spans the codons 236–239 and specif- (13, 21, 23). The functional relevance of such preferential ically targets the alleles KIR3DL1*002 and KIR3DL1*054. KIR3DL1*054 binding remains to be determined in primary NK cells, in which is a rare allele, lacking entirely from at least two patient cohorts (33, 34). Therefore, individuals positive for bead region 64 by Luminex and additional factors, including receptor and ligand densities, might KIR3DL1*002-group typing by PCR with sequence-specific primers (PCR- influence cell–cell interactions and NK education. SSP) were assumed to exhibit KIR3DL1*002. KIR3DL1*002 and *015 Combinations of KIR3DL1 and HLA-B subtypes are associated represent a total of 70.2% of the alleles identified in the KIR3DL1*002 with distinct rates of disease progression in persons infected with HIV group by PCR-SSP. Therefore, alleles not identified as KIR3DL1*002 in (24). Notably, pairings of Bw4-80I with KIR3DL1-h, -n, or KIR3DS1 this group