A Genome-Wide CRISPR/Cas9-Based Screen Identifies Heparan Sulfate Proteoglycans as Ligands of Killer-Cell Immunoglobulin-Like Receptors

被引:3
|
作者
Klein, Klara [1 ,2 ,3 ]
Hoelzemer, Angelique [4 ,5 ,6 ]
Wang, Tim [1 ,2 ,7 ]
Kim, Tae-Eun [8 ]
Dugan, Haley L. [8 ,9 ]
Jost, Stephanie [8 ,10 ]
Altfeld, Marcus [4 ]
Garcia-Beltran, Wilfredo F. [8 ,11 ]
机构
[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA
[2] Whitehead Inst Biomed Res, 9 Cambridge Ctr, Cambridge, MA 02142 USA
[3] Univ Vet Med Vienna, Inst Pharmacol & Toxicol, Vienna, Austria
[4] Leibniz Inst Expt Virol, Hamburg, Germany
[5] Univ Med Ctr Eppendorf, Dept Internal Med 1, Div Infect Dis, Hamburg, Germany
[6] German Ctr Infect Res DZIF, Site Hamburg Lubeck Borstel Riems, Hamburg, Germany
[7] MIT, Dept Biol, Cambridge, MA USA
[8] Ragon Inst Massachusetts Gen Hosp MGH MIT & Harva, Cambridge, MA 02138 USA
[9] Adimab LLC, Lebanon, NH USA
[10] Beth Israel Deaconess Med Ctr, Ctr Virol & Vaccine Res, Boston, MA 02215 USA
[11] Massachusetts Gen Hosp MGH, Dept Pathol, Boston, MA 02114 USA
来源
FRONTIERS IN IMMUNOLOGY | 2021年 / 12卷
基金
美国国家卫生研究院;
关键词
CRISPR; screen; KIR; heparan sulfate; NK cells; HLA-F; BINDING; PROTEIN; MICE;
D O I
10.3389/fimmu.2021.798235
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
While human leukocyte antigen (HLA) and HLA-like proteins comprise an overwhelming majority of known ligands for NK-cell receptors, the interactions of NK-cell receptors with non-conventional ligands, particularly carbohydrate antigens, is less well described. We previously found through a bead-based HLA screen that KIR3DS1, a formerly orphan member of the killer-cell immunoglobulin-like receptor (KIR) family, binds to HLA-F. In this study, we assessed the ligand binding profile of KIR3DS1 to cell lines using Fc fusion constructs, and discovered that KIR3DS1-Fc exhibited binding to several human cell lines including ones devoid of HLA. To identify these non-HLA ligands, we developed a magnetic enrichment-based genome-wide CRISPR/Cas9 knock-out screen approach, and identified enzymes involved in the biosynthesis of heparan sulfate as crucial for the binding of KIR3DS1-Fc to K562 cells. This interaction between KIR3DS1 and heparan sulfate was confirmed via surface plasmon resonance, and removal of heparan sulfate proteoglycans from cell surfaces abolished KIR3DS1-Fc binding. Testing of additional KIR-Fc constructs demonstrated that KIR family members containing a D0 domain (KIR3DS1, KIR3DL1, KIR3DL2, KIR2DL4, and KIR2DL5) bound to heparan sulfate, while those without a D0 domain (KIR2DL1, KIR2DL2, KIR2DL3, and KIR2DS4) did not. Overall, this study demonstrates the use of a genome-wide CRISPR/Cas9 knock-out strategy to unbiasedly identify unconventional ligands of NK-cell receptors. Furthermore, we uncover a previously underrecognized binding of various activating and inhibitory KIRs to heparan sulfate proteoglycans that may play a role in NK-cell receptor signaling and target-cell recognition.
引用
收藏
页数:10
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