Restricted accessResearch articleFirst published online 2024-11
Knockdowns of CD3zeta Chain in Primary NK Cells Illustrate Modulation of Antibody-Dependent Cellular Cytotoxicity Against Human Immunodeficiency Virus-1
Multifaceted natural killer (NK) cell activities are indispensable for controlling human immunodeficiency virus (HIV)-1 transmission and pathogenesis. Among the diverse functions of NK cells, antibody-dependent cellular cytotoxicity (ADCC) has been shown to predict better HIV-1 protection. ADCC is initiated by the engagement of an Fc γ receptor CD16 with an Fc portion of the antibody, leading to phosphorylation of the CD3 ζ chain (CD3ζ) and Fc receptor γ chain (FcRγ) as well as downstream signaling activation. Though CD3ζ and FcRγ were thought to have overlapping roles in NK cell ADCC, several groups have reported that CD3ζ-mediated signals trigger a more robust ADCC. However, few studies have illustrated the direct contribution of CD3ζ in HIV-1-specific ADCC. To further understand the roles played by CD3ζ in HIV-1-specific ADCC, we developed a CD3ζ knockdown system in primary human NK cells. We observed that HIV-1-specific ADCC was inhibited by CD3ζ perturbation. In summary, we demonstrated that CD3ζ is important for eliciting HIV-1-specific ADCC, and this dynamic can be utilized for NK cell immunotherapeutics against HIV-1 infection and other diseases.
QiY, MartinMP, GaoX, et al.KIR/HLA pleiotropism: Protection against both HIV and opportunistic infections. PLoS Pathog, 2006; 2(8):e79; doi: 10.1371/journal.ppat.0020079
3.
VieillardV, Fausther-BovendoH, SamriA, et al.Specific phenotypic and functional features of natural killer cells from HIV-infected long-term nonprogressors and HIV controllers. J Acquir Immune Defic Syndr, 2010; 53(5):564–573; doi: 10.1097/QAI.0b013e3181d0c5b4
4.
AhmadF, HongHS, JackelM, et al.High frequencies of polyfunctional CD8+ NK cells in chronic HIV-1 infection are associated with slower disease progression. J Virol, 2014; 88(21):12397–12408; doi: 10.1128/JVI.01420-14
5.
WoolleyG, MosherM, KrollK, et al.Natural killer cells regulate acute SIV replication, dissemination, and inflammation, but do not impact independent transmission events. J Virol, 2022; 97(1):e0151922; doi: 10.1128/jvi.01519-22
6.
KrollKW, ShahSV, LucarOA, et al.Mucosal-homing natural killer cells are associated with aging in persons living with HIV. Cell Rep Med, 2022; 3(10):100773; doi: 10.1016/j.xcrm.2022.100773
7.
YatesNL, LiaoHX, FongY, et al.Vaccine-induced Env V1-V2 IgG3 correlates with lower HIV-1 infection risk and declines soon after vaccination. Sci Transl Med, 2014; 6(228):228ra39; doi: 10.1126/scitranslmed.3007730
8.
HessellAJ, HangartnerL, HunterM, et al.Fc receptor but not complement binding is important in antibody protection against HIV. Nature, 2007; 449(7158):101–104; doi: 10.1038/nature06106
9.
MichaelsenTE, AaseA, NorderhaugL, et al.Antibody dependent cell-mediated cytotoxicity induced by chimeric mouse-human IgG subclasses and IgG3 antibodies with altered hinge region. Mol Immunol, 1992; 29(3):319–326; doi: 10.1016/0161-5890(92)90018-s
10.
LanierLL, YuG, PhillipsJH. Analysis of Fc gamma RIII (CD16) membrane expression and association with CD3 zeta and Fc epsilon RI-gamma by site-directed mutation. J Immunol, 1991; 146(5):1571–1576.
11.
ShahSV, ManickamC, RamDR, et al.CMV primes functional alternative signaling in adaptive deltag NK cells but is subverted by lentivirus infection in rhesus macaques. Cell Rep, 2018; 25(10):2766–2774 e3; doi: 10.1016/j.celrep.2018.11.020
KwonHJ, KimHS. Signaling for synergistic activation of natural killer cells. Immune Netw, 2012; 12(6):240–246; doi: 10.4110/in.2012.12.6.240
14.
UpshawJL, SchoonRA, DickCJ, et al.The isoforms of phospholipase C-gamma are differentially used by distinct human NK activating receptors. J Immunol, 2005; 175(1):213–218; doi: 10.4049/jimmunol.175.1.213
15.
CellaM, FujikawaK, TassiI, et al.Differential requirements for Vav proteins in DAP10- and ITAM-mediated NK cell cytotoxicity. J Exp Med, 2004; 200(6):817–823; doi: 10.1084/jem.20031847
16.
ChenX, TrivediPP, GeB, et al.Many NK cell receptors activate ERK2 and JNK1 to trigger microtubule organizing center and granule polarization and cytotoxicity. Proc Natl Acad Sci U S A, 2007; 104(15):6329–6334; doi: 10.1073/pnas.0611655104
17.
ZhangW, Sloan-LancasterJ, KitchenJ, et al.LAT: The ZAP-70 tyrosine kinase substrate that links T cell receptor to cellular activation. Cell, 1998; 92(1):83–92; doi: 10.1016/s0092-8674(00)80901-0
18.
TingAT, KarnitzLM, SchoonRA, et al.Fc gamma receptor activation induces the tyrosine phosphorylation of both phospholipase C (PLC)-gamma 1 and PLC-gamma 2 in natural killer cells. J Exp Med, 1992; 176(6):1751–1755; doi: 10.1084/jem.176.6.1751
19.
BilladeauDD, BrumbaughKM, DickCJ, et al.The Vav-Rac1 pathway in cytotoxic lymphocytes regulates the generation of cell-mediated killing. J Exp Med, 1998; 188(3):549–559; doi: 10.1084/jem.188.3.549
20.
WatzlC, LongEO. Signal transduction during activation and inhibition of natural killer cells. Curr Protoc Immunol, 2010;Chapter 11:Unit 11 9B; doi: 10.1002/0471142735.im1109bs90
21.
LiuLL, LandskronJ, AskEH, et al.Critical Role of CD2 Co-stimulation in adaptive natural killer cell responses revealed in NKG2C-deficient humans. Cell Rep, 2016; 15(5):1088–1099; doi: 10.1016/j.celrep.2016.04.005
22.
Medjouel KhlifiH, GuiaS, VivierE, et al.Role of the ITAM-bearing receptors expressed by natural killer cells in cancer. Front Immunol, 2022; 13:898745; doi: 10.3389/fimmu.2022.898745
23.
LeeJ, ZhangT, HwangI, et al.Epigenetic modification and antibody-dependent expansion of memory-like NK cells in human cytomegalovirus-infected individuals. Immunity, 2015; 42(3):431–442; doi: 10.1016/j.immuni.2015.02.013
24.
PeppaD, Pedroza-PachecoI, PellegrinoP, et al.Adaptive reconfiguration of natural killer cells in HIV-1 infection. Front Immunol, 2018; 9:474; doi: 10.3389/fimmu.2018.00474
25.
HwangI, ZhangT, ScottJM, et al.Identification of human NK cells that are deficient for signaling adaptor FcRγ and specialized for antibody-dependent immune functions. Int Immunol, 2012; 24(12):793–802; doi: 10.1093/intimm/dxs080
26.
Suarez-FueyoA, BradleySJ, KatsuyamaT, et al.Downregulation of CD3zeta in NK cells from systemic lupus erythematosus patients confers a proinflammatory phenotype. J Immunol, 2018; 200(9):3077–3086; doi: 10.4049/jimmunol.1700588
27.
ZhangT, ScottJM, HwangI, et al.Cutting edge: Antibody-dependent memory-like NK cells distinguished by FcRγ deficiency. J Immunol, 2013; 190(4):1402–1406; doi: 10.4049/jimmunol.1203034
28.
LeeJ, ChangWLW, ScottJM, et al.FcRγ-NK cell induction by specific cytomegalovirus and expansion by subclinical viral infections in rhesus macaques. J Immunol, 2023; 211(3):443–452; doi: 10.4049/jimmunol.2200380
29.
ZhouJ, AmranFS, KramskiM, et al.An NK cell population lacking FcRγ is expanded in chronically infected HIV patients. J Immunol, 2015; 194(10):4688–4697; doi: 10.4049/jimmunol.1402448
30.
TuyishimeM, SprengRL, HueberB, et al.Multivariate analysis of FcR-mediated NK cell functions identifies unique clustering among humans and rhesus macaques. Front Immunol, 2023; 14:1260377; doi: 10.3389/fimmu.2023.1260377
31.
LiuW, ScottJM, LangguthE, et al.FcRγ gene editing reprograms conventional NK cells to display key features of adaptive human NK cells. iScience, 2020; 23(11):101709; doi: 10.1016/j.isci.2020.101709
32.
MizrahiO, Ish ShalomE, BaniyashM, et al.Quantitative flow cytometry: Concerns and recommendations in clinic and research. Cytometry B Clin Cytom, 2018; 94(2):211–218; doi: 10.1002/cyto.b.21515
33.
HuangRS, LaiMC, ShihHA, et al.A robust platform for expansion and genome editing of primary human natural killer cells. J Exp Med, 2021; 218(3); doi: 10.1084/jem.20201529
34.
DahlvangJD, DickJK, SangalaJA, et al.Ablation of SYK kinase from expanded primary human NK Cells via CRISPR/Cas9 enhances cytotoxicity and cytokine production. J Immunol, 2023; 210(8):1108–1122; doi: 10.4049/jimmunol.2200488
35.
Tremblay-McLeanA, CoenraadsS, KianiZ, et al.Expression of ligands for activating natural killer cell receptors on cell lines commonly used to assess natural killer cell function. BMC Immunol, 2019; 20(1):8; doi: 10.1186/s12865-018-0272-x
36.
StephensonKE, JulgB, TanCS, et al.Safety, pharmacokinetics and antiviral activity of PGT121, a broadly neutralizing monoclonal antibody against HIV-1: A randomized, placebo-controlled, phase 1 clinical trial. Nat Med, 2021; 27(10):1718–1724; doi: 10.1038/s41591-021-01509-0
37.
TuyishimeM, GarridoC, JhaS, et al.Improved killing of HIV-infected cells using three neutralizing and non-neutralizing antibodies. J Clin Invest, 2020; 130(10):5157–5170; doi: 10.1172/JCI135557
38.
LiuE, MarinD, BanerjeeP, et al.Use of CAR-transduced natural killer cells in CD19-positive lymphoid tumors. N Engl J Med, 2020; 382(6):545–553; doi: 10.1056/NEJMoa1910607
39.
FelicesM, LenvikTR, DavisZB, et al.Generation of BiKEs and TriKEs to improve NK cell-mediated targeting of tumor cells. Methods Mol Biol, 2016; 1441:333–346; doi: 10.1007/978-1-4939-3684-7_28
40.
ValleraDA, FelicesM, McElmurryR, et al.IL15 trispecific killer engagers (TriKE) make natural killer cells specific to CD33+ targets while also inducing persistence, In Vivo expansion, and enhanced function. Clin Cancer Res, 2016; 22(14):3440–3450; doi: 10.1158/1078-0432.CCR-15-2710
41.
BardhiA, WuY, ChenW, et al.Potent in vivo NK cell-mediated elimination of HIV-1-infected cells mobilized by a gp120-bispecific and hexavalent broadly neutralizing fusion protein. J Virol, 2017; 91(20); doi: 10.1128/JVI.00937-17
42.
PomeroyEJ, HunzekerJT, KluesnerMG, et al.A genetically engineered primary human natural killer cell platform for cancer immunotherapy. Mol Ther, 2020; 28(1):52–63; doi: 10.1016/j.ymthe.2019.10.009
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
