Engagement of TLR3, TLR7, and NKG2D regulate IFN-γ secretion but not NKG2D-Mediated cytotoxicity by human NK cells stimulated with suboptimal doses of IL-12

被引:99
作者
Girart, Maria V.
Fuertes, Mercedes B.
Domaica, Carolina I.
Rossi, Lucas E.
Zwirner, Norberto W.
机构
[1] Hosp Clin Jose San Martin, Immunogenet Lab, Buenos Aires, DF, Argentina
[2] Univ Buenos Aires, Fac Med, Dept Microbiol, Buenos Aires, DF, Argentina
关键词
D O I
10.4049/jimmunol.179.6.3472
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
NK cells express different TLRs, such as TLR3, TLR7, and TLR9, but little is known about their role in NK cell stimulation. In this study, we used specific agonists (poly(I:C), loxoribine, and synthetic oligonucleotides containing unmethylated CpG sequences to stimulate human NK cells without or with suboptimal doses of IL-12, IL-15, or IFN-alpha, and investigated the secretion of IFN-gamma, cytotoxicity, and expression of the activating receptor NKG2D. Poly(I:C) and loxoribine, in conjunction with IL-12, but not IL-15, triggered secretion of IFN-gamma. Inhibition of IFN-gamma secretion by chloroquine suggested that internalization of the TLR agonists was necessary. Also, secretion of IFN-gamma was dependent on MEK1/ERK, p38 MAPK, p70(S6) kinase, and NF-kappa B, but not on calcineurin. IFN-alpha induced a similar effect, but promoted lesser IFN-gamma secretion. However, cytotoxicity (Cr-51 release assays) against MHC class I-chain related A (MICA)(-) and MICA(+) tumor targets remained unchanged, as well as the expression of the NKG2D receptor. Excitingly, IFN-gamma secretion was significantly increased when NK cells were stimulated with poly(I: C) or loxoribine and IL-12, and NKG2D engagement was induced by coculture with MICA(+) tumor cells in a PI3K-dependent manner. We conclude that resting NK cells secrete high levels of IFN-gamma in response to agonists of TLR3 or TLR7 and IL-12, and this effect can be further enhanced by costimulation through NKG2D. Hence, integration of the signaling cascades that involve TLR3, TLR7, IL-12, and NKG2D emerges as a critical step to promote IFN-gamma-dependent NK cell-mediated effector functions, which could be a strategy to promote Th1-biased immune responses in pathological situations such as cancer.
引用
收藏
页码:3472 / 3479
页数:8
相关论文
共 49 条
[1]   Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3 [J].
Alexopoulou, L ;
Holt, AC ;
Medzhitov, R ;
Flavell, RA .
NATURE, 2001, 413 (6857) :732-738
[2]   Interaction between conventional dendritic cells and natural killer cells is integral to the activation of effective antiviral immunity [J].
Andoniou, CE ;
van Dommelen, SLH ;
Voigt, V ;
Andrews, DM ;
Brizard, G ;
Asselin-Paturel, C ;
Delale, T ;
Stacey, KJ ;
Trinchieri, G ;
Degli-Esposti, MA .
NATURE IMMUNOLOGY, 2005, 6 (10) :1011-1019
[3]   Comparative analysis of human NK cell activation induced by NKG2D and natural cytotoxicity receptors [J].
André, P ;
Castriconi, R ;
Espéli, M ;
Anfossi, N ;
Juarez, T ;
Hue, S ;
Conway, H ;
Romagné, F ;
Dondero, A ;
Nanni, M ;
Caillat-Zucman, S ;
Raulet, DH ;
Bottino, C ;
Vivier, E ;
Moretta, A ;
Paul, P .
EUROPEAN JOURNAL OF IMMUNOLOGY, 2004, 34 (04) :961-971
[4]  
Bahram S, 2000, Adv Immunol, V76, P1
[5]   Activation of NK Cells and T Cells by NKG2D, a Receptor for Stress-Inducible MICA [J].
Bauer, Stefan ;
Groh, Veronika ;
Wu, Jun ;
Steinle, Alexander ;
Phillips, Joseph H. ;
Lanier, Lewis L. ;
Spies, Thomas .
JOURNAL OF IMMUNOLOGY, 2018, 200 (07) :2231-2233
[6]   Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition [J].
Bauer, S ;
Kirschning, CJ ;
Häcker, H ;
Redecke, V ;
Hausmann, S ;
Akira, S ;
Wagner, H ;
Lipford, GB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (16) :9237-9242
[7]   NKG2D-DAP10 triggers human NK cell-mediated killing via a Syk-independent regulatory pathway [J].
Billadeau, DD ;
Upshaw, JL ;
Schoon, RA ;
Dick, CJ ;
Leibson, PJ .
NATURE IMMUNOLOGY, 2003, 4 (06) :557-564
[8]   Synergy among receptors on resting NK cells for the activation of natural cytotoxicity and cytokine secretion [J].
Bryceson, YT ;
March, ME ;
Ljunggren, HG ;
Long, EO .
BLOOD, 2006, 107 (01) :159-166
[9]   Natural killer cell-mediated rejection of experimental human lung cancer by genetic overexpression of major histocompatibility complex class I chain-related gene A [J].
Busche, A ;
Goldmann, T ;
Naumann, U ;
Steinle, A ;
Brandau, S .
HUMAN GENE THERAPY, 2006, 17 (02) :135-146
[10]   Ectopic expression of retinoic acid early inducible-1 gene (RAE-1) permits natural killer cell-mediated rejection of a MHC class I-bearing tumor in vivo [J].
Cerwenka, A ;
Baron, JL ;
Lanier, LL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (20) :11521-11526