人类 Toll 样受体在自然发生的甲型流感感染中的作用。

Role of human Toll-like receptors in naturally occurring influenza A infections.

机构信息

Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Hong Kong, China.

出版信息

Influenza Other Respir Viruses. 2013 Sep;7(5):666-75. doi: 10.1111/irv.12109. Epub 2013 Apr 1.

DOI:10.1111/irv.12109

PMID:23552014

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5781199/

Abstract

BACKGROUND

We investigated the roles of Toll-like receptors (TLRs) in naturally occurring influenza.

METHODS

A prospective, case - control study was conducted. Adults hospitalized with virologically confirmed influenza A infections (onset <48 hours, before treatment) were compared with age-/gender-matched controls. TLRs (2, 3, 4, 7, 8, 9) expression in monocytes and dendritic cells (DCs - total, myeloid, plasmacytoid) was quantitated using flow cytometry. Gene expression of RLRs (RIG-1, MDA-5) was evaluated using real-time PCR. Concomitant signaling molecules expression, plasma cytokine/chemokine concentrations, and respiratory tract viral loads were measured. PBMCs were cultured and stimulated ex vivo with TLR-specific ligands for cytokine responses.

RESULTS

Forty two patients with influenza (24 A/H3N2, 18 A/H1N1pdm09) and 20 controls were studied. Patients' mean age was 68 ± 16 years; 81% had respiratory/cardiovascular complications. There were increased cellular expressions of TLR9, TLR8, TLR3, and TLR7 during influenza; TLR2 and TLR4 were suppressed. Results were similar for both virus strains. Higher TLR expression levels at presentation significantly correlated with lower viral loads (Spearman's rho: -0.46 to -0.69 for TLR9, TLR8, and TLR3; P-values <0.05). Multivariate regression models (adjusted for age, comorbidity, disease severity, time from onset) confirmed their independent associations. Increased signaling molecules (phospho-MAPKs, IκB) and inflammatory cytokines (IL-6, sTNFR-1, CCL2/MCP-1; CXCL10/IP-10, IFN-γ) correlated with increased TLR expression. RLRs were upregulated simultaneously. PBMCs of patients with influenza showed significant, dynamic changes in their cytokine responses upon TLR stimulation, compared with controls.

CONCLUSIONS

Our results suggest that TLRs play an important role in early, innate viral inhibition in naturally occurring influenza. Inflammatory cytokine responses are concomitantly induced. These findings support investigation of TLR targeting as a novel intervention approach for prophylaxis against influenza.

摘要

背景

我们研究了 Toll 样受体（TLR）在自然发生的流感中的作用。

方法

进行了一项前瞻性病例对照研究。将因病毒学证实的甲型流感感染（发病 <48 小时，治疗前）住院的成年人与年龄/性别匹配的对照进行比较。使用流式细胞术定量检测单核细胞和树突状细胞（DC）中 TLR（2、3、4、7、8、9）的表达。使用实时 PCR 评估 RLR（RIG-1、MDA-5）的基因表达。同时测量信号分子表达、血浆细胞因子/趋化因子浓度和呼吸道病毒载量。体外培养 PBMC 并使用 TLR 特异性配体刺激以进行细胞因子反应。

结果

共研究了 42 例流感患者（24 例 A/H3N2、18 例 A/H1N1pdm09）和 20 名对照。患者的平均年龄为 68 ± 16 岁；81%有呼吸道/心血管并发症。流感期间细胞表达 TLR9、TLR8、TLR3 和 TLR7 增加；TLR2 和 TLR4 受到抑制。两种病毒株的结果均相似。发病时更高的 TLR 表达水平与更低的病毒载量显著相关（Spearman's rho：TLR9、TLR8 和 TLR3 为-0.46 至-0.69；P 值<0.05）。多变量回归模型（调整年龄、合并症、疾病严重程度、发病时间）证实了它们的独立关联。增加的信号分子（磷酸化 MAPK、IκB）和炎症细胞因子（IL-6、sTNFR-1、CCL2/MCP-1；CXCL10/IP-10、IFN-γ）与 TLR 表达增加相关。RLR 同时上调。与对照组相比，流感患者的 PBMC 在 TLR 刺激下表现出明显的、动态的细胞因子反应变化。