Dass Debashree, Banerjee Anwesha, More Ashwini, Mukherjee Anupam
ICMR-National Institute of Translational Virology and AIDS Research, Pune, India.
Savitribai Phule Pune University, Pune, India.
Front Cell Infect Microbiol. 2025 May 29;15:1602965. doi: 10.3389/fcimb.2025.1602965. eCollection 2025.
Herpes Simplex Virus Type 2 is a prevalent sexually transmitted pathogen that causes genital herpes and severe neurological complications, including meningitis and encephalitis. A major challenge in HSV-2 infection is the uncontrolled inflammatory response mediated by NLRP3 inflammasome activation, leading to pyroptosis and excessive cytokine secretion. Despite its significant clinical burden, the molecular mechanisms underlying HSV-2-induced inflammation remain poorly understood. Recent evidence suggests that microRNAs play a crucial role in regulating host immune responses and inflammasome activation. In this study, we investigate the regulatory role of miR-141 and miR-211 in modulating inflammasome activation and viral replication during HSV-2 infection.
THP-1-derived macrophages were transfected with miR-141 or miR-211 mimics or scrambled controls before infection with HSV-2. Quantitative PCR and Western blot analysis were performed to assess the expression of NLRP3, CASP1, IL-1β, IL-18, and GSDM-D. Luciferase reporter assays were conducted to validate miRNA-target interactions, and ELISA was used to quantify cytokine levels in culture supernatants.
Our results demonstrate that HSV-2 infection significantly downregulates miR-141 and miR-211, leading to enhanced NLRP3 inflammasome activation, increased caspase-1 cleavage, and excessive secretion of IL-1β and IL-18, ultimately causing pyroptotic cell death. Transfection with miR-141 and miR-211 mimics restored miRNA expression, resulting in a marked suppression of inflammasome activation and inflammatory cytokine release, as well as significant inhibition of HSV-2 viral gene expression. Luciferase assays confirmed that miR-141 directly targets NLRP3, while miR-211 regulates CASP1, validating their roles as post-transcriptional repressors of inflammasome components.
These findings establish miR-141 and miR-211 as critical modulators of HSV-2-induced inflammasome activation, highlighting a novel miRNA-based regulatory mechanism. Restoring these miRNAs significantly reduces viral replication and inflammation, underscoring their potential as therapeutic targets for managing HSV-2-induced immunopathology. Future research should focus on validation and therapeutic optimization to develop miRNA-based interventions.
2型单纯疱疹病毒是一种常见的性传播病原体，可引起生殖器疱疹和严重的神经系统并发症，包括脑膜炎和脑炎。HSV-2感染中的一个主要挑战是由NLRP3炎性小体激活介导的不受控制的炎症反应，导致细胞焦亡和细胞因子过度分泌。尽管其临床负担很重，但HSV-2诱导炎症的分子机制仍知之甚少。最近的证据表明，微小RNA在调节宿主免疫反应和炎性小体激活中起关键作用。在本研究中，我们调查了miR-141和miR-211在HSV-2感染期间调节炎性小体激活和病毒复制中的作用。
在感染HSV-2之前，用miR-141或miR-211模拟物或乱序对照转染THP-1来源的巨噬细胞。进行定量PCR和蛋白质印迹分析以评估NLRP3、CASP1、IL-1β、IL-18和GSDM-D的表达。进行荧光素酶报告基因测定以验证miRNA-靶标相互作用，并使用ELISA定量培养上清液中的细胞因子水平。
我们的结果表明，HSV-2感染显著下调miR-141和miR-211，导致NLRP3炎性小体激活增强、caspase-1裂解增加以及IL-1β和IL-18过度分泌，最终导致细胞焦亡性细胞死亡。用miR-141和miR-211模拟物转染可恢复miRNA表达，导致炎性小体激活和炎性细胞因子释放明显受到抑制，以及HSV-2病毒基因表达受到显著抑制。荧光素酶测定证实miR-141直接靶向NLRP3，而miR-211调节CASP1，证实了它们作为炎性小体成分转录后抑制因子的作用。
这些发现确立了miR-141和miR-211是HSV-2诱导的炎性小体激活的关键调节因子，突出了一种基于miRNA的新型调节机制。恢复这些miRNA可显著减少病毒复制和炎症，强调了它们作为治疗HSV-2诱导的免疫病理学的治疗靶点的潜力。未来的研究应集中在验证和治疗优化上，以开发基于miRNA的干预措施。
