The leading cause of mortality from COVID-19 infection is respiratory distress due to an exaggerated host immune response, resulting in hyper-inflammation and ensuing cytokine storms in the lungs. Current drug-based therapies are of limited efficacy, costly, and have potential negative side effects. By contrast, photobiomodulation therapy, which involves periodic brief exposure to red or infrared light, is a noninvasive, safe, and affordable method that is currently being used to treat a wide range of diseases with underlying inflammatory conditions. Here, we show that exposure to two 10-min, high-intensity periods per day of infrared light causes a marked reduction in the TLR-4 dependent inflammatory response pathway, which has been implicated in the onset of cytokine storms in COVID-19 patients. Infrared light exposure resulted in a significant decline in NFkB and AP1 activity as measured by the reporter gene assay; decreased expression of inflammatory marker genes IL-6, IL-8, TNF-alpha, INF-alpha, and INF-beta as determined by qPCR gene expression assay; and an 80% decline in secreted cytokine IL6 as measured by ELISA assay in cultured human cells. All of these changes occurred after only 48 hours of treatment. We suggest that an underlying cellular mechanism involving modulation of ROS may downregulate the host immune response after Infrared Light exposure, leading to decrease in inflammation. We further discuss technical considerations involving light sources and exposure conditions to put these observations into potential clinical use to treat COVID-19 induced mortality.

The leading cause of mortality from SARS-CoV-2 is an exaggerated host immune response, triggering cytokine storms, multiple organ failure and death. Current drug- and vaccine-based therapies are of limited efficacy against novel viral variants. Infrared therapy is a non-invasive and safe method that has proven effective against inflammatory conditions for over 100 years. However, its mechanism of action is poorly understood and has not received widespread acceptance. We herein investigate whether near-infrared (NIR) light exposure in human primary alveolar and macrophage cells could downregulate inflammatory cytokines triggered by the SARS-CoV-2 spike (S) protein or lipopolysaccharide (LPS), and via what underlying mechanism. Our results showed a dramatic reduction in pro-inflammatory cytokines within days of NIR light treatment, while anti-inflammatory cytokines were upregulated. Mechanistically, NIR light stimulated mitochondrial metabolism, induced transient bursts in reactive oxygen species (ROS) and activated antioxidant gene transcription. These, in turn, downregulated ROS and inflammatory cytokines. A causal relationship was shown between the induction of cellular ROS by NIR light exposure and the downregulation of inflammatory cytokines triggered by SARS-CoV-2 S. If confirmed by clinical trials, this method would provide an immediate defense against novel SARS-CoV-2 variants and other inflammatory infectious diseases.

Radiofrequency fields in the 1–28 GHz range are ubiquitous in the modern world, giving rise to numerous studies of potential health risks such as cancer, neurological conditions, reproductive risks and electromagnetic hypersensitivity. However, results are inconsistent due to a lack of precision in exposure conditions and vastly differing experimental models, whereas measured RF effects are often indirect and occur over many hours or even days. Here, we present a simplified RF exposure protocol providing a single 1.8 GHz carrier frequency to human HEK293 cell monolayer cultures. A custom-built exposure box and antenna maintained in a fully shielded anechoic chamber emits discrete RF signals which can be precisely characterized and modelled. The chosen amplitudes are non-thermal and fall within the range of modern telecommunication devices. A critical feature of the protocol is that cell cultures are exposed to only a single, short (15 min) RF exposure period, followed by detection of immediate, rapid changes in gene expression. In this way, we show that modulation of genes implicated in oxidative stress and ROS signaling is among the earliest cellular responses to RF exposure. Moreover, these genes respond in complex ways to varying RF signal amplitudes consistent with a hormetic, receptor-driven biological mechanism. We conclude that induction of mild cellular stress and reactive oxygen species (ROS) is a primary response of human cells to RF signals, and that these responses occur at RF signal amplitudes within the range of normal telecommunications devices. We suggest that this method may help provide a guideline for greater reliability and reproducibility of research results between labs, and thereby help resolve existing controversy on underlying mechanisms and outcomes of RF exposure in the general population.

Epstein–Barr virus nuclear antigen 1 (EBNA-1) is essential for viral episome maintenance and is consistently expressed in all forms of EBV latency. In a previous study, we observed EBNA-1 localizing in the nucleolus, as well as its interaction with the nucleolar protein EBP2 in both the nucleoplasm and nucleoli. Nucleoli are dynamic nuclear domains that coordinate ribosome assembly with the regulation of cell cycle progression and stress responses. Here, we studied the mechanism by which EBNA-1 achieves nucleolar localization and functional consequences for host cell physiology. We observed that EBNA-1 nucleolar accumulation occurs specifically during late G1 and S phase and depends on a bipartite nucleolar localization signal (NoLS) composed of two cooperative Weber motifs separated by 24 amino acids. Mutations disrupting NoLS abolish interaction with the nucleolar protein EBP2, whose depletion similarly prevents EBNA-1 nucleolar import. Once in the nucleolus, EBNA1 triggers a 50 % reduction in the synthesis of ribosomal RNA (rRNA) which leads to a global reduction in cellular protein production. In addition, EBNA1 nucleolar targeting induces reactive oxygen species (ROS) production and antioxidant treatment restores both rRNA transcription and protein synthesis. These findings identify EBP2 as a viral nucleolar docking partner and establish nucleolar targeting of EBNA1 as a regulated, cell cycle–dependent process that links oxidative stress to the perturbation of ribosome biogenesis. This mechanism may contribute to EBNA1’s ability to promote cell survival and transformation. IMPORTANCE: EBNA1 is the only Epstein–Barr virus protein that is consistently expressed in all forms of latency and in every EBV-associated tumor. Although it localizes to the nucleolus and interacts with three nucleolar proteins, its influence on nucleolar functions had never been investigated. Here, we demonstrate that EBNA1’s nucleolar targeting is a strictly cell-cycle–regulated process that depends on a previously unrecognized bipartite nucleolar localization signal as well as its interaction with the nucleolar protein EBP2. Once inside the nucleolus, EBNA1 induces a ROS-dependent suppression of rRNA synthesis, resulting in a global decrease in cellular protein production. In addition, these findings uncover a new function of EBNA1: the ability to induce nucleolar stress without activating canonical apoptotic pathways, thereby promoting host-cell survival and potentially contributing to EBV-driven oncogenesis