Herpes simplex encephalitis is a life-threatening disease of the central nervous system caused by herpes simplex viruses (HSVs). Following standard of care with antiviral acyclovir treatment, most patients still experience various neurological sequelae. Here we characterize HSV-1 infection of human brain organoids by combining single-cell RNA sequencing, electrophysiology and immunostaining. We observed strong perturbations of tissue integrity, neuronal function and cellular transcriptomes. Under acyclovir treatment viral replication was stopped, but did not prevent HSV-1-driven defects such as damage of neuronal processes and neuroepithelium. Unbiased analysis of pathways deregulated upon infection revealed tumour necrosis factor activation as a potential causal factor. Combination of anti-inflammatory drugs such as necrostatin-1 or bardoxolone methyl with antiviral treatment prevented the damages caused by infection, indicating that tuning the inflammatory response in acute infection may improve current therapeutic strategies.

Viral lung infections are a never-ending threat to public health due to the emergence of new variants and their seasonal nature. While vaccines offer some protection, the need for effective antiviral drugs remains high. The existing research methods using 2D cell culture and animal models have their limitations. Human cell-based tissue engineering approaches hold great promise for bridging this gap. Here, we describe a microextrusion bioprinting approach to generate three-dimensional (3D) lung models composed of four cell types: endothelial cells, primary fibroblasts, macrophage cells, and epithelial cells. A549 and Calu-3 cells were selected as epithelial cells to simulate the cells of the lower and upper respiratory tract, respectively. Cells were bioprinted in a hydrogel consisting of alginate, gelatin, hyaluronic acid, collagen, and laminin-521. The models were cultured under air–liquid interface (ALI) conditions to further enhance their physiological relevance as lung cells. Their viability, metabolic activity, and expression of specific cell markers were analyzed during long-term culture for 21 days. The constructs were successfully infected with both a seasonal influenza A virus (IAV) and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) omicron variant, demonstrating their potential for studying diverse viral infections.

Immunocompromised patients, such as those undergoing hematopoietic stem cell or solid organ transplantation, are highly susceptible to viral complications. Given the limitations and side effects of available antiviral therapies, adoptive transfer of antiviral T cells offers a promising alternative by restoring immune defense. However, existing models for evaluating antiviral T cell therapies lack physiological relevance, limiting accurate predictions of efficacy and safety. There is a critical need for in vitro human infection platforms that support personalized assessment of therapeutic responses. To address this, we developed antiviral T cell products (TCPs) targeting Influenza A virus (IAV)-infected cells, alongside an autologous human induced pluripotent stem cell (iPSC)-derived 3D lung organoid infection platform. This model recapitulates key immunological responses and is compatible with a new 3D high-throughput, high-content imaging pipeline. Our study provides the first proof-of-concept for assessing T cell-mediated cytotoxicity in a 3D in vitro lung infection model, advancing personalized antiviral immunotherapy development.