Alpha-lipoic acid (ALA) is a natural antioxidant dithiol compound, exerting antiproliferative and antimetastatic effects in various cancer cell lines. In our study, we demonstrated that ALA reduces the cell growth of prostate cancer cells LNCaP and DU-145. Western blot results revealed that in both cancer cells, ALA, by upregulating pmTOR expression, reduced the protein content of two autophagy initiation markers, Beclin-1 and MAPLC3. Concomitantly, MTT assays showed that chloroquine (CQ) exposure, a well-known autophagy inhibitor, reduced cells’ viability. This was more evident for treatment using the combination ALA + CQ, suggesting that ALA can reduce cells’ viability by inhibiting autophagy. In addition, in DU-145 cells we observed that ALA affected the oxidative/redox balance system by deregulating the KEAP1/Nrf2/p62 signaling pathway. ALA decreased ROS production, SOD1 and GSTP1 protein expression, and significantly reduced the cytosolic and nuclear content of the transcription factor Nrf2, concomitantly downregulating p62, suggesting that ALA disrupted p62-Nrf2 feedback loop. Conversely, in LNCaP cells, ALA exposure upregulated both SOD1 and p62 protein expression, but did not affect the KEAP1/Nrf2/p62 signaling pathway. In addition, wound-healing, Western blot, and immunofluorescence assays evidenced that ALA significantly reduced the motility of LNCaP and DU-145 cells and downregulated the protein expression of TGFβ1 and vimentin and the deposition of fibronectin. Finally, a soft agar assay revealed that ALA decreased the colony formation of both the prostate cancer cells by affecting the anchorage independent growth. Collectively, our in vitro evidence demonstrated that in prostate cancer cells, ALA reduces cell growth and counteracts both migration and invasion. Further studies are needed in order to achieve a better understanding of the underlined molecular mechanisms.

Background: Vitamin D is a pleiotropic steroid hormone with immunomodulatory properties. The role of the active metabolite, 1α,25-dihydroxyvitamin D ₃ (1,25(OH) ₂ vitamin D), in regulating human T-cell effector function and immune checkpoint (IC) pathways remains poorly defined. Methods: Peripheral blood mononuclear cells (PBMCs) from healthy donors were cultured for five days in presence or absence of 1,25(OH) ₂ vitamin D (10 or 100 nM). IFN-γ production was assessed by ELISpot assay. Baseline plasma 1,25(OH) ₂ vitamin D levels were measured. IC-related gene expression was evaluated by RT-qPCR. Flow cytometry was used to assess the expression of IC, degranulation, and activation markers. FluoroSpot assays were performed to quantify interleukin secretion. Results: 1,25(OH) ₂ vitamin D reduced IFN-γ expression and secretion independently of baseline circulating 1,25(OH) ₂ vitamin D levels. Dose-specific modulation of IC genes was detected, with increased PDCD1 and CTLA4 mRNA expression. Flow cytometry showed selective modulation within the CD8 ⁺ T-cell compartment, with increased PD-1 expression at 10 nM and increased CTLA-4 expression at 100 nM, together with decreased degranulation and activation markers. No significant changes were detected for TIM-3 or TIGIT. In CD4 ⁺ T cells, no significant modulation of IC molecules was observed, whereas IL-22 secretion was reduced. Conclusions: These findings support an immunomodulatory effect of 1,25(OH) ₂ vitamin D on human CD8 ⁺ T cells, with attenuation of effector-associated responses and modulation of checkpoint-associated pathways in vitro . Our results suggest that vitamin D may contribute to the regulation of inflammatory T-cell responses under controlled culture conditions. The functional and therapeutic implications of these observations require further investigation.

PDZ domains are involved in many cellular processes and are key regulators of the cell physiology. A huge number of studies have investigated the binding specificity of PDZ domains to the carboxyl-terminal sequence of target proteins, while the molecular mechanisms that mediate the recognition of internal binding regions are largely unexplored. In the present study, we describe a ligand motif located in the catalytic domain of the phosphatase Dusp26 which mediates its binding to the PDZ-4 of Scribble. Site-directed mutagenesis identified a conserved tyrosine residue as relevant for the binding. The interaction with the PDZ domain could help the phosphatase to recruit its specific targets.

Eukaryotic cells are intracellularly divided into several compartments that provide spatiotemporal control over biochemical reactions. Phase separation of proteins and RNA is emerging as an important mechanism underlying the formation of intracellular compartments that are not delimited by membranes. These structures are also known as biomolecular condensates and have been shown to serve a myriad of cellular functions, such as organization of cytoplasm and nucleoplasm, stress response, signal transduction, gene regulation, and immune response. Here, the author will summarize our current understanding of intracellular phase separation, its biological functions, and how this phenomenon is regulated in eukaryotic cells. Additionally, the author will review recent evidence of the role of biomolecular condensates in the development of pathophysiological conditions, with special emphasis on cancer and immune signaling.

People living with HIV (PLWH) represent a population of particular interest in the context of arboviral infections. This observational study aimed to evaluate and compare the magnitude and functionality of dengue virus (DENV)-specific T-cell memory elicited by natural DENV infection in PLWH and HIV-negative individuals. Nineteen participants infected during the 2023 autochthonous dengue outbreak in Rome (10 PLWH and 9 HIV-negative individuals) were assessed for interferon gamma (IFN-γ)-releasing cells in peripheral blood mononuclear cells (PBMCs) using an IFN-γ -ELISpot assay following stimulation with peptide pools.When examining both the overall cumulative ELISpot responses, calculated as the sum of spot-forming units (SFU) across all antigens or the binary responder status to each DENV antigen, HIV-negative individuals showed higher and broader cumulative responses than PLWH, but differences did not reach statistical significance. However, responses to prM/E/C antigens were higher in the HIV-negative group, reaching statistical significance ( P = 0.041). Among PLWH, no association was found between T-cell reactivity and CDC stage at diagnosis. Our findings suggest that while PLWH with well-controlled infection retain dengue-specific T-cell memory, their response to structural antigens may be attenuated. Vaccination should be considered a key preventive strategy for this clinically vulnerable population. IMPORTANCE: The geographic expansion of dengue virus (DENV) transmission into temperate regions highlights the need to characterize immune responses in populations with chronic conditions such as HIV. Despite immune reconstitution under antiretroviral therapy (ART), people living with HIV (PLWH) may exhibit residual immune dysregulation that could affect DENV-specific immunity and clinical outcomes. In this study, we directly compared DENV-specific T-cell memory responses in PLWH and HIV-negative individuals following natural infection during an autochthonous outbreak. While PLWH mounted detectable cellular responses, reactivity to structural antigens (prM/E/C) was significantly reduced compared with HIV-negative participants. These findings indicate that qualitative differences in antigen-specific T-cell memory may persist despite virological suppression and immune recovery, with potential implications for protection upon re-exposure. Our results provide a rationale for evaluating dengue vaccination strategies in PLWH as local transmission becomes more frequent in non-endemic settings.

Longitudinal T cell receptor repertoire analysis can confound biological change with measurement noise and estimator-induced structure. We developed ClonoDynamics, a framework integrating latent abundance inference with replicate-decoupled estimates of directional change and cross-replicate displacement covariance. Pseudo-longitudinal series assembled from 12 independently processed technical measurements of one biological sample revealed apparent restoring dynamics caused by reusing measurements for conditioning and change estimation. Separating these roles between replicates largely removed this artifact. Synthetic controls preserving empirical detection patterns showed dose-dependent recovery of temporal accumulation, resolving the two larger imposed effects but not the smallest. Applied to TCRβ repertoires from ten healthy adults sampled weekly for six weeks, ClonoDynamics identified abundance-dependent directional change and shared fluctuations exceeding the technical reference. Within the analyzed abundance domain, these fluctuations showed no consistent positive accumulation over 1–5 weeks. ClonoDynamics provides an experimentally benchmarked framework for distinguishing estimator-induced patterns, replicate-consistent longitudinal variation, and temporal accumulation. One Sentence Summary: ClonoDynamics combines technical replication and pseudo-longitudinal benchmarking to distinguish estimator-induced artifacts from replicate-consistent T cell receptor repertoire changes and assess their accumulation over time.