During the first year of the EBV-MS project, the consortium advanced several key scientific and technical components across clinical, immunological, genomic, epidemiological, and computational domains:
Clinical Trials: The phase 2a clinical trial investigating the antiviral drug tenofovir alafenamide fumarate (TAF) in MS patients was initiated, with patient recruitment and randomization ongoing across multiple centers. The trial includes comprehensive clinical assessments, MRI imaging, and frequent saliva and blood sampling for EBV and immunological analyses. Preparations for the phase 2b trial were completed, including protocol development and integration of advanced biomarkers such as neurofilament light (NFL) in cerebrospinal fluid, MRI-based inflammation markers, and PET imaging.
Immunological Profiling: Extensive protocols were developed and implemented for flow cytometry, antigen stimulation, and multiomic analyses. Blood samples from MS patients treated with anti-CD20 therapies were collected and processed to assess immune cell activation and antigen-specific responses. Single-cell RNA sequencing and methylation profiling were performed on sorted memory B and T cells, and immunophenotyping of circulating immune cells was initiated using spectral cytometry.
EBV Genomics: Saliva and blood samples were collected from MS patients and controls across several European centers. DNA extraction and EBV genome enrichment protocols were optimized, and sequencing workflows were established. A bioinformatics pipeline was developed for processing viral sequences and identifying genetic variants, enabling future analyses of EBV-host interactions and viral genome-wide association studies.
Epidemiological Analysis: Applications for access to national health registries were submitted and approved to enable linkage of EBV infection data with MS onset and progression. Coordination among partners was established to prepare for analyses of the timing between EBV infection and MS onset, and to identify drugs that may influence this interval.
Computational Modeling: A mathematical model of the immune system was developed using ordinary differential equations to simulate interactions between effector and regulatory T cells, B cells, and natural killer cells in the context of EBV infection. The model supports in silico testing of hypotheses such as molecular mimicry, mistaken self, and impaired immune regulation. Simulations were conducted to explore immune dynamics from healthy states through EBV infection to MS-like inflammatory relapses. A complementary model of CNS damage is under development for integration with the immune system model.
Data Management and Integration: A comprehensive data management plan was implemented, including the development of a data dictionary and harmonization of clinical trial variables. Preparations were made for secure and interoperable data sharing across work packages, supporting future analyses and integration of clinical, immunological, and genomic data.
