Core B: Biosafety Level 4 Core

BSL-4 CORE (Core B): PROJECT SUMMARY/ABSTRACT The pathogenesis of Ebola virus (EBOV) disease is characterized by a dysregulated immune response that includes the suppression of the innate immune system leading to "immune paralysis" and paradoxically activation of inflammatory pathways characteristic of a "cytokine storm". The goal of this Program Project is to investigate the molecular mechanisms of the dysregulated immune response to EBOV infections. EBOV infections must be performed under Biosafety Level 4/Animal Biosafety level 4 (BSL-4/ABSL-4) containment in accordance with government and institutional biosafety, biosecurity and select agent regulations; therefore, this Program Project requires a BSL-4 Core (Core B). Core B will use the BSL-4 and ABSL-4 facilities of the Galveston National Laboratory (GNL), which is part of the University of Texas Medical Branch (UTMB). Interactions between Core B and the Research Projects (RPs) will be coordinated by the Administrative Core (Core A). Fundamental to the efforts included in the Program Project will be performing infections of primary immune and nonimmune cells from deidentified human subjects with EBOV and a mutant with a disabled interferon-inhibiting domain. Next, cells and tissues will be taken from rhesus macaques infected with the same viruses, which will be used to complement in vitro data. The infected cells will be used for investigation of transcriptional (Research Project [RP]1), posttranscriptional (RP2), and posttranslational mechanisms (RP3) that lead to the dysregulated immune response to EBOV. The initial steps of the experimental procedures, which involve infectious materials, for all RPs and targeting of critical nodes leading to dysregulated responses at gene expression levels in vitro will be performed in GNL BSL-4 facilities. The in vivo validation of the targeting will be performed in mice and rhesus macaques will be performed in GNL ABSL-4 facilities. The procedures that will be performed in Core B are critical for achieving the major goals of the proposed studies, including: (a) detailed knowledge of transcriptional mechanisms leading to the dysregulated immune response to EBOV, (b) elucidation of the role of posttranscriptional mechanisms in the dysregulated immune response to EBOV, (c) elucidation of the role of posttranslational mechanisms in the dysregulated immune response to EBOV, and (d) building and experimental validation of a comprehensive model for the pathogenesis of EBOV infection and other severe viral infections, such as Marburg, Lassa fever, dengue, and COVID-19. Completion of Core B Specific Aims will provide critical support for RP1, RP2, and RP3 to identify the role of transcriptional, posttranscriptional, and posttranslational mechanisms in the dysregulated immune response to EBOV.