The Administrative Core A, led by Dr. Richard Wyatt, will administer support for Projects 1 and 2 and Cores B and C, under the umbrella of this new HIVRAD proposal. This Core will coordinate an administrative partnership plan to maintain good communications within the HIVRAD working group and relevant external researchers, advisors and stakeholders related to design of novel cleavage-independent trimers and the analysis of B cell responses generated by vaccination of these HIV-derived envelope glycoprotein (Env) timers into non-human primates (NHPs). The Administrative Core will establish a HIVRAD website as well as cloud data storage accessible to all component PIs or/and designates. A major scientific objective, to elicit and define tier 2 cross- neutralizing B cell and antibody responses (bNAbs) in NHPs following vaccination of novel Env trimers, will be forwarded by Core A. The purpose of Core A is embodied by the successful competition of the following tasks. We will perform project management, while implementing a developed plan to ensure the success of the overall P01 program. This approach will be accomplished by the Program Director, enhanced by real-world learning processes in the accomplished previous Wyatt.Scripps HIVRAD. In Project 2, the Core A will foster studies to better define the expressed NHP heavy and light chain repertoire to determine lineages and the levels of somatic hypermutation elicited by heterologous trimer prime:boost immunization in NHPs. We will examine responses elicited by the novel uncleaved near-native NFL trimers in functional and structural detail to identify neutralizing Abs for genetic analysis. We will use NGS analysis to follow Ab lineages to define affinity maturation, as well as to investigate if the Abs are archived in long-lived compartments (bone marrow). We will apply deep sequencing computational analysis of vaccine-induced Env-specific B cell responses to define populations of epitope-specific Abs in NHPs. We will investigate the evolution of Env-specific Ab lineages following immunization using the NGS-based tracing module within IgDiscover. We will define the maturation pathways of mAbs and use this information to re-design trimer immunogens to drive the responses in broadly neutralizing directions. We will use structural analysis of Env-specific serum Abs (Core C), and isolated mAbs from vaccinated NHPs (Project 2) for iterative trimer redesign. The HIVRAD research team will leverage our previous finding to advances the field's understanding of vaccine-elicited B cell responses to neutralizing determinants of HIV-1 in small animals and NHPs to a level not previously approachable. The continued development of novel B cell sorting probes (ordered cleavage-independent, Avi-tagged NFL trimers), advances in cloning methods, Ab expression, B cell germinal center and lymph node analytical capacities, Ab germline gene identification and Ab lineage tracing through NGS make this an extremely timely and important application. In this new application we will continue ?move the bar? forward towards the elicitation of bAbs by subunit Env vaccination using novel, covalent particulate display as well as the exciting recent advances in lipid nanoparticle mRNA expression of cell-surface tethered NFL trimers.
