# PROJECT 4: Genetic and Structural Basis for Human Antibody Inhibition of Dengue Viruses

> **NIH NIH P01** · UNIVERSITY OF CALIFORNIA BERKELEY · 2021 · $400,751

## Abstract

PROJECT 4: Genetic and Structural Basis for Human Antibody Inhibition of Dengue Viruses
(Vanderbilt University)
SUMMARY
One of the primary goals of this P01 program project proposal is to define the functional and antigenic specificity
of the memory humoral response before and after dengue virus (DENV) infection. Here, we will determine the
molecular and structural basis for type-specific and cross-reactive human B cell responses to DENV infection.
Aim 1 of Project 4 will study the epitopes and structures recognized by DENV3 type-specific potently neutralizing
antibodies (Abs). The goal is to define a comprehensive antigenic map of neutralizing determinants on the DENV
E protein, which will inform DENV vaccine design and testing efforts. In past work with investigators in this
consortium, we have mapped many of the major antigenic determinants on DENV1 and DENV2. The protective
determinants on DENV3 are less well studied. Fortunately, in preliminary experiments, in collaboration with
Project 1 and Core C, we have generated a significant panel of potent DENV3-specific neutralizing monoclonal
Abs (mAbs) from the Nicaraguan Pediatric Dengue Cohort Study (PDCS) samples. It is clear that many of these
antibodies recognize novel epitopes that are not previously known. We will use these reagents to identify the
molecular and structural basis for recognition using alanine scanning mutagenesis, hydrogen deuterium
exchange mass spectrometry, cryo-EM and crystallography studies. If the antigenic maps appear incomplete,
as determined by Project 1 in dengue-endemic populations, we will generate additional mAbs from the peripheral
blood mononuclear cells (PBMCs) collected from children with documented repeat DENV3 infections, as well as
DENV1 and DENV2 infections, in the PDCS to define the complete antigenic landscape. We hypothesize that
most DENV3-specific Abs are directed to quaternary epitopes, including those near the DENV envelope domain
I/II (EDI/II) hinge region, but that there is a diversity of binding poses and angles for recognition of this complex
region. In Aim 2, we will define comprehensive antigenic maps for cross-reactive Abs that recognize and
neutralize viruses of all 4 DENV serotypes. In preliminary studies, we have isolated broad and potent mAbs that
have distinct profiles from those of the limited number of E dimer epitope (EDE) mAbs reported to date. The new
mAbs suggest there are additional sites of vulnerability for broad and potent neutralizing responses that are not
yet understood. Studies in this aim will define with biochemical, genetic and structural approaches the novel
epitopes associated with highly neutralizing cross-reactive mAbs that differ from EDE epitopes after 2° DENV
infection. Finally, in Aim 3, we will use emerging techniques in adaptive immune receptor next generation
sequencing to interrogate the Ab variable gene repertoires in PDCS subjects. We will use deep sequencing of
peripheral blood Ab gene repertoires in pri...

## Key facts

- **NIH application ID:** 10244879
- **Project number:** 5P01AI106695-07
- **Recipient organization:** UNIVERSITY OF CALIFORNIA BERKELEY
- **Principal Investigator:** James E Crowe
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $400,751
- **Award type:** 5
- **Project period:** 2015-07-29 → 2025-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10244879

## Citation

> US National Institutes of Health, RePORTER application 10244879, PROJECT 4: Genetic and Structural Basis for Human Antibody Inhibition of Dengue Viruses (5P01AI106695-07). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10244879. Licensed CC0.

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