Phage display demonstrates durable differences in serological profile by route of inoculation in primary infections of non-human primates with Dengue Virus 1

Jayant V Rajan,Simon Pollett,Richard Jarman,Greg Gromowski,Caleigh Mandel-Brehm,Joseph L Derisi,Michael Mccracken

doi:10.1038/s41598-021-90318-z

Abstract

Natural dengue virus (DENV) infections occur by mosquito bite but how the inoculation route affects the humoral immune response is unknown. We serologically profiled 20 non-human primates (NHP) from a prior study of DENV1 infection where animals were inoculated by mosquito (N = 10) or subcutaneous injection (N = 10). Using a comprehensive, densely tiled and highly redundant pan-flavivirus programmable phage library containing 91,562 overlapping 62 amino acid peptides, we produced a high-resolution map of linear peptide sequences enriched during DENV seroconversion. Profiles in mosquito-inoculated and subcutaneously-inoculated animals were similar up to 90 days after primary infection, but diverged at 1 year with differences in sero-reactivity in the Envelope (E; residues 215–406; p < 0.08), and Nonstructural-3 (NS3; residues 549–615; p < 0.05) proteins in mosquito-inoculated versus subcutaneously-inoculated animals. Within the E protein, residues 339–384 in domain III accounted for > 99% of the observed sero-reactivity difference. Antibody breadth did not vary by mode of inoculation. The differential reactivity to E domain III seen by phage display validated orthogonally by ELISA, but did not correlate with late neutralization titers. Serological profiling of humoral immune responses to DENV infection in NHP by programmable phage display demonstrated durable differences in sero-reactivity by route of inoculation.

Highlights

Natural dengue virus (DENV) infections occur by mosquito bite but how the inoculation route affects the humoral immune response is unknown
The programmable phage library used here consists of 91,562 overlapping 62 amino acid peptides derived from 74 flaviviruses
For the 10 non-human primates (NHP) inoculated by subcutaneous injection of DENV1 the number of enriched peptides at [7, 35, 90] and 365 days p.i. ranged from 26 to [93, 88] to 930, 64 to 789 and 31 to 786

Summary

Introduction

Natural dengue virus (DENV) infections occur by mosquito bite but how the inoculation route affects the humoral immune response is unknown. We serologically profiled 20 non-human primates (NHP) from a prior study of DENV1 infection where animals were inoculated by mosquito (N = 10) or subcutaneous injection (N = 10). Serological profiling of humoral immune responses to DENV infection in NHP by programmable phage display demonstrated durable differences in sero-reactivity by route of inoculation. Much of the current understanding of DENV immunity in humans derives from studies of human sera as well as studies of monoclonal antibodies isolated from B-cells of persons infected with DENV. Such studies have focused on the analysis of natural DENV infections delivered by Aedes mosquitoes. Programmable phage display immunoprecipitation sequencing (PhIP-Seq), a high throughout serological profiling technique, was used to characterize serological responses to primary DENV1 infection as well as to both homologous (DENV1) and heterologous (DENV2) challenge in rhesus macaques

Objectives

Methods

Results

Conclusion