Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1.
The gp41 membrane-proximal external region (MPER) is a target for broadly neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). However, replication-defective virus vaccines currently under evaluation in clinical trials do not efficiently elicit MPER-specific antibodies. Structural modeling suggests that the MPER forms an α-helical coiled coil that is required for function and immunogenicity. To maintain the native MPER conformation, we used reverse genetics to engineer replication-competent reovirus vectors that displayed MPER sequences in the α-helical coiled-coil tail domain of viral attachment protein σ1. Sequences in reovirus strain type 1 Lang (T1L) σ1 were exchanged with sequences encoding HIV-1 strain Ba-L MPER epitope 2F5 or the entire MPER. Individual 2F5 or MPER substitutions were introduced at virion-proximal or virion-distal sites in the σ1 tail. Recombinant reoviruses containing heterologous HIV-1 sequences were viable and produced progeny yields comparable to those with wild-type virus. HIV-1 sequences were retained following 10 serial passages in cell culture, indicating that the substitutions were genetically stable. Recombinant viruses engineered to display the 2F5 epitope or full-length MPER in σ1 were recognized by purified 2F5 antibody. Inoculation of mice with 2F5-containing vectors or rabbits with 2F5- or MPER-containing vectors elicited anti-reovirus antibodies, but HIV-1-specific antibodies were not detected. Together, these findings indicate that heterologous sequences that form α-helices can functionally replace native sequences in the α-helical tail domain of reovirus attachment protein σ1. However, although these vectors retain native antigenicity, they were not immunogenic, illustrating the difficulty of experimentally inducing immune responses to this essential region of HIV-1. IMPORTANCE Vaccines to protect against HIV-1, the causative agent of AIDS, are not approved for use. Antibodies that neutralize genetically diverse strains of HIV-1 bind to discrete regions of the envelope glycoproteins, including the gp41 MPER. We engineered recombinant reoviruses that displayed MPER epitopes in attachment protein σ1 (REO-MPER vectors). The REO-MPER vectors replicated with wild-type efficiency, were genetically stable, and retained native antigenicity. However, we did not detect HIV-1-specific immune responses following inoculation of the REO-MPER vectors into small animals. This work provides proof of principle for engineering reovirus to express antigenic epitopes and illustrates the difficulty in eliciting MPER-specific immune responses.
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Karl W
KW
Boehme
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Mine'
M
Ikizler
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Jason A
JA
Iskarpatyoti
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
J Denise
JD
Wetzel
Department of Chemistry, Vanderbilt University, Nashville, Tennessee, USA; Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Jordan
J
Willis
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
James E
JE
Crowe
Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA; Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA; Laboratory for AIDS Vaccine Research and Development, Duke University Medical Center, Durham, North Carolina, USA.
Celia C
CC
LaBranche
Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA; Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA; Laboratory for AIDS Vaccine Research and Development, Duke University Medical Center, Durham, North Carolina, USA.
David C
DC
Montefiori
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Gregory J
GJ
Wilson
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Terence S
TS
Dermody
human immunodeficiency virus; immunization; live vector vaccines; neutralizing antibodies; reovirus