The 31.2% decrease in HIV acquisition reported in the RV144 trial has raised hopes that vaccine protection may be achievable. We have pursued a bimodal vaccine approach to induce both cellular and humoral immunity;in our recent rhesus macaque (RM) study, recombinant protein immunogens (SIV Gag-Pol particles, HIV Tat, and multimeric HIV clade C (HIV-C) gp160) provided complete protection for some RMs against multiple intrarectal challenges with a heterologous R5 clade C SHIV (SHIV-C). Our study simultaneously linked cellular as well as humoral antiviral immunity to protection. Overall, five vaccine-protected RMs remained free of persistent, systemic infection;all had generated high-titer neutralizing antibodies (nAbs) in response to multimeric gp160 of an HIV-C strain that diverged by 22.2% in amino acid sequence from Env of the challenge virus. Our overall hypothesis is that vaccine-induced Abs - either nAbs and/or Abs with antiviral effector functions - can protect against heterologous virus acquisition. We have developed new tools to determine the epitope specificity of protective Abs from polyclonal sera. Our strategy involves a) differentil biopanning with recombinant peptide libraries to isolate mimotopes linked to protection, b) isolation of single RM B cells specific for a given mimotope/epitope, 3) PCR amplification of RM immunoglobulin variable regions, and 4) generation of recombinant Abs. These novel approaches have led to the isolation of two new chimeric simian/human nmAbs with predicted epitope specificity. We now seek to use these new tools for the following Specific Aims: 1. to characterize the epitopes recognized by polyclonal Abs of vaccine-protected RMs by differential biopanning. First, we will positively select recombinant phages encoding random peptide libraries by biopanning with plasma from a vaccine-protected RM, followed by negative counter-selection with plasma from vaccinated, unprotected RMs. After several rounds of positive/negative selection, recombinant phages will reflect mimotopes linked to protection. We will also address the converse question: did RMs with vaccine failure mount unfavorable Ab responses that are not found in vaccine-protected RMs - or is failure simply a lack of protection-linked Abs? To do this, we will reverse the biopanning strategy and characterize the cognate epitopes linked to vaccine failure. 2. to isolate antigen-specific single B cells from the protecte RMs and PCR amplify the heavy/light chain variable immunoglobulin regions, using our newly generated RM-specific primers. 3. to perform passive immunization in RMs with the novel mAbs to demonstrate protection against mucosal challenge with a heterologous R5 SHIV. Our studies, which are based upon a well-characterized cohort of vaccine-protected RMs given upfront heterologous SHIV-C challenges, will identify epitopes that are protective or perhaps also deleterious in vivo and therefore provide important information for future HIV/AIDS immunogen design and optimization.

Public Health Relevance

This project seeks to analyze the differences in antibody responses induced by an AIDS vaccine between monkeys that were completely protected from AIDS virus infection and those that were not. We will isolate the B cells responsible for producing the successful antibodies, clone their antibody genes, and generate monoclonal antibodies, which will be tested for their specificity and potential to block AIDS virus infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI100703-02
Application #
8485541
Study Section
Special Emphasis Panel (ZRG1-AARR-E (02))
Program Officer
Li, Yen
Project Start
2012-06-08
Project End
2017-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
2
Fiscal Year
2013
Total Cost
$773,072
Indirect Cost
$279,662
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
076580745
City
Boston
State
MA
Country
United States
Zip Code
02215
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Scinto, Hanna B; Gupta, Sandeep; Thorat, Swati et al. (2018) Novel Strategy To Adapt Simian-Human Immunodeficiency Virus E1 Carrying env from an RV144 Volunteer to Rhesus Macaques: Coreceptor Switch and Final Recovery of a Pathogenic Virus with Exclusive R5 Tropism. J Virol 92:
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