# Optimization of high throughput viral outgrowth assays for the detection of HIV-1 reservoirs

> **NIH NIH R01** · JOHNS HOPKINS UNIVERSITY · 2020 · $409,375

## Abstract

Project Summary
 Latently infected CD4+ T cells represent the major barrier to HIV-1 eradication.
Many strategies are being developed to eliminate these cells, but the field lacks a simple
sensitive assay that can screen for these cells. PCR based assays cannot distinguish
between replication-competent versus defective virus and recent studies have shown
that the current state of the art culture assay measures only a small fraction of the
replication-competent virus present. In this proposal we aim to develop a sensitive assay
that is capable of selectively amplifying replication-competent virus from latently infected
CD4+ T cells. Such an assay could inform the decision of whether or not to discontinue
cART in patients after a therapeutic intervention is made. The need for such an assay is
highlighted by the recent report of the 2 “Boston patients” who were treated with
allogeneic stem cell transplantation for malignancies. No HIV DNA was detected by PCR
in either patient from as many as 200 million PBMCs (<0.07 copies/106 PBMCs), but
after cART was discontinued there was a rapid rebound in viremia in both patients. This
outcome is undesirable is it negates the theoretical advantages to having smaller HIV-1
reservoirs. The development of a simple but sensitive assay that can screen a very large
number of CD4+ T cells may help prevent the occurrence of a similar scenario from
occurring in the future.
 We will use the optimized assay we develop to screen CD4+ T cells from patients
who were treated with ART during primary infection and correlate the number of latently
infected cells as measured in our assay to the time it took for viral rebound to occur
when ART was discontinued in these subjects.
 This work will be important for the development of strategies for HIV-1 eradication
since the assay will help inform the decision of whether or not to discontinue
antiretroviral therapy.

## Key facts

- **NIH application ID:** 9949632
- **Project number:** 5R01AI140789-03
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** JOEL N BLANKSON
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $409,375
- **Award type:** 5
- **Project period:** 2018-07-05 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9949632

## Citation

> US National Institutes of Health, RePORTER application 9949632, Optimization of high throughput viral outgrowth assays for the detection of HIV-1 reservoirs (5R01AI140789-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9949632. Licensed CC0.

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