Novel Single-Cell-Level Phenotypic Assay for Residual Drug Susceptibility and Reduced Replication Capacity of Drug-Resistant Human Immunodeficiency Virus Type 1

• DOI: 10.1128/jvi.78.4.1718-1729.2004
• Publication Date: 2004-01-27T19:58:01Z
• Authors: Haili Zhang; Yan Zhou; Cecily Alcock; Tara Kiefer; Daphne Monie; Janet Siliciano; Quan Li; Paul Pham; Joseph Cofrancesco; Deborah Persaud; Robert F. Siliciano
• Keywords: Antiviral drug

Human immunodeficiency virus type 1 (HIV-1)-infected individuals who develop drug-resistant during antiretroviral therapy may derive benefit from continued treatment for two reasons. First, viruses can retain partial susceptibility to the drug combination. Second, selects that have reduced replication capacities relative archived, drug-sensitive viruses. We developed a novel single-cell-level phenotypic assay allows these effects be distinguished and compared quantitatively. Patient-derived gag-pol sequences were cloned into an HIV-1 reporter expresses endoplasmic reticulum-retained Env-green fluorescent protein fusion. Flow cytometric analysis of single-round infections allowed quantitative viral over 4-log dynamic range. The faithfully reproduced known in vivo interactions occurring at level target cells. Simultaneous by wild-type resistant presence absence relevant combination divided nonsuppressive additive components, residual selection variants with diminished capacities. In some patients resistance, dominant circulating retained significant However, other cases, showed no but had capacity virus. this case, simplification regimen might still allow adequate suppression third pattern, nevertheless equivalent such there is treatment. Thus, ability simultaneously analyze provide basis rational therapeutic decisions setting failure.