Genotypic Analysis of HIV‐1 Drug Resistance at the Limit of Detection: Virus Production without Evolution in Treated Adults with Undetectable HIV Loads

Tara L. Kieffer; Mariel M. Finucane; Richard E. Nettles; Thomas C. Quinn; Karl W. Broman; Stuart C. Ray; Deborah Persaud; Robert F. Siliciano

doi:10.1086/382488

Natural Polymorphisms in the Human Immunodeficiency Virus Type 2 Protease Can Accelerate Time to Development of Resistance to Protease Inhibitors

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00870-06 ◽

2006 ◽

Vol 51 (2) ◽

pp. 604-610 ◽

Cited By ~ 43

Author(s):

Michel Ntemgwa ◽

Bluma G. Brenner ◽

Maureen Oliveira ◽

Daniela Moisi ◽

Mark A. Wainberg

Keyword(s):

Human Immunodeficiency Virus ◽

Drug Resistance ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Serial Passage ◽

Drug Susceptibility ◽

Genotypic Analysis ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Human immunodeficiency virus type 2 (HIV-2) contains numerous natural polymorphisms in its protease (PR) gene that are implicated in drug resistance in the case of HIV-1. This study evaluated emergent PR resistance in HIV-2. Three HIV-2 isolates were selected for resistance to amprenavir (APV), nelfinavir (NFV), indinavir (IDV), and tipranavir (TPV) in cell culture. Genotypic analysis determined the time to the appearance of protease inhibitor (PI)-associated mutations compared to HIV-1. Phenotypic drug susceptibility assays were used to determine the levels of drug resistance. Within 10 to 15 weeks of serial passage, three major mutations—I54M, I82F, and L90M—arose in HIV-2 viral cultures exposed to APV, NFV, and IDV, whereas I82L was selected with TPV. After 25 weeks, other cultures had developed I50V and I84V mutations. In contrast, no major PI mutations were selected in HIV-1 over this period except for D30N in the context of NFV selective pressure. The baseline phenotypes of wild-type HIV-2 isolates were in the range observed for HIV-1, except for APV and NFV for which a lower degree of sensitivity was seen. The acquisition of the I54M, I84V, L90M, and L99F mutations resulted in multi-PI-resistant viruses, conferring 10-fold to more than 100-fold resistance. Of note, we observed a 62A/99F mutational motif that conferred high-level resistance to PIs, as well as novel secondary mutations, including 6F, 12A, and 21K. Thus, natural polymorphisms in HIV-2 may facilitate the selection of PI resistance. The increasing incidence of such polymorphisms in drug-naive HIV-1- and HIV-2-infected persons is of concern.

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The CNS in the face of ART contains T cell origin HIV which can lead to drug resistance

10.1101/588426 ◽

2019 ◽

Cited By ~ 2

Author(s):

Gila Lustig ◽

Sandile Cele ◽

Farina Karim ◽

Yashica Ganga ◽

Khadija Khan ◽

...

Keyword(s):

Drug Resistance ◽

T Cell ◽

Limit Of Detection ◽

Virus Production ◽

Surface Markers ◽

Therapy Failure ◽

Significant Difference ◽

Cellular Source ◽

Csf Escape

AbstractHIV persists despite antiretroviral therapy (ART) in cellular reservoirs thought to occur in distinct anatomical compartments. Therapy failure may occur because of incomplete ART adherence and possibly viral replication at some reservoir sites. The CNS may serve as a reservoir site due to lowered ART penetration and virus production from long-lived tissue resident macrophages. Compelling evidence for the CNS as a reservoir is the existence of individuals where HIV is suppressed below limit of detection in blood but detectable in the cerebrospinal fluid (CSF), termed CSF Escape. Here, we asked whether HIV in CSF Escape individuals is derived from macrophages or persists due to lowered ART. We used cell surface markers on the HIV envelope to determine the cellular source of HIV. We verified detection usingin vitroderived virus from infected macrophages and T cells and tested CSF from CSF Escape individuals. We observed host surface markers consistent with T cell origin. We also measured ART concentrations in the CSF and plasma. We found a dramatic decrease in CSF ART concentrations described previously, but no significant difference between CSF Escape versus fully suppressed individuals. To examine the effect of the observed CSF ART concentrations on HIV replication, we used long-term infection with ART in cell culture. CSF Escape ART levels led to either HIV suppression or evolution of drug resistance, but not replication of drug sensitive HIV. These observations argue that persistent CNS viremia despite ART can be T cell generated and may result in drug resistance and therapy failure.

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Comparison of in-house HIV-1 genotypic drug resistant test with commercial HIV-1 genotypic test kit

Asian Biomedicine ◽

10.5372/1905-7415.0502.032 ◽

2011 ◽

Vol 5 (2) ◽

pp. 249-255 ◽

Cited By ~ 1

Author(s):

Jutarat Praparattanapan ◽

Yingmanee Tragoolpua ◽

Jeerang Wongtrakul ◽

Wilai Kotarathitithum ◽

Romanee Chaiwarith ◽

...

Keyword(s):

Drug Resistance ◽

Limit Of Detection ◽

Standard Of Care ◽

Resource Limited Settings ◽

Genotypic Resistance ◽

Resource Limited ◽

Number Of Patients ◽

Test Kit ◽

Commercial Kits ◽

Hiv 1

Abstract Background: The use of combination antiretroviral therapy (cART) has become a standard of care in the treatment of HIV infection. However, antiretroviral drug resistance occurs in a substantial number of patients. In resource-limited settings, genotypic resistance assay using a commercial kit is costly. Objective: Focus on the validation of an in-house HIV-1 specific genotypic drug resistance assay in Thai patients failing cART. Materials and methods: Results of HIV-1 genotypic drug resistance assay was evaluated by comparing an inhouse method to a commercial test. The TRUGENE HIV-1 genotyping kit was used in 79 plasma specimens (49 from HIV patients failing cART therapy and 30 from proficiency testing panels). Results: The results from the in-house assay were comparable to those obtained from the TRUGENE HIV-1 genotyping kit with >99.0% codon-to-codon agreement. The lower limit of detection by the in-house assay was approximately 100 copies/mL of HIV-1 RNA. In addition, this in-house assay would allow testing of samples from patients infected with HIV-1 subtype other than B. Conclusion: The in-house HIV-1 genotypic drug resistance assay may be used as an alternative to commercial kits, particularly in resource limited settings.

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Ultrastructural observations of human monocytes infected with HIV-1

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100084843 ◽

1991 ◽

Vol 49 ◽

pp. 108-109

Author(s):

James K. Koehler ◽

Steven G. Reed ◽

Joao S. Silva

Keyword(s):

Gradient Centrifugation ◽

Virus Production ◽

Human Monocyte ◽

Red Cross ◽

Human Monocytes ◽

Blood Monocytes ◽

Hiv Replication ◽

Electron Microscopic ◽

Ultrastructural Studies ◽

Hiv 1

As part of a larger study involving the co-infection of human monocyte cultures with HIV and protozoan parasites, electron microscopic observations were made on the course of HIV replication and infection in these cells. Although several ultrastructural studies of the cytopathology associated with HIV infection have appeared, few studies have shown the details of virus production in “normal,” human monocytes/macrophages, one of the natural targets of the virus, and suspected of being a locus of quiescent virus during its long latent period. In this report, we detail some of the interactions of developing virons with the membranes and organelles of the monocyte host.Peripheral blood monocytes were prepared from buffy coats (Portland Red Cross) by Percoll gradient centrifugation, followed by adherence to cover slips. 90-95% pure monocytes were cultured in RPMI with 5% non-activated human AB serum for four days and infected with 100 TCID50/ml of HIV-1 for four hours, washed and incubated in fresh medium for 14 days.

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