Addressing <scp>HIV</scp>‐1 latency with <scp>Flow‐FISH</scp>: Finding, characterizing and targeting <scp>HIV</scp>‐1 infected cells

Heeren, Julian J Freen-van

doi:10.1002/cyto.a.24462

Cited by 5 publications

(9 citation statements)

References 18 publications

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“…HIV-1, a retrovirus, was described in 1983 as the causative agent of acquired immunodeficiency syndrome, or AIDS [65]. Since then, the diagnosis, treatment, and monitoring of AIDS patients has progressed massively [4,20,66,67]. With the advent of antiretroviral therapy, HIV-1 propagation can be kept in check, preventing the development of AIDS.…”

Section: Hiv-1mentioning

confidence: 99%

“…Indeed, HIV-1 Flow-FISH assays have been employed to study infected cell lines [69], and ex vivo infected cells, such as total PBMCs [70], epidermal DCs [71], and T cells [72,73], but also patient-derived material, such as alveolar macrophages [74], T cells [3,19,74,75], and even platelets [75]. In the future, Flow-FISH might even be useful as a tool to monitor therapy effectiveness and assessment of total eradication of (latently) infected cells [20].…”

Section: Hiv-1mentioning

confidence: 99%

“…Latency reversal [81] Cell lines Ex vivo infected T cells Latency reversal [82] Flow-FISH analysis can also potentially lead to new therapeutic insights [20]. For instance, Flow-FISH was used to show that patient-derived HIV-1-infected T cells expressed CD20 upon viral reactivation [77].…”

Section: Cell Lines Patient T Cellsmentioning

confidence: 99%

“…As discussed, if properly treated with antiretroviral therapy, HIV-1 forms a latent infection cycle. Identifying and characterizing the latently infected cells and, perhaps even more importantly, the cells forming the translationally-competent and replication-competent reservoir could lead to new therapeutic strategies [20]. Firstly, identifying cells that form the latent reservoir could be instrumental for the eradication of these specific viral reservoirs.…”

Section: Cell Lines Patient T Cellsmentioning

confidence: 99%

“…Flow-FISH has mainly been employed in three fields of study: (1) studying T cell biology and effector function [2,[12][13][14], (2) assessing telomere length [15,16], and (3) detecting and studying (intracellular) bacteria, fungi, and viruses [17][18][19][20]. Here, the use of Flow-FISH for the detection of (intracellular) pathogens, studying microorganism biology and disease pathogenesis, and the identification of bacterial, fungal, and viral strains in mixed cultures is discussed, with a particular focus on studies investigating Epstein-Barr virus (EBV), human immunodeficiency virus 1 (HIV-1), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent responsible for the coronavirus disease 2019 (COVID-19) pandemic.…”

Section: Introductionmentioning

confidence: 99%

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Flow-FISH as a Tool for Studying Bacteria, Fungi and Viruses

Heeren¹

2021

BioTech

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Many techniques are currently in use to study microbes. These can be aimed at detecting, identifying, and characterizing bacterial, fungal, and viral species. One technique that is suitable for high-throughput analysis is flow cytometry-based fluorescence in situ hybridization, or Flow-FISH. This technique employs (fluorescently labeled) probes directed against DNA or (m)RNA, for instance targeting a gene or microorganism of interest and provides information on a single-cell level. Furthermore, by combining Flow-FISH with antibody-based protein detection, proteins of interest can be measured simultaneously with genetic material. Additionally, depending on the type of Flow-FISH assay, Flow-FISH can also be multiplexed, allowing for the simultaneous measurement of multiple gene targets and/or microorganisms. Together, this allows for, e.g., single-cell gene expression analysis or identification of (sub)strains in mixed cultures. Flow-FISH has been used in mammalian cells but has also been extensively employed to study diverse microbial species. Here, the use of Flow-FISH for studying microorganisms is reviewed. Specifically, the detection of (intracellular) pathogens, studying microorganism biology and disease pathogenesis, and identification of bacterial, fungal, and viral strains in mixed cultures is discussed, with a particular focus on the viruses EBV, HIV-1, and SARS-CoV-2.

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Section: Hiv-1mentioning

confidence: 99%

Section: Hiv-1mentioning

confidence: 99%

Section: Cell Lines Patient T Cellsmentioning

confidence: 99%

Section: Cell Lines Patient T Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Flow-FISH as a Tool for Studying Bacteria, Fungi and Viruses

Heeren¹

2021

BioTech

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FISHing for COVID‐19: Studying SARS‐CoV‐2 with single‐molecule FISH and flow‐FISH

Heeren¹

2022

Cytometry Pt A

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For more than 2 years, the COVID-19 pandemic has plagued the world. The causative agent, SARS-CoV-2, is a single-stranded RNA virus that is easily transmitted, resulting in >603 million confirmed infections worldwide (https://covid19.who.int/) at the time of writing.Many efforts have been undertaken to unravel SARS-CoV-2 biology, epidemiology and pathology, but also the immune responses directed against the virus has been thoroughly investigated. To do so, new reagents were required to study this newly emerged virus, and novel assays were developed, ranging from diagnostic PCR tests, antibody ELISAs and at-home test kits, all with the aim to study and combat the COVID-19 pandemic.However, many of these assays make use of, or detect, isolated viral genetic material or viral proteins, thus not allowing to study infected cells at a single cell level. One of the assays that has been developed and allows for studying SARS-CoV-2 infection at the single cell level makes use of single-molecule fluorescence in situ hybridization (smFISH) [1]. By employing probe sets that target the conserved regions of the SARS-CoV-2 genome, viral RNA can be visualized at the single molecule level. Authors showed that in Vero cells, a primate cell-line, the FISH assay was suitable to detect the level of SARS-CoV-2 infection. Also human cell lines, human post-mortem patient

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HIV infection detection using CRISPR/Cas systems: Present and future prospects

Deng,

Xue

2023

Computational and Structural Biotechnology Journal

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Addressing HIV‐1 latency with Flow‐FISH: Finding, characterizing and targeting HIV‐1 infected cells

Cited by 5 publications

References 18 publications

Flow-FISH as a Tool for Studying Bacteria, Fungi and Viruses

Flow-FISH as a Tool for Studying Bacteria, Fungi and Viruses

FISHing for COVID‐19: Studying SARS‐CoV‐2 with single‐molecule FISH and flow‐FISH

HIV infection detection using CRISPR/Cas systems: Present and future prospects

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