NDLI: A passive-flow microfluidic device for imaging latent HIV activation dynamics in single T cells

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Integrative Biology Vol -9

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Patients are a virtue: advances in microengineered systems for clinical applications

Extracting physical chemistry from mechanics: a new approach to investigate DNA interactions with drugs and proteins in single molecule experiments

Agent-based model of angiogenesis simulates capillary sprout initiation in multicellular networks

A passive-flow microfluidic device for imaging latent HIV activation dynamics in single T cells

Extracellular matrix stiffness modulates VEGF calcium signaling in endothelial cells: individual cell and population analysis

Automated, high-throughput serum glycoprofiling platform

Cell spreading area regulates clathrin-coated pit dynamics on micropatterned substrate

Intrinsic and induced drug resistance mechanisms: in silico investigations at the cellular and tissue scales

Measurement of the entrainment window of islets of Langerhans by microfluidic delivery of a chirped glucose waveform

Printing cancer cells into intact microvascular networks: a model for investigating cancer cell dynamics during angiogenesis

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A passive-flow microfluidic device for imaging latent HIV activation dynamics in single T cells

Content Provider	Royal Society of Chemistry (RSC)
Author	Gearhart, Larisa M. Chavali, Arvind K. Miller-Jensen, Kathryn Ramji, Ramesh Wong, Victor C.
Copyright Year	2015
Abstract	Quantifying cell-to-cell variability in drug response dynamics is important when evaluating therapeutic efficacy. For example, optimizing latency reversing agents (LRAs) for use in a clinical “activate-and-kill” strategy to purge the latent HIV reservoir in patients requires minimizing heterogeneous viral activation dynamics. To evaluate how heterogeneity in latent HIV activation varies across a range of LRAs, we tracked drug-induced response dynamics in single cells via live-cell imaging using a latent HIV–GFP reporter virus in a clonal Jurkat T cell line. To enable these studies in suspension cells, we designed a simple method to capture an array of single Jurkat T cells using a passive-flow microfluidic device. Our device, which does not require external pumps or tubing, can trap hundreds of cells within minutes with a high retention rate over 12 hours of imaging. Using this device, we quantified heterogeneity in viral activation stimulated by transcription factor (TF) activators and histone deacetylase (HDAC) inhibitors. Generally, TF activators resulted in both faster onset of viral activation and faster rates of production, while HDAC inhibitors resulted in more uniform onset times, but more heterogeneous rates of production. Finally, we demonstrated that while onset time of viral gene expression and rate of viral production together predict total HIV activation, rate and onset time were not correlated within the same individual cell, suggesting that these features are regulated independently. Overall, our results reveal drug-specific patterns of noisy HIV activation dynamics not previously identified in static single-cell assays, which may require consideration for the most effective activate-and-kill regime.
Starting Page	998
Ending Page	1010
Page Count	13
File Format	HTM / HTML PDF
ISSN	17579694
Volume Number	7
Issue Number	9
Journal	Integrative Biology
DOI	10.1039/c5ib00094g
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	GFP HDAC HIV Petroleum reservoir Reporter gene Virus T cell Transcription factor Histone deacetylase Gene
Content Type	Text
Resource Type	Article
Subject	Medicine Biochemistry Biophysics

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Latent HIV in primary T lymphocytes is unresponsive to histone deacetylase inhibitors

Synergistic activation of HIV-1 expression by deacetylase inhibitors and prostratin: implications for treatment of latent infection (2009)

Histone Deacetylase Inhibitors for Purging HIV-1 from the Latent Reservoir

Development of an HIV reporter virus that identifies latently infected CD4⁺ T cells.

Runx1 Phosphorylation by Src Increases Trans-activation via Augmented Stability, Reduced Histone Deacetylase (HDAC) Binding, and Increased DNA Affinity, and Activated Runx1 Favors Granulopoiesis

Histone deacetylase inhibitors induce complex host responses that contribute to differential potencies of these compounds in HIV reactivation

Histone Deacetylase 3, a Class I Histone Deacetylase, Suppresses MAPK11-Mediated Activating Transcription Factor-2 Activation and Represses TNF Gene Expression

TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4⁺ T Cells.

Repression by HDAC 3 ( HDAC ) Inhibitor Trichostatin A and Factor 11 Gene by the Histone Deacetylase Activation of the Growth-Differentiation

A passive-flow microfluidic device for imaging latent HIV activation dynamics in single T cells

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Latent HIV in primary T lymphocytes is unresponsive to histone deacetylase inhibitors

Synergistic activation of HIV-1 expression by deacetylase inhibitors and prostratin: implications for treatment of latent infection (2009)

Histone Deacetylase Inhibitors for Purging HIV-1 from the Latent Reservoir

Development of an HIV reporter virus that identifies latently infected CD4+ T cells.

Runx1 Phosphorylation by Src Increases Trans-activation via Augmented Stability, Reduced Histone Deacetylase (HDAC) Binding, and Increased DNA Affinity, and Activated Runx1 Favors Granulopoiesis

Histone deacetylase inhibitors induce complex host responses that contribute to differential potencies of these compounds in HIV reactivation

Histone Deacetylase 3, a Class I Histone Deacetylase, Suppresses MAPK11-Mediated Activating Transcription Factor-2 Activation and Represses TNF Gene Expression

TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4+ T Cells.

Repression by HDAC 3 ( HDAC ) Inhibitor Trichostatin A and Factor 11 Gene by the Histone Deacetylase Activation of the Growth-Differentiation

A passive-flow microfluidic device for imaging latent HIV activation dynamics in single T cells

Development of an HIV reporter virus that identifies latently infected CD4⁺ T cells.

TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4⁺ T Cells.