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

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

Content Provider	Europe PMC
Author	Sanyal, Anwesha Rangachar, Vatsala S Gupta, Phalguni
Copyright Year	2019
Abstract	The latent HIV-1 viral reservoir in resting CD4+ (rCD4+) T cells represents a major barrier to an HIV-1 cure. There is an ongoing effort to identify therapeutic approaches that will eliminate or reduce the size of this reservoir. However, clinical investigators lack an assay to determine whether or not a decrease in the latent reservoir has been achieved. Therefore, it is critical to develop assays that can reproducibly quantify the reservoir size and changes therein, in participant’s blood during a therapeutic trial. Quantification of the latent HIV viral reservoir requires a highly sensitive, cost-effective assay capable of measuring the low frequency of rCD4+ T cells carrying functional provirus. Preferably, such an assay should be such that it can be adopted for high throughput and could be adopted under conditions for use in large-scale clinical trials. While PCR-based assays are commonly used to quantify pro-viral DNA or intracellular RNA transcript, they cannot distinguish between replication-competent and defective proviruses. We have recently published a study where a reporter cell-based assay (termed TZA or TZM-bl based quantitative assay) was used to quantify inducible replication-competent latent HIV-1 in blood. This assay is more sensitive, cost-efficient, and faster than available technology, including the quantitative viral outgrowth assay or the Q-VOA. Using this assay, we show that the size of the inducible latent HIV-1 reservoir in virally suppressed participants on ART is approximately 70-fold larger than previous estimates. We describe here in detail an optimized method to quantitate latently infected cells using the TZA.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC6699756&blobtype=pdf
Volume Number	9
PubMed Central reference number	PMC6699756
Issue Number	10
PubMed reference number	31428662
Journal	Bio-protocol [Bio Protoc]
e-ISSN	23318325
DOI	10.21769/bioprotoc.3232
Language	English
Publisher	Bio-Protocol
Publisher Date	2019-05-01
Publisher Place	USA
Access Restriction	Open
Rights License	Copyright © 2019 The Authors; exclusive licensee Bio-protocol LLC.
Subject Keyword	TZM-bl cells TZA assay Latent reservoir Latent HIV-1 Quantification of latent reservoir Inducible virus Replication competent virus
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Neuroscience Plant Science Biochemistry, Genetics and Molecular Biology

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