A Novel Rapid and Highly Sensitive Cell Based System for the Detection and Characterization of HIV

Unprecedented rapidity Infection can be detected as early as 17 hours post-infection, compared to 72 hours for current methods Viral tropism can be determined in 2-4 days, compared to 3-4 weeks for current methods Extremely sensitive, with as little as 0.0625 MOI required Infection can be traced over time since the reporter protein is secreted CD4 and co-receptor densities can be precisely tuned to physiological levels Readily adaptable to high throughput formats, such as 96- and 384-well plates Very low background signal, driven by the reporter protein's dependence on both tat and rev expression by the virus

Assay for HIV entry, infection and replication efficiency over time under various laboratory conditions Calculate IC50 values for antiretroviral drugs Test for acute viral infections (not just HIV) Characterize patient-derived viruses' tropisms to guide therapeutic options This would represent the first alternative to the only product currently available Conduct basic research on antiretroviral drug mechanisms Monitor relative resistance or sensitivity to neutralizing antibodies

Dr. Benhur Lee and colleagues in the UCLA Department of Microbiology, Immunology and Molecular Genetics have developed a novel system to detect and characterize HIV with unprecedented sensitivity and rapidity. By engineering a cell line with precise control over CD4 and CCR5 expression, the researchers enable comprehensive characterization of viral entry efficiency as a function of receptor density. Combining this with a secreted tat-rev dependent reporter protein 1,000-fold brighter than luciferase, the result is GGR, a novel cell line that can detect pseudotype and replication competent HIV in less time and with higher sensitivity compared to what is currently available.

Patent Number: WO2013142341A1
Application Number: WO2013US32178A
Inventor: LEE, Benhur | CHIKERE, Kelechi | CHOU, Tom
Priority Date: 20 Mar 2012
Priority Number: WO2013142341A1
Application Date: 15 Mar 2013
Publication Date: 26 Sep 2013
IPC Current: C12N000510 | C12N001562 | C12N001563 | C12N001586 | C12Q000166
Assignee Applicant: The Regents of the University of California
Title: A NOVEL RAPID AND HIGHLY SENSITIVE CELL BASED SYSTEM FOR THE DETECTION AND CHARACTERIZATION OF HIV | NOUVEAU SYSTÈME RAPIDE ET TRÈS SENSIBLE BASÉ SUR DES CELLULES POUR LA DÉTECTION ET LA CARACTÉRISATION DU VIH
Usefulness: A NOVEL RAPID AND HIGHLY SENSITIVE CELL BASED SYSTEM FOR THE DETECTION AND CHARACTERIZATION OF HIV | NOUVEAU SYSTÈME RAPIDE ET TRÈS SENSIBLE BASÉ SUR DES CELLULES POUR LA DÉTECTION ET LA CARACTÉRISATION DU VIH
Summary: For detecting pseudotype or replication competent HIV, from cloned or uncloned isloates, in cell media or human serum via an HIV tat-rev dependent GFP-Gaussia luciferase Reporter cell line (GGR); for assaying a biological sample; for controlling the expression of any two genes of interest (all claimed); for characterizing the entry phenotype of HIV envelope genes; for detecting and examining primary HIV samples in the context of laboratory research, clinical trial monitoring, or medical diagnostics; for determining functional HIV viral load, responsiveness to HIV treatment, or characterizing viral co-receptor usage.
Novelty: New cell line comprising tat/rev dependent reporter lentiviral vector that expresses secreted Gaussia Luciferase in tandem with enhanced green fluorescent protein for detecting pseudotype or replication competent HIV

診斷/治療

人類免疫缺陷病毒

9719127

State Of Development

The GGR indicator cell line has been engineered in its entirety, and the researchers have demonstrated acute sensitivity of the cell line to infectious HIV detection with pseudotype and replication-competent viruses.   

Background

AIDS, the disease caused by the virus HIV, represents a devastating global pandemic. According to a United Nations report in 2010, HIV has killed nearly 30 million people worldwide, with over 2.5 million additional infections each year. Detecting HIV particles is critical not only to patient diagnosis, but also for basic and clinical research, the source of future therapies. Unfortunately, current methods are severely lacking. Phenotypic testing can take over a month to complete and only reports a single time point. Another system widely used for research employs cell lines that express CD4 and co-receptors at abnormally high levels, rendering results of questionable physiological relevance. Patients, physicians, and researchers alike would benefit greatly from a new method of detecting and characterizing HIV; one that is rapid, sensitive, adaptable, and most importantly, physiologically accurate.     

Related Materials

Affinofile profiling: how efficiency of CD4/CCR5 usage impacts the biological and pathogenic phenotype of HIV. Virology (2013)

Additional Technologies by these Inventors

• Optimized Matrix Based Virus-like Particle Entry and Budding Assay for Highly Pathogenic Viruses
• Preparation and Activity of Novel Photosensitizer Acting as a Broad Spectrum Antiviral Agent Against Enveloped Viruses

Tech ID/UC Case

23054/2012-524-0

Related Cases

2012-524-0

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