ViroVision™ HIV Reporter Systems

ViroVision™ Reporter Systems represent a significant advacement for HIV basic research. The ViroVision™ Reporter System is composed of two components: ViroVision™ Reporter Cells and ViroVision™ Infection Medium (patent pending).
 

ViroVision™ Reporter Cells: ViroVision™ Reporter Cells are T-cell derived cell lines, engineeered with the HIV-specific Rev Response Element (RRE).  They contain a GFP, Luc, or GFP/Luc reporter. The RRE eliminates unwanted background signal incurred with LTR-only HIV Indicator cells, enabling a broad range of applications.
 

ViroVision™ Infection Medium: ViroVision™ Infection Medium (patent pending) is a unique, HIV-specific infection enhancer that does not contain DEAE-dextran. ViroVision™ Infection Medium works with any HIV-receptive cell line and will increase HIV infectivity rates 3 - 10 fold over native infection rates.


ViroVision™ HIV Reporter Systems

A new generation of HIV Indicator Cells:

ViroVision™ HIV Reporter Cells represent a significant advancement for HIV basic research. Engineered with the HIV-specific Rev Response Element (RRE), ViroVision™ HRC provide unparalleled sensitivity & specificity for a broad range of applications including screening broadly neutralizing antibodies and anti-HIV drugs. Derived from CD4 T-cells, ViroVision™ HRC express more native levels of HIV receptors and are more natural HIV targets with broad susceptibility to X4, R5, and primary HIV isolates. With GFP, Luc, or GFP/Luc detection options, ViroVision™ HRC provide a versatile and flexible platform for your HIV studies.

For a complete selection guide, click here.

ViroVison™ Brochure: ViroVison™ Protocol: Limited Use License:

ViroVision™ HRC Applications

ViroVision™ cells will allow you to more easily perform:
  • TCID50 assays: Routine HIV infectivity & quantification
  • Anti-HIV drug screenings via one-step infection
  • Routine EC50/LD50 quantifications of anti-HIV compounds
  • Screenings for broadly neutralizing antibody (bnAB) (from laboratory and clinical research samples)
  • Quantification of neutralizing antibodies
  • HIV cell-cell transmission and HIV drug-resistance studies
  • Low-level HIV gene expression assessments
  • HIV pre-integration transcription studies
  • HIV latency and reactivation studies
  • Studies of HIV outgrowth due to reactivation
  • Determination of HIV tropisms
  • HIV+ exosome studies
ViroVision HIC system provides the ability to perform more HIV sensing applicatins.

Application:

Quantification of HIV Neutralizing Antibodies
ViroVision HIC system provides the ability to perform more HIV sensing applications.

Prior to infection, HIV(AD8), an R5 virus, was incubated with or w/o the HIV neutralizing antibody B12 (10 µg/ml final). After 1 h, Rev-A3R5-GFP cells were infected with Ab-neutralized and non-neutralized virus. Cells were washed and cultured for 48 hours. GFP expression was quantified by flow cytometer. PI = propidium iodide.

 

 

HIV Sensitivity and Specificity -- No background signal

Nearly all HICs use the LTR promoter to drive reporter expression. While responsive to HIV Tat, LTR promoters generate false positive reporter expression due to HIV-independent factors resulting in lower HIV specificity and a lower HIV detection range. Changes in cell culture conditions, the presence of mitogens, cytokines, cellular activators, and chromatin modulator may all produce high background signal in LTR reporter systems. ViroVision™ cells overcome the drawbacks of LTR reporter lines by being engineered to leverage the HIV Rev-Response Element (RRE) to regulate reporter expression. Rev is present only in HIV+ cells. The high stringency that Rev imposes on the reporter dramatically decreases background signal and significantly increases sensitivity. Thus, ViroVision™ cells can accurately detect low-levels of HIV activity, even in the presence of environmental factors that generate signal in cells containing a reporter driven by an LTR-only promoter.

The Rev-reporter system employs both the LTR and Rev response element from HIV. ViroVision™ Cells carry stably integrated reporter constructs that are derived from the HIV genome. The incorporation of the RRE and multiple authentic HIV splicing sites permits reporter expression only from the non-spliced and singly-spliced transcripts in the presence of Rev.
Rev-reporters eliminate noise incurred with standard LTR reporter systems. ViroVision™ Cells are more HIV-specific compared with LTR-GFP reporter cells. Without HIV, ViroVision™ Cells have undetectable GFP, whereas LTR-GFP cells have high background GFP. ViroVision™ Cells do not respond to PMA stimulation (100 ng/ml). For LTR-GFP cells, PMA induces high levels of GFP signal.

Tired of Being Blue?

Go Green or Catch a Firefly Instead...

ViroVision HIC system provides the ability to perform more HIV sensing applicatins.

Have you ever mis-timed your assays only to have all your cells turn blue? Does the picture to the right look all to familiar? ViroVision™ Cells eliminate the potential to waste time and money because your current indicator cells turned blue without HIV infection or because you did not perfectly time your X-Gal staining. With unparalleled specificity & sensitivity, ViroVision™ Cells come with a GFP, Luciferase or a combination GFP/Luciferase reporters that provide you with ease of use and flexibility.

 

ViroVision HIC system provides the ability to perform more HIV sensing applicatins. ViroVision HIC system provides the ability to perform more HIV sensing applicatins. ViroVision HIC system provides the ability to perform more HIV sensing applicatins.

 

Versatile & Easy-to-Use: One Infection, 3 Readouts

Simplify and increase the efficiency of your screening process. With the ViroVision™ system, 3 readouts may be obtained with one infection. Easily screen for positive candidates based on the luciferase signal and obtain population dynamics through flow cytometry based on the fluorescent signal of GFP and/or a vital dye.

Dual-reporter Virovision HIV reporter cells allow more efficient anti-HIV drug screening.

Thoroughly Characterized

Each ViroVision™ cell has been thoroughly characterized and validated with regards to responsiveness to HIV, and sensitivity for quantifying anti-HIV inhibitors, HIV activators, and HIV neutralizing antibodies. In particular, ViroVision™ HICs are derived from human T-cells and carry physiological or near-physiological levels HIV receptors and relevant T-cell receptors, they are especially suited for quantifying HIV isolates and bnAB using laboratory and clinical research samples.

Greater Physiological Relevance

ViroVision HIV Reporter celss are more physiological representative of natural HIV targets.

ViroVision™ Cells are derived from CD4 T-cells [CEM-SS (Wu et al, 2007) & A3R5, (McLinden et al. 2013)] and further engineered with a dual LTR and Rev-dependent reporter system. Because they are derived from CD4 T-cell and do not contain a super-abundance of HIV receptors, the ViroVision™ cells provide a more physiological relevant and nAB sensitive indicator system.

 

 

Receptor cell density of A3R5 cells. ViroVision™ Cell Lines (derived from CEM-SS & A3R5 cells) do not have a super-abundance of receptors and more closely mimic natural T-cell HIV receptor densities. Source: PLOS ONE, McLinden et al. 2013, 8: 11, e7756.


The Making of ViroVision™ Cells

The HIV Rev-dependent Reporter cell lines were originally developed by Wu & Marsh at the National Institute of Health. An early version of the Rev-dependent cell, Rev-CEM, has been extensively used in multiple laboratories for studying HIV infection, anti-HIV drugs, and HIV cell-cell transmission. Multiple Rev-dependent reporter cells have been developed recently in Wu's lab at George Mason University to meet the needs of the HIV/AIDS research community.

References

  1. Wu Y, Beddall MH, Marsh JW. Rev-dependent indicator T cell line. Current HIV Research. 2007;5:395-403.
  2. Sigal A, Kim JT, Balazs AB, Dekel E, Mayo A, Milo R, et al. Cell-to-cell spread of HIV permits ongoing replication despite antiretroviral therapy. Nature. 2011;477(7362):95-8. PubMed PMID: 21849975.
  3. Yoder A, Yu D, Dong L, Iyer SR, Xu X, Kelly J, et al. HIV envelope-CXCR4 signaling activates cofilin to overcome cortical actin restriction in resting CD4 T cells. Cell. 2008;134(5):782-92. PubMed PMID: 18775311.
  4. Yu D, Wang W, Yoder A, Spear M, Wu Y. The HIV envelope but not VSV glycoprotein is capable of mediating HIV latent infection of resting CD4 T cells. PLoS Pathog. 2009;5(10):e1000633. PubMed PMID: 19851458.
  5. Spear M, Guo J, Turner A, Yu D, Wang W, Meltzer B, et al. HIV-1 triggers WAVE2 phosphorylation in primary CD4 T cells and macrophages, mediating Arp2/3-dependent nuclear migration. J Biol Chem. 2014;289(10):6949-59. PubMed PMID: 24415754; PubMed Central PMCID: PMC3945356.
  6. Yoder A, Guo J, Yu D, Cui Z, Zhang XE, Wu Y. Effects of Microtubule Modulators on HIV-1 Infection of Transformed and Resting CD4 T Cells. J Virol. 2011;85(6):3020-4. PubMed PMID: 21209111.
  7. Sloan RD, Kuhl BD, Donahue DA, Roland A, Bar-Magen T, Wainberg MA. Transcription of preintegrated HIV-1 cDNA modulates cell surface expression of major histocompatibility complex class I via Nef. J Virol. 2011;85(6):2828-36. PubMed PMID: 21209113.
  8. Sloan RD, Donahue DA, Kuhl BD, Bar-Magen T, Wainberg MA. Expression of Nef from unintegrated HIV-1 DNA downregulates cell surface CXCR4 and CCR5 on T-lymphocytes. Retrovirology. 2011;7:44. PubMed PMID: 20465832.
  9. Shuck-Lee D, Chang H, Sloan EA, Hammarskjold ML, Rekosh D. Single-nucleotide changes in the HIV Rev-response element mediate resistance to compounds that inhibit Rev function. J Virol. 2011;85(8):3940-9. PubMed PMID: 21289114.
  10. Guo J, Wang W, Yu D, Wu Y. Spinoculation triggers dynamic actin and cofilin activity facilitating HIV-1 infection of transformed and resting CD4 T cells. J Virol. 2011;85(19):9824-33. PubMed PMID: 21795326.
  11. Iyer SR, Yu D, Biancotto A, Margolis LB, Wu Y. Measurement of human immunodeficiency virus type 1 preintegration transcription by using Rev-dependent Rev-CEM cells reveals a sizable transcribing DNA population comparable to that from proviral templates. J Virol. 2009;83(17):8662-73. PubMed PMID: 19553325.

 

** ViroVison™ HIV Reporter Cells may only be obtained with a Limited Use License and is intended for Research Use Only and is not for diagnostic or therapeutic purposes. **

**Under development**

Catalog No: CUBRC0011
Description: Infection of Rev-A3R5-GFP by HIV results in GFP expression in 2-5 days. Rev-A3R5-GFP is permissive t..
Size: One vial of 5 x 10⁶ cells.
Catalog No: CUBRC0012
Description: Infection of Rev-A3R5-GFP/Luc by HIV results in GFP/Luc expression in 2-5 days. Rev-A3R5-GFP/Luc is ..
Size: One vial of 5 x 10⁶ cells.
Catalog No: CUBRC0022
Description: Infection of Rev-A3-GFP/Luc by HIV results in GFP/Luc expression in 2-5 days. Rev-A3-GFP/Luc is perm..
Size: One vial of 5 x 10⁶ cells.
Catalog No: CUBRC0031
Description: Infection of Rev-CEM-GFP by HIV results in GFP expression in 2-5 days. Rev-CEM-GFP is permissive to ..
Size: One vial of 5 x 10⁶ cells.
Catalog No: CUBRC0032
Description: Infection of Rev-CEM-GFP/Luc by HIV results in GFP expression in 2-5 days. Rev-CEM-GFP/Luc is permis..
Size: One vial of 5 x 10⁶ cells.
Catalog No: CUBRC0033
Description: Infection of Rev-CEM-Luc by HIV results in GFP expression in 2-5 days. Rev-CEM-Luc is permissive to ..
Size: One vial of 5 x 10⁶ cells.
Catalog No: CUBME0012
Description: ViroVision™ Infection Medium (patent pending) is a unique, HIV-specific infection enhancer that does..
Applications: Enhancing lenti- or retroviral transduction of target cells; Enhancing infection rates of other enveloped viruses; Facilitating recovery of infectious viruses from cell or tissue cultures; Facilitating anti-viral drug screening efficiency.
Catalog No: CUBME0011
Description: ViroVision™ Infection Medium (patent pending) is a unique, HIV-specific infection enhancer that does..
Applications: Enhancing lenti- or retroviral transduction of target cells; Enhancing infection rates of other enveloped viruses; Facilitating recovery of infectious viruses from cell or tissue cultures; Facilitating anti-viral drug screening efficiency.
Catalog No: CUBME0013
Description: ViroVision™ Infection Medium (patent pending) is a unique, HIV-specific infection enhancer that does..
Applications: Enhancing lenti- or retroviral transduction of target cells; Enhancing infection rates of other enveloped viruses; Facilitating recovery of infectious viruses from cell or tissue cultures; Facilitating anti-viral drug screening efficiency.
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