Anti-virals

Literature highlights

Mitochondria are of particular importance to cell health as they produce the ATP necessary for cell energy. In 2005, Seth et al. demonstrated the role of the mitochondria in cellular innate antiviral immunity and the close interconnection between the mitochondria and the antiviral immune response.

The discovery of a protein named "mitochondrial antiviral signalling protein, MAVS"1‑4 also showed the deep relation between the two. MAVS is reponsible for mitochondria localization, dimerization and for activating the intracellular signaling pathway. The three main mitochondrial pathways—β-oxidation, cycle and oxidative phosphorylation—have shown a connection with viral life cycles,5 and could reveal important information for viral therapy. Cleavage of MAVS by HCV was shown to result in the loss of MAVS mitochondrial localisation, thereby disrupting its function in interferion induction.6

Moreover, in 2015, West et al. demonstrated that "Mitochondrial DNA (mtDNA) stress primes the antiviral innate immune response." mtDNA represents a sensitive biomarker for cell adaptability.7 Furthermore, excessive generation of reactive oxygen species (ROS) activates cells' antibacterial and antiviral defense mechanisms, contributing to innate immune activation.8

All these recent discoveries show the importance and the necessity to study the effects of anti-virals on mitochondrial DNA content and function with respect to extracellular medium.

1. Kawai T., Takahashi K., Sato S., Coban C., Kumar H., Kato H., Ishii K. J., Takeuchi O., and Akira S. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nature Immunology. 2005; 6: 981–988.

2. Meylan E., Curran J., Hofmann K., Moradpour D., Binder M., Bartenschlager R., Tschopp J. Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature. 2005; 437(7062): 1167–1172.

3. Seth R. B., Sun L., Ea C. K., and Chen, Z. J. Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3. Cell. 2005; 122(5): 669–682.

4. Xu L.-G., Wang Y.-Y., Han K.-J., Li L.-Y., Zhai Z., and Shu H.-B. VISA is an adapter protein required for virus-triggered IFN-beta signaling. Molecular Cell. 2005; 19(6): 727–740.

5. Claus C., and Liebert U.G., A renewed focus on the interplay between viruses and mitochondrial metabolism. Archives of Virology. 2014; 159(6): 1261-1277.

6. Li X. D., Sun L., Seth R. B., Pineda G., and Chen Z.J. Hepatitis C virus protease NS3/4A cleaves mitochondrial antiviral signaling protein of the mitochondria to evade innate immunity. PNAS, 2005, 102: 17717-17722.

7. West A. P., Khoury-Hanold W., Staron M., Tal M. C., Pineda C. M., Lang S. M., Bestwick M., Duguay B. A., Raimundo N., MacDuff D. A., Kaech S. M., Smiley J. R., Means R. E., Iwasaki A., and Shadel G. S. Mitochondrial DNA stress primes the antiviral innate immune response. Nature. 2015; 520(7548): 553-557.

8. West A. P., Shadel D. S., and Ghosh S. Mitochondria in innate immune response. Nature reviews immunology. 2011; 11: 389-402.

All our assays are fully customizable and can be adapted to meet your specific needs. Contact us to learn how our technologies might be of value to you!

Which technologies to use

mtDNA content

We offer a fully quantitative High Content Analysis (HCA) reading of mtDNA content to help you screen for anti-viral drugs. You can also assess the impact of your drug on mtDNA content as a potential source of unwanted toxicity, as drug-induced reduction in mtDNA content is associated with severe adverse drug reactions.

This package consists of the multiplexed and integrated measurements, in a same cell, of:

  • Mitochondrial DNA content
  • Mitochondrial mass
  • Mitochondrial biogenesis
  • Cell viability
  • Nucleus integrity and morphology, an apoptotic marker (optional)
Quantitative Western blot analyses

Use this technology to:

  • Evaluate our antivirals' impact on pertinent mitochondrial targets such as MAVS.
  • Identify intracellular signal pathways activated.

BBS + package

ICDD's Bioenergetic Balance Screen (BBS), available in HTS/dose response studies, helps you evaluate your compound's impact on β-oxidation, oxidative phosphorylation (OXPHOS) and the tricarboxylic (TCA) cycle.

The BBS+ package is the multiplexed and integrated measurements, in a same cell, of:

  • Oxygen consumption
  • ATP production
  • Glycolysis level
  • TCA cycle turning
  • Cell viability

Redox status

Our Redox status package, avalaible in HTS/dose response studies, is the measurement of (any or all):

  • ROS mitochondrial production
  • Cytoplasm ROS accumulation
  • MnSOD Cu/ZnSOD activities
  • Catalase activity
  • Total GSH activity
  • Lipid peroxidation level (TBARS)
  • Protein carbonylation
  • Cell viability