Dengue: Fastest Emerging Infection

by Orville Yu


WHO declares that dengue is one of the fastest-emerging infections    in the world, [but research focus and funding have decreased].

DENGUE fever and dengue hemorrhagic fever (DHF) are diseases caused by a family of viruses that are transmitted by mosquitoes. Dengue viruses infect nearly 100 million people each year in more than 100 countries. Seventy percent of these come from the Asia-Pacific. WHO said "much of Asia continues to be in the grip of dengue fever, with the number of hospitalizations and severe cases growing." Some countries have reported significant increases in dengue outbreaks as compared to the same period last year. Among the badly affected countries cited are the Philippines, Lao People's Democratic Republic, Malaysia, and Vietnam.

Dengue hemorrhagic fever (DHF) is a more severe form of the viral illness. Manifestations include headache, fever, rash, and evidence of hemorrhage in the body. Small red or purple blisters under the skin, bleeding in the nose or gums are all possible signs of hemorrhage. This form of dengue fever can be life-threatening and can progress to the most severe form of the illness, dengue shock syndrome (DSS). The mortality (death) rate with DHF is significant. It ranges from 6%-30%. Most deaths occur in children.

The dengue virus (DENV)
The disease is caused by four closely related virus serotypes of the genus Flavivirus, family Flaviviridae. Each serotype is sufficiently different that there is no cross-protection and epidemics caused by multiple serotypes can occur. Dengue is transmitted to humans by the Aedes aegypti mosquito. Because it is caused by one of four serotypes of virus, it is possible to get dengue fever multiple times. However, an attack of dengue produces immunity for a lifetime to that particular serotype to which the patient was exposed.



How dengue virus invades a cell

Dengue is an enveloped virus, which means it's surrounded by a protein layer and a lipid bilayer. Inside this viral envelope is a protein coat called a capsid, and the RNA genome. To infect a host cell, the dengue virus must find a way to penetrate the cell's plasma membrane. 


Dengue virus enters a host cell when the viral envelope glycoprotein, E, binds to a receptor. Its major envelope glycoprotein, E, mediates viral attachment and entry by membrane fusion. A crystal structure of the soluble domain of E from dengue virus type 2 reveals a hydrophobic pocket lined by residues that influence the pH threshold for fusion. The pocket, which accepts a hydrophobic ligand, opens and closes through a conformational shift in a beta-hairpin at the interface between two domains. Once inside, it must find a way to fuse yet another cellular membrane—the host cell's endosomal membrane—with its own so that the virus' RNA cargo can be released.


This visualization adapted from the University of Massachusetts Medical School represents the process by which a dengue virus releases its genetic contents inside a host cell, allowing viral replication. The key players are proteins found on the viral surface called envelope proteins, which change their structure when the virus is endocytosed, or taken inside the cell. This structural change enables the viral membrane to fuse with the endosomal membrane, and the virus' RNA to enter the cytoplasm of a host cell. There, ribosomes will make viral proteins that will spread to other cells through the replication and secretion of new viral particles.

Treatment for dengue fever

Because dengue fever is caused by a virus, there is no specific medicine or antibiotic to treat it. There are no available vaccines or antivirals against DENV. Currently, vector control is the only method for prevention of the disease. Development of a successful vaccine would require for it to be effective against all four DENV serotypes, economical, and provide long-term protection. For typical dengue, the treatment is purely concerned with relief of the symptoms (symptomatic). Rest and fluid intake for adequate hydration is important. Aspirin and nonsteroidal anti-inflammatory drugs should only be taken under a doctor's supervision because of the possibility of worsening hemorrhagic complications. Paracetamol, acetaminophen and codeine may be given for severe headache and for the joint and muscle pain.

Dengue cases are increasing
The WHO pointed out a number of factors that have caused the increase in dengue cases around the world. These include higher temperatures and rainfall, growing population densities, and greater international travel. So far, there is no firm evidence that points out global warming as the ultimate culprit, climate change has played an important role. Because of climate change, mosquitoes are now found in areas where they were once uncommon. Dengue may also be transmitted via infected blood products and in countries such as Singapore, where dengue is endemic, the risk was estimated to be between 1.6 and 6 per 10,000 blood transfusions.

Of the countries in the table, the only nations with lower cases and deaths so far in 2010 are Singapore, Mexico and the United States. In many undeveloped regions, including parts of India, Philippines and Malaysia, authorities do not have adequate facilities to detect dengue cases. 



Current research efforts on dengue
A health problem of this scope should be regarded as high priority and should have attracted ample funding from donors and national authorities. But such is not the case. Historical review reveals that there is a greater number of laboratories and a greater allocation of resources to dengue research 30-50 years ago than there is today.

Molecular replication mechanism of the DENV.
In 2006 a group of Argentine scientists discovered the molecular replication mechanism of the virus, which could be specifically attacked by disrupting the viral RNA polymerase.

“At first we were puzzled by the cyclization feature of this virus. We now recognize that it serves a role in bringing the promoter near the initiation site”, a statement from Andrea V. Gamarnik, a virologist from Leloir Institute Foundation in Buenos Aires. They have described how a viral enzyme recognizes and amplifies the genetic material needed to assemble new dengue viruses. Their findings provide the first model for RNA replication in the family of viruses that includes West Nile, St. Louis encephalitis, and hepatitis C.

Molecules as inhibitors to stop replication.
Other studies are working on the search of possible small molecules that can bind at the ligand-binding pocket present in the dengue virus. The binding of such molecule can act as inhibitors to stop infection.


Further research done by this group proposed a fusion mechanism driven by essentially irreversible conformational changes in the viral envelope glycoprotein, E, and facilitated by fusion-loop insertion into the outer bilayer leaflet. Specific features of the folded-back structure suggest strategies for inhibiting flavivirus entry.



Another study involving dengue drug discovery is trying to develop drugs that inhibit the viral NS3 protease. The NS3 protease is an enzyme critical for virus replication, and its amino acid sequence and atomic structure are very similar among the different disease–causing flaviviruses. Since the atomic structure of the NS3 protease is known, we can utilize advanced structure–based computational drug discovery methods to identify small molecule protease inhibitors. Dr. Stan Watowich and his research team at The University of Texas Medical Branch (Galveston, Texas, USA) have made significant progress in this direction, having discovered compounds that inhibit dengue and West Nile virus proteases and prevent virus replication in cell culture. However, additional drug candidates need to be discovered to improve the likelihood of converting drug leads into approved drugs for treating flavivirus infections.

Recently, the company, NanoViricides, Inc. developed a library of chemical ligands that are expected to bind to the dengue virus envelope proteins of several different subtypes of dengue viruses. These ligands were developed using the results of sophisticated, well established, molecular modeling software. A number of candidate nanoviricides that are capable of attacking the dengue virus were created using these ligands. A "nanoviricide" is a chemical substance made by covalently attaching a number of copies of a virus-binding ligand to a base polymeric micelle, that the Company calls TheraCour®. It is believed that when a nanoviricide binds to a virus particle, the interaction would extend to the binding of a large number of ligands to the virus surface, and the flexible nanomicelle would then engulf the virus, rendering it incapable of infecting a cell.


Cited Literature

Modis, Y., Ogata, S., Clements, D. & Harrison, S.C. Structure of the dengue virus envelope glycoprotein after membrane fusion. Nature, 427: 313-319 (2004).

Kuhn RJ, Zhang W, Rossmann MG, et al. Cell 2002, 108:717-725.

Modis, Y., Ogata, S., Clements, D. & Harrison, S.C. A ligand-binding pocket in the dengue virus envelope glycoprotein. PNAS, 100, 12: 6986-6991 (2003).

World Health Organization (on dengue). Can be accessed at <www.who.int> by searching dengue or dengue virus. Last accessed: October 6, 2010.

WHO Dengue and dengue hemorrhagic fever Fact Sheet No. 117, March 2009

Medicinenet (on dengue). Can be accessed at <www.medicinenet.com> by searching dengue or dengue virus. Last accessed: October 7, 2010.

The Medical News <news-medical.net>. Anti-dengue drug candidates demonstrate efficacy in preliminary cell culture studies.  June 2010.