Inactivation of Chikungunya virus by 1,5 iodonapthyl azide
© Sharma et al.; licensee BioMed Central Ltd. 2012
Received: 24 May 2012
Accepted: 15 November 2012
Published: 4 December 2012
Chikungunya virus (CHIKV) is an arthropod borne alphavirus of the family Togaviridae. CHIKV is a reemerging virus for which there is no safe prophylactic vaccine. A live attenuated strain of CHIKV, CHIK181/25, was previously demonstrated to be highly immunogenic in humans, however, it showed residual virulence causing transient arthralgia.
In this study, we demonstrate the complete inactivation of CHIKV181/25 by 1,5 iodonapthyl azide (INA). No cytopathic effect and virus replication was observed in cells infected with the INA-inactivated CHIKV. However, a reduction in the INA-inactivated CHIK virus-antibody binding capacity was observed by western blot analysis.
INA completely inactivated CHIKV and can further be explored for developing an inactivated-CHIKV vaccine.
KeywordsChikungunya virus Inactivated vaccine Iodonapthyl azide
Earlier we have shown that 1,5 iodonapthyl azide (INA) and UV irradiation completely inactivated the Venezuelan equine encephalitis virus (VEEV)[1, 2]. In this study, we have extended that approach to evaluate the inactivation of Chikungunya virus (CHIKV). CHIKV is an arthropod borne alphavirus of the family Togaviridae. It is a reemerging virus that has caused wide spread outbreak in the nations surrounding the Indian Ocean and Africa since 2004. Among the alphaviruses, CHIKV is one of the most important human pathogens due to the frequent outbreaks worldwide, causing large scale morbidity and economic losses. CHIKV infection has been reported in the travelers returning from the epidemic regions, thereby indicating a potential of dissemination to the previously known naïve regions for CHIKV. Further, E1-A226V mutation in the CHIKV strain of 2005–2006 outbreak has increased the susceptibility of Aedes albopictus, which is endemic to the previously CHIKV naïve regions[5, 6]. Therefore, there is an ever increasing risk of CHIKV outbreak in the areas such as the United States of America, where CHIKV is not endemic. Such potential outbreak may be devastating due to lack of immunity to CHIKV in the endemic population. There are no specific drugs against CHIKV and patients are symptomatically treated with non steroid anti-inflammatory drugs. In the absence of antiviral drugs, preventive measures that include effective CHIKV vaccine are urgently needed. The United States Army had developed and tested a live attenuated strain of CHIKV, CHIKV181/25 for vaccine application. CHIKV181/25 demonstrated an excellent immunogenic profile (98% seroconversion), however, transient arthralgia was observed in about 8% of vaccine recipients. Several other approaches such as chimeric CHIKV on other alphavirus backbone, CHIK virus like particles, formalin inactivated CHIKV, DNA immunization and passive immunization with human polyvalent immunoglobulin have also been tested[9–13].
INA is a photoactive hydrophobic azide molecule that sequesters in the hydrophobic domain of the biomembranes and binds to the membrane proteins upon irradiation with long wavelength ultra-violet (UV) light. It was initially used for labeling the biological membranes. Interaction of INA with proteins inactivates the membrane protein while conserving the ectodomain protruding outside the membrane. This property of INA has been used to inactivate influenza virus, Ebola virus, VEEV, vaccinia virus, pixuna virus, simian and human immunodeficiency viruses and provide potential vaccine candidates[1, 2, 15–18]. In this study, we have evaluated the dose dependent inactivation of CHIKV181/25 by INA.
Our findings are important as CHIKV causes an ongoing outbreak affecting millions of people around the globe. An effective CHIKV vaccine is urgently needed to control the spread of CHIKV. Live attenuated CHIKV181/25 was shown to have an excellent immunogenicity; however, the clinical trial was halted due to issues with residual virulence causing transient arthralgia in the vaccine recipients. The INA-inactivated CHIKV181/25 formulation may address the issue of residual virulence that is associated with live attenuated CHIKV181/25. INA-inactivation resulted in a relatively less binding capacity of CHIKV181/25 to the neutralizing polyclonal anti-CHIKV E2 glycoprotein. This raises the question whether there may be some loss of antigenicity upon inactivation with INA. However, INA-inactivated CHIKV may still elicit a protective response against virulent CHIKV infection as shown with other INA-inactivated enveloped viruses such as VEEV, Ebola and Influenza virus. Chemical inactivation of CHIKV for vaccine purposes has been previously reported[11, 19]. Formalin inactivated CHIKV was shown to induce neutralizing antibody and reduce homologous virulent virus titer. Further testing of INA-inactivated CHIKV in the animal model will be needed for testing the protective efficacy of INA-inactivated CHIKV181/25 as a vaccine candidate, and its advantage over current approaches of generating inactivated vaccines. To our knowledge this is the first study to show a complete inactivation of CHIKV181/25 using INA.
1, 5 iodonapthyl azide
- BHK cells:
Baby hamster kidney cells
Phosphate buffered saline
Bovine serum albumin
Reverse Transcription-Polymerase chain reaction.
These studies were supported by JSTO-CBD/DTRA Contract/Grant/Intergovernmental Project Order/Project # 4.10019_07_US_B and USAMRIID contract No. G174QH. Authors acknowledge the technical help from Mr. Gautham Sukumaran and Mr. Raghavendar Chandran. INA was provided by Dr. Robert Blumenthal, CCR Nanobiology Program, Centre for Cancer Research, National Cancer Research Institute, Frederick, MD under MTA. The opinions or assertions contained herein are the scientific views of the authors and should not be construed as official or necessarily reflecting the views or policies of the Uniformed Services University of the Health Sciences or the Department of Defense, USA.
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