Abstract
Studies investigating West Nile virus (WNV) NS4B protein function are hindered by the lack of an antibody recognizing WNV NS4B protein. Few laboratories have produced WNV NS4B antibodies, and none have been shown to work consistently. In this report, we describe a NS4B antibody against Japanese encephalitis virus (JEV) NS4B protein that cross-reacts with the NS4B protein of WNV but not of dengue virus (DENV). This JEV NS4B antibody not only recognizes WNV NS4B in infected cells, but also recognizes the NS4B protein expressed using transfection. It is evident from this data that the JEV NS4B antibody is specific to NS4B of WNV but not to NS4B of the four DENV serotypes. The specificity of this antibody may be due to the notable differences that exist between the amino acid sequence identity and antigenic relationships within the NS4B protein of the WNV, DENV, and JEV.
Highlights
The West Nile virus (WNV) genome consists of a single-stranded, positive-sense RNA of approximately 11-kb that encodes a single polyprotein precursor, which is processed by cellular and viral-encoded proteases into three structural proteins [capsid (C), pre-membrane, envelope (E)]and seven nonstructural (NS) proteins [NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5]
The cross-reactivity of the anti-Japanese encephalitis virus (JEV) NS4B antibody procured from GeneTex, Inc. was tested by western blot (WB) analysis of total protein lysates from Vero cells infected with the Nakayama strain of JEV, the NY99 strain of WNV, and the NGC strain of dengue virus (DENV)-2
The 27-kDa protein detected in WNV infected HEK 293 cell lysates, was completely undetectable in uninfected control cell lysates at 24 hr after-infection using the anti-JEV NS4B antibody (Supplementary Figure S1B), indicating that the 27-kDa band is the
Summary
The West Nile virus (WNV) genome consists of a single-stranded, positive-sense RNA of approximately 11-kb that encodes a single polyprotein precursor, which is processed by cellular and viral-encoded proteases into three structural proteins [capsid (C), pre-membrane (prM), envelope (E)]and seven nonstructural (NS) proteins [NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5]. WNV, Japanese encephalitis virus (JEV) and the four serotypes of dengue virus (DENV) all belong to the genus. The four DENV serotypes belong in the DENV serogroup while. Stratford, Usutu, St. Louis encephalitis) of the JEV serogroup [2]. The serogroup is based on the recognition of the E proteins of these viruses. Cross-reactivity of species within or between serogroups in serological tests is a common feature of flaviviruses. These cross-reactive responses make the interpretation of serological tests such as, enzyme-linked immunosorbent assay (ELISA) and neutralization tests, difficult [3]. Antibodies against the E protein of JEV could cross-react with members of JEV serogroup as well as with some viruses
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