Abstract
BackgroundHuman adenoviruses (HAdVs) have been implicated as important agents in a wide range of human illnesses. To date, 58 distinct HAdV serotypes have been identified and can be grouped into six species. For the immunological diagnosis of adenoviruses, the hexon protein, a structural protein, has been used. The potential of other HAdV proteins has not been fully addressed.Methodology/Principal FindingsIn this study, a nonstructural antigenic protein, the DNA binding protein (DBP) of human adenovirus 5 and 35 (Ad5, Ad35) - was identified using immunoproteomic technology. The expression of Ad5 and Ad35 DBP in insect cells could be detected by rhesus monkey serum antibodies and healthy adult human serum positive for Ad5 and Ad35. Recombinant DBPs elicited high titer antibodies in mice. Their conserved domain displayed immunological cross-reactions with heterologous DBP antibodies in Western blot assays. DBP-IgM ELISA showed higher sensitivity adenovirus IgM detection than the commercial Adenovirus IgM Human ELISA Kit. A Western blot method developed based on Ad5 DBP was highly consistent with (χ2 = 44.9, P<0.01) the Western blot assay for the hexon protein in the detection of IgG, but proved even more sensitive.Conclusions/SignificanceThe HAdV nonstructural protein DBP is an antigenic protein that could serve as an alternative common antigen for adenovirus diagnosis.
Highlights
Viral structural proteins, which generally elicit strong host immune responses, have been widely used in vaccine and diagnostic reagent development
The differentially expressed Human adenoviruses (HAdVs) proteins identified by 2DEPAGE and Western blot were further analyzed using MALDITOF/TOF
As the nonstructural protein DNA binding protein (DBP) was identified in both Ad5 and Ad35 infected HEK293 cells, our findings suggest that this nonstructural protein may function as an additional HAdV antigen
Summary
Viral structural proteins, which generally elicit strong host immune responses, have been widely used in vaccine and diagnostic reagent development. Viral nonstructural proteins can stimulate unique immune responses during certain infectious processes [4]. This antigenicity and immunogenicity of viral nonstructural proteins aids vaccine design and clinical diagnosis of viral infections [4,5]. At least 58 distinct serotypes have been identified [8] These serotypes have been grouped into six species HAdV-A to HAdV-F [8] based on their immunological, biological, and biochemical characteristics. 58 distinct HAdV serotypes have been identified and can be grouped into six species. The potential of other HAdV proteins has not been fully addressed
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