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Abstract
The non-structural protein-1 (NS1) of dengue virus (DENV) contributes to several functions related to dengue disease pathogenesis as well as diagnostic applications. Antibodies against DENV NS1 can cross-react with other co-circulating flaviviruses, which may lead to incorrect diagnosis. Herein, five anti-DENV NS1 monoclonal antibodies (mAbs) were investigated. Four of them (1F11, 2E3, 1B2, and 4D2) cross-react with NS1 of all four DENV serotypes (pan-DENV mAbs), whereas the other (2E11) also reacts with NS1 of other flaviviruses (flavi-cross-reactive mAb). The binding epitopes recognized by these mAbs were found to overlap a region located on the disordered loop of the NS1 wing domain (amino acid residues 104 to 123). Fine epitope mapping employing phage display technology and alanine-substituted DENV2 NS1 mutants indicates the critical binding residues W115, K116, and K120 for the 2E11 mAb, which are conserved among flaviviruses. In contrast, the critical binding residues of four pan-DENV mAbs include both flavi-conserved residues (W115 to G119) and DENV-conserved flanking residues (K112, Y113, S114 and A121, K122). Our results highlight DENV-conserved residues in cross-reactive epitopes that distinguish pan-DENV antibodies from the flavi-cross-reactive antibody. These antibodies can be potentially applied to differential diagnosis of DENV from other flavivirus infections.
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1 National Science and Technology Development Agency (NSTDA), Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathum Thani, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408); BIOTEC, NSTDA, Medical Biotechnology Research Unit, Bangkok, Thailand (GRID:grid.419250.b)
2 Mahidol University, Graduate Program, Department of Immunology, Faculty of Medicine, Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
3 National Science and Technology Development Agency (NSTDA), Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathum Thani, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408); BIOTEC, NSTDA, Medical Biotechnology Research Unit, Bangkok, Thailand (GRID:grid.419250.b); Mahidol University, Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
4 BIOTEC, NSTDA, Medical Biotechnology Research Unit, Bangkok, Thailand (GRID:grid.419250.b); Mahidol University, Division of Dengue Hemorrhagic Fever Research, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490); Mahidol University, Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
5 Mahidol University, Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
6 BIOTEC, NSTDA, Biomedical Technology Research Center, Chiang Mai, Thailand (GRID:grid.10223.32); Chiang Mai University, Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai, Thailand (GRID:grid.7132.7) (ISNI:0000 0000 9039 7662)