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ORIGINAL RESEARCH ARTICLE

Proteomic Analysis of Endothelial Cell Autoantigens Recognized by Anti-Dengue Virus Nonstructural Protein 1 Antibodies

Hsien-Jen Cheng^*, Chiou-Feng Lin, Huan-Yao Lei, Hsiao-Sheng Liu, Trai-Ming Yeh, Yueh-Hsia Luo^* and Yee-Shin Lin^,1

^* Institute of Basic Medical Sciences, Institute of Clinical Medicine, Department of Microbiology and Immunology, and Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University Medical College, Tainan 701, Taiwan

To whom requests for reprints should be addressed at ¹ Department of Microbiology and Immunology, National Cheng Kung University Medical College, 1 University Road, Tainan 701, Taiwan. E-mail: yslin1{at}mail.ncku.edu.tw

We previously showed the occurrence of autoimmune responses in dengue virus (DV) infection, which has potential implications for the pathogenesis of dengue hemorrhagic syndrome. In the present study, we have used a proteomic analysis to identify several candidate proteins on HMEC-1 endothelial cells recognized by anti-DV nonstructural protein 1 (NS1) antibodies. The target proteins, including ATP synthase β chain, protein disulfide isomerase, vimentin, and heat shock protein 60, co-localize with anti-NS1 binding sites on nonfixed HMEC-1 cells using immunohistochemical double staining and confocal microscopy. The cross-reactivity of anti-target protein antibodies with HMEC-1 cells was inhibited by NS1 protein pre-absorption. Furthermore, a cross-reactive epitope on NS1 amino acid residues 311–330 (P311–330) was predicted using homologous sequence alignment. The reactivity of dengue hemorrhagic patient sera with HMEC-1 cells was blocked by synthetic peptide P311–330 pre-absorption. Taken together, our results identify putative targets on endothelial cells recognized by anti-DV NS1 antibodies, where NS1 P311–330 possesses the shared epitope.

Key Words: proteomic analysis • autoantibodies • endothelial cell autoantigens • dengue virus