Identification of Disease-Specific B-Cell Epitopes for Antibodies from Dengue-Infected Human Serum Samples
Date Issued
2005
Date
2005
Author(s)
Li, Pi-Chun
DOI
en-US
Abstract
Dengue is an expanding public health problem in the tropics and subtropical area and it’s the most important arboviral disease of humans. An estimated 50-100 million dengue infections and 500,000 DHF cases occur annually (World Health Organisation, 2000), and its effective vaccine still remains elusive.
Recognition of viral protein epitopes is required for the development of useful therapies and understanding the pathogenesis of the disease. To investigate disease-specific B-cell epitopes, human serum antibodies from DEN-2 and DEN-4 infected patients were biopanning with phage-displayed random peptide libraries. Immunopositive phage clones were proved to bind specifically to serum samples of dengue patients but not to normal human serum samples. The binding reactivity of the selected phage clones was divided into three different groups. Some epitopes had higher reactivity to DHF serum samples and the other had greater preference to DF. The third group of the selected phage clones recognized both DHF and DF serum samples. These phage-borne peptides had consensus motifs, which corresponded to amino acid sequences of dengue virus, included capsid (C), envelope (E) and nonstructural (NS) proteins. One of these phage clones, DP1-1-54, had higher reactivity to DHF serum samples which corresponded to amino acid sequences of dengue virus E protein; DP3-25 had greater preference to DF; DP1-2-10, DP2-2-7, recognized both DHF and DF. DP1-2-10 and DP3-25 had no corresponded to amino acid sequences reported for dengue virus. DP2-2-7 had corresponded to C protein.
To apply phage-displayed epitopes for diagnosis of dengue patients, a recombinant GST-epitope-fusion protein was produced. Immunopositive phage clones and recombinant GST-epitope-fusion protein demonstrated clinical diagnostic potential by reacting with serum from patients with dengue. Our epitope-based serologic test may be useful in laboratory detection of dengue infection. Furthermore, identification of dengue specific epitopes is important in studying the virus-antibody interactions at a molecular level and address the role of antibodies in the pathogenesis of DHF.
Subjects
登革
B細胞抗原決定位
登革出血熱
套膜蛋白
dengue
B cell epitope
DHF
E protein
SDGs
Type
other
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