ABSTRACT
OBJECTIVE@#To evaluate the recognition of NS4B mimotope, as multiple antigen peptide (MAP), by dengue antibodies presents in serum samples from patients with different serotype infections.@*METHODS@#A MAP containing mimotope sequence was synthesized and used to evaluate the recognition of NS4B mimotope as MAP by a panel of 66 human sera from dengue cases by an indirect ELISA assay.@*RESULTS@#The MAP differentiated between sera from dengue viruses infected patients and sera from healthy individuals and the best reactivity was shown by serum from dengue type 3 virus patients. The recognition was more intense with serum from patients with secondary infection.@*CONCLUSIONS@#The findings suggest the potential use of NS4B mimotope on the development of a multi-epitope diagnostic tool. These results are important for further immunogenicity studies.
ABSTRACT
Objective: To evaluate the recognition of NS4B mimotope, as multiple antigen peptide (MAP), by dengue antibodies presents in serum samples from patients with different serotype infections. Methods: A MAP containing mimotope sequence was synthesized and used to evaluate the recognition of NS4B mimotope as MAP by a panel of 66 human sera from dengue cases by an indirect ELISA assay. Results: The MAP differentiated between sera from dengue viruses infected patients and sera from healthy individuals and the best reactivity was shown by serum from dengue type 3 virus patients. The recognition was more intense with serum from patients with secondary infection. Conclusions: The findings suggest the potential use of NS4B mimotope on the development of a multi-epitope diagnostic tool. These results are important for further immunogenicity studies.
ABSTRACT
Background: Dengue is the most important human viral disease transmitted by arthropod vectors. The availability of random peptide libraries (RPL) displayed on phage has provided a powerful tool for selecting sequences that mimic epitopes from microorganisms that are useful for diagnostic and vaccine development purposes. In this paper, we describe peptides that resemble the antigenic structure of B-cell epitopes of dengue virus identified from a phage-peptide library using human sera containing polyclonal antibodies against dengue virus. Materials and Methods: Eighteen phage clones were isolated from the phage-display peptide library, J404, by affinity selection using human antisera against dengue virus type 3. These clones were tested for reactivity by ELISA with a panel of hyperimmune ascitic fluids (HAFs) containing antibodies either against all four dengue serotypes, West Nile virus (WNV) or Eastern equine encephalitis virus (EEEV) with control ascitic fluid (NAF) used as a negative control. Results: Eight clones were recognized by HAFs against the four dengue serotypes, of which four significantly inhibited binding of anti-dengue antibodies to the virus. Two peptides with similar sequences to regions of NS3 and NS4B non-structural dengue virus proteins were identified. Conclusion: Our results suggest that these peptides could be used for the development of diagnostic tools for the detection of dengue virus infection and for a potential vaccine against this pathogen.
ABSTRACT
These types of monoclonal antibodies 8E8, 3F7 and 1E9 to dengue 4 virus H-241 strain. These monoclonal antibodies show various patterns of reactivity to the four dengue serotypes and different antigen preparations of serotype 4 when they were tested in various serological methods. The monoclonal antibody 8E8 exhibited a specificity of serotype (D-2; by hemagglutination inhibition); subcomplex (D-2 and D-4 by immunofluorescence) and complex (by immunoperoxidase technique). It was able to neutralize by 80 homologous virus and it turned out to be the only reactive monoclonal antibody in the complement fixation test. The monoclonal 3F7 did not react to by hemagglutination inhibition, recognized serotypes D-1, D-2, D-3 and D-4 by immunofluorescence and only serotypes D1 and D4 by immunoperoxidase technique but it was unable to neutralize the homologous virus. The 1E9 antibody was reactive to serotypes D1, D-2, D-3 and D-4 only by hemagglutination inhibition and neutralized serotype D-4. All the monoclonal antibodies were able to react to various dengue antigens through an ELISA of double antibody and showed fluorescent activity against 38th pass in Beagle dog kidney culture; however, they could not react to a D-4 recombinant antigen.