Use of Recombinant E Protein Domain III-Based Enzyme-Linked Immunosorbent Assays for Differentiation of Tick-Borne Encephalitis Serocomplex Flaviviruses from Mosquito-Borne Flaviviruses

ABSTRACT The yellow fever virus (YFV) 17D strain is one of the most effective live vaccines for human use, but the in vivo mechanisms for virulence attenuation of the vaccine and the corresponding molecular determinants remain elusive. The vaccine differs phenotypically from wild-type YFV by the loss of viscerotropism, despite replicative fitness in cell culture, and genetically by 20 amino acid changes predominantly located in the envelope (E) protein. We show that three residues in E protein domain III inhibit spread of 17D in extraneural tissues and attenuate virulence in type I/II interferon-deficient mice. One of these residues (Arg380) is a dominant glycosaminoglycan-binding determinant, which mainly accounts for more rapid in vivo clearance of 17D from the bloodstream in comparison to 17D-derived variants with wild-type-like E protein. While other mutations will account for loss of neurotropism and phenotypic stability, the described impact of E protein domain III changes on virus dissemination and virulence is the first rational explanation for the safety of the 17D vaccine in humans.

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Immunogenicity and Protective Activity of a Chimeric Protein Based on the Domain III of the Tick-Borne Encephalitis Virus E Protein and the OmpF Porin of Yersinia pseudotuberculosis Incorporated into the TI-Complex

International Journal of Molecular Sciences ◽

10.3390/ijms19102988 ◽

2018 ◽

Vol 19 (10) ◽

pp. 2988

Author(s):

Nina Sanina ◽

Natalia Chopenko ◽

Andrey Mazeika ◽

Ludmila Davydova ◽

Galina Leonova ◽

...

Keyword(s):

Yersinia Pseudotuberculosis ◽

Chimeric Protein ◽

Encephalitis Virus ◽

Protective Activity ◽

Ompf Porin ◽

E Protein ◽

Tick Borne Encephalitis ◽

Tick Borne Encephalitis Virus ◽

Domain Iii ◽

Ti Complexes

Tick-borne encephalitis (TBE) is a widespread, dangerous infection. Unfortunately, all attempts to create safe anti-TBE subunit vaccines are still unsuccessful due to their low immunogenicity. The goal of the present work was to investigate the immunogenicity of a recombinant chimeric protein created by the fusion of the EIII protein, comprising domain III and a stem region of the tick-borne encephalitis virus (TBEV) E protein, and the OmpF porin of Yersinia pseudotuberculosis (OmpF-EIII). Adjuvanted antigen delivery systems, the tubular immunostimulating complexes (TI-complexes) based on the monogalactosyldiacylglycerol from different marine macrophytes, were used to enhance the immunogenicity of OmpF-EIII. Also, the chimeric protein incorporated into the most effective TI-complex was used to study its protective activity. The content of anti-OmpF-EIII antibodies was estimated in mice blood serum by enzyme-linked immunosorbent assay (ELISA). To study protective activity, previously immunized mice were infected with TBEV strain Dal’negorsk (GenBank ID: FJ402886). The animal survival was monitored daily for 21 days. OmpF-EIII incorporated into the TI-complexes induced about a 30–60- and 5–10-fold increase in the production of anti-OmpF-EIII and anti-EIII antibodies, respectively, in comparison with the effect of an individual OmpF-EIII. The most effective vaccine construction provided 60% protection. Despite the dramatic effect on the specific antibody titer, the studied TI-complex did not provide a statistically significant increase in the protection of OmpF-EIII protein. However, our results provide the basis of the future search for approaches to design and optimize the anti-TBEV vaccine based on the OmpF-EIII protein.

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Cross protection against lethal West Nile virus challenge in mice immunized with recombinant E protein domain III of Japanese encephalitis virus

Immunology Letters ◽

10.1016/j.imlet.2011.04.003 ◽

2011 ◽

Vol 138 (2) ◽

pp. 156-160 ◽

Cited By ~ 8

Author(s):

Shi-Hua Li ◽

Xiao-Feng Li ◽

Hui Zhao ◽

Tao Jiang ◽

Yong-Qiang Deng ◽

...

Keyword(s):

West Nile Virus ◽

Japanese Encephalitis Virus ◽

Japanese Encephalitis ◽

Encephalitis Virus ◽

Cross Protection ◽

Protein Domain ◽

West Nile ◽

Virus Challenge ◽

E Protein ◽

Domain Iii

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Dengue E Protein Domain III-Based DNA Immunisation Induces Strong Antibody Responses to All Four Viral Serotypes

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0003947 ◽

2015 ◽

Vol 9 (7) ◽

pp. e0003947 ◽

Cited By ~ 38

Author(s):

Monica Poggianella ◽

José L. Slon Campos ◽

Kuan Rong Chan ◽

Hwee Cheng Tan ◽

Marco Bestagno ◽

...

Keyword(s):

Antibody Responses ◽

Protein Domain ◽

E Protein ◽

Domain Iii

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Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes

Biomolecules ◽

10.3390/biom8030082 ◽

2018 ◽

Vol 8 (3) ◽

pp. 82 ◽

Cited By ~ 1

Author(s):

Vasily Golotin ◽

Nina Sanina ◽

Ludmila Davydova ◽

Natalia Chopenko ◽

Andrey Mazeika ◽

...

Keyword(s):

Conformational Changes ◽

Yersinia Pseudotuberculosis ◽

Chimeric Protein ◽

Encephalitis Virus ◽

Protein Domain ◽

Scanning Calorimetry ◽

Metal Affinity ◽

Tick Borne Encephalitis ◽

Tick Borne Encephalitis Virus ◽

Domain Iii

Domain III (DIII) of the tick-borne encephalitis virus (TBEV) protein E contains epitopes, which induce antibodies capable of neutralizing the virus. To enhance the immunogenicity of this protein, which has a low molecular weight, the aim of the present work was to express, isolate, and characterize a chimeric protein based on the fusion of the bacterial chaperone HSP70 of Yersinia pseudotuberculosis and EIII (DIII + stem) as a prospective antigen for an adjuvanted delivery system, the tubular immunostimulating complex (TI-complex). The chimeric construction was obtained using pET-40b(+) vector by ligating the respective genes. The resulting plasmid was transformed into DE3 cells for the heterologous expression of the chimeric protein, which was purified by immobilized metal affinity chromatography (IMAC). ELISA, differential scanning calorimetry, intrinsic fluorescence, and computational analysis were applied for the characterization of the immunogenicity and conformation of the chimeric protein. Mice immunization showed that the chimeric protein induced twice the number of anti-EIII antibodies in comparison with EIII alone. In turn, the incorporation of the HSP70/EIII chimeric protein in the TI-complex resulted in a twofold increase in its immunogenicity. The formation of this vaccine construction was accompanied by significant conformational changes in the chimeric protein. Using HSP70 in the content of the chimeric protein represents an efficient means for presenting the main antigenic domain of the TBEV envelope protein to the immune system, whereas the incorporation of this chimeric protein into the TI-complex further contributes to the development of a stronger immune response against the TBEV infection.

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