Zika virus vaccine composition and application thereof

A vaccine composition and virus technology, applied in the field of recombinant proteins, can solve the problems of lack of neutralization of dengue virus, dengue virus infection, etc., and achieve the effect of reducing severe dengue virus infection and improving immunity

Active Publication Date: 2019-08-23
吴夙钦
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The prevalence of Zika virus and dengue virus overlaps, and the anti-Zika virus antibody will cross-react with dengue virus, but lacks the ability to neutralize dengue virus, and may even interact with the Fcγ receptor (Fcγ receptor) to promote dengue virus invasion into cells, resulting in antibody-dependent enhancement (antibody-dependentenhancement, ADE) and more severe symptoms of dengue virus infection, such as hemorrhagic dengue fever (dengue hemorrhagic fever, DHF) with a higher lethality ) and dengue shock syndrome (DSS)

Method used

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  • Zika virus vaccine composition and application thereof
  • Zika virus vaccine composition and application thereof
  • Zika virus vaccine composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Preparation of Recombinant Adenovirus Expressing Envelope Mutants

[0046] In order to shield the Zika virus envelope protein fusion loop and its adjacent regions with cross-reactivity, six kinds of envelope protein mutants with an N-sugar chain modification in the fusion loop region are designed in this embodiment, and its N-glycosyl position as figure 1 shown. Compared with the amino acid sequence (SEQ ID NO: 1) of the wild-type Zika virus envelope protein, the six envelope protein mutants have one or two amino acid substitutions to realize N-glycosylation, including the 74th and The cysteine ​​(cysteine) and proline (proline) at the 75th amino acid position are replaced with asparagine and valine (valine), and the cysteine ​​at the 105th amino acid position is replaced with threonine Replace the two alanines at the 248th and 250th amino acid positions with asparagine and threonine, and replace the arginine at the 252nd amino acid position with asparagine Amino aci...

Embodiment 2

[0056] Antigen-specific IgG elicited by recombinant adenovirus immunization

[0057] In order to test the immune effect of the Zika virus envelope protein mutant of the present invention, the PBS solution (negative control group) or the embodiment of two doses were given to BALB / c mice (4-5 in each group) in the intraperitoneal injection mode. 1 recombinant adenovirus (10 8 PFU / 200μL), and the total IgG titer of anti-Zika virus envelope protein domain 3 (ZDIII for short) in mouse serum was detected by enzyme-linked immunosorbent assay (ELISA). The interval between the second immunization injections was about three weeks, and the blood samples of the mice were collected two weeks after the second injection. The endpoint titer of total IgG is the serum dilution factor that makes the OD450 measurement value reach 0.2 in the ELISA experiment.

[0058] image 3 Shows the total IgG titer of anti-ZDIII recombinant protein in the sera of immunized mice, N.D. in the figure means not...

Embodiment 3

[0060] Immunization with recombinant adenovirus elicits Zika virus neutralizing antibodies

[0061] In this example, virus plaque neutralization test (PRNT) was used to measure the titer of neutralizing antibodies against Zika virus in the immune serum of mice after different immunization injections according to the method described in Example 2. During the experiment, ten-fold diluted immune serum or its two-fold serial dilution was mixed with 100 PFU Zika virus, and then the mixture of serum and virus was used to infect Vero cells to calculate the number of virus plaques and obtain the neutralization curve. The titer of neutralizing antibody is defined as the serum dilution factor that reduces the number of viral plaques by 50%, expressed as PRNT 50 express.

[0062] Figure 4A Shows the neutralization curve of Zika virus neutralizing antibody in sera of immunized mice; Figure 4B displayed by Figure 4A Calculated titers of neutralizing antibodies. in accordance with ...

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Abstract

The invention provides a Zika virus envelope protein mutant and a nucleic acid molecule including a nucleotide sequence encoding the mutant. Preferably, the Zika virus envelope protein mutant has a threonine substitute at the 105th amino acid of a wild type Zika virus envelope protein, or has an asparagine substitute and a threonine substitute at the 248th amino acid and the 250th amino acid respectively, and thus the Zika virus envelope protein mutant has an N-carbohydrate chain shielding the fusion loop zone of the Zika virus envelope protein. The invention also provides a vaccine composition which includes the Zika virus envelope protein mutant or a recombinant virus with the nucleic acid molecule. The invention also provides application of the Zika virus envelope protein mutant in preparing the vaccine composition for preventing Zika virus infection or reducing the antibody-dependent enhancement of dengue virus infection.

Description

technical field [0001] The present invention relates to a recombinant protein, nucleic acid molecule and application thereof, in particular to a Zika virus envelope protein mutant and a nucleic acid molecule having its corresponding nucleotide sequence, a kind of envelope protein mutant or The vaccine composition of the recombinant virus having the nucleic acid molecule, and the use of the mantle protein mutant for preparing the vaccine composition. Background technique [0002] Zika virus (ZIKV), a virus transmitted by Aedes mosquitoes, has become the focus of global public health since its outbreak in Brazil in 2015. According to the statistics of the World Health Organization (World Health Organization) in 2016, the Zika virus has spread to more than 70 countries in tropical and subtropical regions of Asia, North America, Africa, and Australia. More than 2.5 million people worldwide are at risk of Zika virus infection due to the widespread spread of the mosquito vector. ...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C07K14/18C12N15/40A61K39/12A61P31/14
CPCC07K14/005A61K39/12C12N2770/24122C12N2770/24134Y02A50/30
Inventor吴夙钦杨劭苹林晓含
Owner吴夙钦