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HSV-2 DNA vaccine used by mucous membranes, as well as preparation method and application thereof

An HSV-2DNA, mucosal technology, applied in the fields of genetic engineering and immunology, can solve the problems of no ideal promotion method for vaccines, cumbersome operation, inconvenient use, etc., and achieve the effect of easy promotion and use, simple operation and low cost

Inactive Publication Date: 2017-05-24
WUXI NO 5 PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Invention patent CN 103864902 B discloses a bivalent DNA vaccine comprising HSV-2 gD or gB envelope protein nucleotide sequence and adjuvant CCL19 or CCL28 nucleotide sequence. The immunization route is intramuscular injection. Media to mediate the transfection of DNA plasmids into cells after intramuscular injection may require the use of special equipment, which is cumbersome and inconvenient to use. Therefore, there is currently no ideal way to promote this vaccine clinically, which limits the Application of HSV-2 DNA Vaccine
In addition, this kind of intramuscular DNA vaccine does not take full advantage of the particularity of the HSV-2 infection route, that is, mucosal infection, and its humoral immunity, especially cellular immunity and mucosal immunity mobilization and immune effects need to be further enhanced and improved

Method used

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  • HSV-2 DNA vaccine used by mucous membranes, as well as preparation method and application thereof
  • HSV-2 DNA vaccine used by mucous membranes, as well as preparation method and application thereof
  • HSV-2 DNA vaccine used by mucous membranes, as well as preparation method and application thereof

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preparation Embodiment 1

[0035] 1. Amplification of the target gene:

[0036] HSV-2 gD was successively constructed on the pcDNA3.1(+) plasmid vector (purchased from Invitrogen, the same below) (the experimental strain of HSV-2 was HG52 strain, purchased from LGC Pharmaceutical Impurity Standard Company, Royal Laboratory of England) (GenBank : Z86099.2) envelope glycoprotein gene and adjuvant gene CCL28 (Kai Hu. JOURNAL OF IMMUNOLOGY, Aug. 2013, p.1935-1947), the molecular formulas are: pcDNA3.1(+)-Xba I-gD- PmeI, pcDNA3.1 (+)-HindIII-CCL28-KpnI, named after pgD (for containing the pcDNA3.1 (+) plasmid of the nucleotide sequence shown in SEQ ID NO.1) and pCCL28 (for containing the nucleotide sequence shown in SEQ ID NO.1) respectively pcDNA3.1 (+) plasmid with the nucleotide sequence shown in NO.2), and the primer sequences for amplifying the target fragment are shown in Table 1.

[0037] Table 1 constructs the primer sequence list of pgD and pCCL28

[0038]

[0039] 2. Plasmid Construction

[0...

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Abstract

The invention provides an HSV-2 DNA vaccine used by mucous membranes. The HSV-2 DNA vaccine comprises pgD, pCCL28 and immune mediators, wherein the pgD is a pcDNA3.1(+) plasmid including a nucleotide sequence of HSV-2 gD envelope glycoprotein; the pCCL28 is a pcDNA3.1(+) plasmid including a nucleotide sequence of CCL28; and the vaccine is immunized or used by the mucous membranes. The invention also provides a preparation method of the vaccine. In addition, the invention provides an application of the vaccine in preventing and treating herpes simplex virus infection and treating diseases related to the herpes simplex virus. The HSV-2 DNA vaccine provided by the invention has an ideal immune protection effect and HSV-2 control and disease treatment effects, is convenient in promotion and use, easy in quality control and large-scale production, storage and transportation, low in cost and simple in operation, and has good market application prospects.

Description

technical field [0001] The invention belongs to the field of genetic engineering and immunology, in particular to a HSV-2 DNA vaccine, more specifically, to a HSV-2 DNA vaccine used for mucosal immunity, a preparation method of the vaccine and the preventive properties of the vaccine and therapeutic applications. Background technique [0002] Herpes simplex virus type 2 (HSV-2) is the main pathogen of genital herpes, and there is no safe and effective preventive vaccine available so far. Epidemiological and etiological survey data show that the annual incidence of genital herpes in the world is about 23 million, and HSV-2 infection can increase the chance of HIV-1 infection by at least 3 times, and can accelerate the process of AIDS. What's more serious is that HSV-2 infection will lead to lifelong latent infection and recurrent infection, resulting in heavy economic burden and extensive social and psychological impact. After more than 20 years of basic research and clinic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/245A61K9/00A61K39/39A61K47/36A61P31/22
CPCA61K9/0034A61K9/0043A61K39/12A61K39/39A61K47/36A61K2039/55516A61K2039/575C12N2710/16634
Inventor 阎岩黄利华陈仁芳任勇
Owner WUXI NO 5 PEOPLES HOSPITAL
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