Vaccine for preventing herpesvirus hominis type II

A herpes simplex virus, vaccine technology, applied in antiviral agents, medical preparations containing active ingredients, gene therapy and other directions, can solve the problems of affecting practical application, low efficiency, etc. The effect of morbidity

Inactive Publication Date: 2015-06-24
ZHEJIANG ACAD OF MEDICAL SCI
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

DNA vaccines can induce humoral immunity and cellular immunity in the immune response, and have been successfully applied to various animals such as mice, sheep, chickens, monkeys and chimpanzees, but their effi

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Vaccine for preventing herpesvirus hominis type II
  • Vaccine for preventing herpesvirus hominis type II
  • Vaccine for preventing herpesvirus hominis type II

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0016] Example 1: Obtaining eukaryotic expression plasmid

[0017] Using molecular biology technology PCR method, restriction enzyme digestion, ligation and cloning screening, the original resistance ampicillin (Amp) on the vector pcDNA3 was replaced with Kan resistance (Kan), and then the eukaryotic expression plasmid vector pcDNA3-Kan was constructed. PCR method amplifies the HSV-2 sav strain glycoprotein D full gene sequence (gD), after cloning and recombination, a recombinant plasmid pcDNA-Kan / gD containing gD is obtained. The recombinant plasmid pcDNA-Kan / gD and its specific preparation method have been compared with "He Fang et al. Chinese Journal of Health Inspection, 2008, 18(6): 997-999".

Example Embodiment

[0018] Example 2: Obtaining recombinant plasmid

[0019] Take activated T cells, extract total RNA with Rneasy Mini Kit (QIAGEN), use Oligod(T)n as primer, reverse transcription to synthesize first strand cDNA. With reference to the BC119225.1 sequence included in the NCBI nucleic acid database, a pair of primers was designed, and the synthesized cDNA was used as a template to amplify SEQ ID NO.1. Upstream primer sequence is 5 ， -GCC GAATTC ATGATAGAAACATACAGCCAACCT-3 ， , The downstream primer sequence is 5 ， -GAC CTCGAG TCAGAGTTTGAGTAAGCCAAAAGA-3 ， .

[0020] The gene sequence shown in SEQ ID NO. 1 and pcDNA3-Kan were digested with EcoRI and Xho I, and then the gel was purified and recovered. The resulting fragments were recovered and ligated under the action of T4 ligase at a molar concentration of 1:5. The ligation product was transformed into a competent strain of DH5α. The next day, a medium-sized white colony was picked and inoculated in Kan-resistant LB culture medium wi...

Example Embodiment

[0022] Example 3: Using the plasmids obtained in Examples 1 and 2 to immunize mice

[0023] 1. Experimental materials:

[0024] 1) The plasmids obtained in Examples 1 and 2 were amplified in E. coli and purified with QIAGEN purification kit. The concentration and purity of plasmid were determined by ultraviolet spectrophotometry. The concentration and purity of DNA were determined by OD260 and OD280. Plasmid DNA with a ratio of OD260 / OD280 between 1.8 and 2.0 was selected to immunize mice. The two plasmids were stored separately at a low temperature of -20°C, and the mice were thawed before immunization, and the two plasmids were mixed according to the mass ratio of 2:1.

[0025] 2) Freeze storage of HSV-2 virus sav3 repeatedly for three times. After centrifugation, the supernatant was added to the Vero cell monolayer and incubated at 37°C for 1 hour. Remove the virus liquid, add 4% FBS DMEM complete medium, and incubate at 35°C for 24-48h. The adherent cells were scraped off wit...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a vaccine for preventing herpesvirus hominis type II. The glycoprotein D gene of the herpesvirus hominis type II is combined into a eukaryotic expression vector pKan with Kanamycin resistance by a molecular biology method, so as to construct a DNA vaccine containing a eukaryotic expression plasmid pgD containing a full-length glycoprotein D gene and a recombinant plasmid containing a gene sequence as shown in SEQ ID NO.1. The body immunized with the DNA vaccine can produce effective humoral immunity and cellular immunity to prevent and treat diseases caused by infection with the herpesvirus hominis type II, for example, herpes progenitalis.

Description

technical field [0001] The invention belongs to a biological product, in particular to a vaccine for preventing herpes simplex virus type II (HSV-2). Background technique [0002] Genital herpes is a sexually transmitted disease that is relatively common in the world and in coastal areas of my country, and its incidence is gradually increasing. It is caused by the human herpes simplex virus (HSV) that infects the skin and mucous membranes of the genitourinary and anus through sexual transmission. Herpes simplex virus can be divided into types 1 and 2. Most genital herpes is caused by HSV-2 infection. The main source of infection is genital herpes patients and asymptomatic virus carriers. HSV-2 infection is lifelong, the virus has neurotropic latency, and the virus can establish lifelong latency in the human sacrococcygeal ganglion. HSV-2 latent infection can lead to recurrence of relevant clinical symptoms due to activation of various non-specific stimuli, such as typical g...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K48/00A61K39/245A61P31/22
Inventor 陶薇洪艳胡如西傅婷何卓晶贾岚
Owner ZHEJIANG ACAD OF MEDICAL SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap