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Preparation and application of recombinant adenovirus vector bivalent living vaccine for herpes progenitalis

A recombinant adenovirus, herpes simplex virus technology, applied in the direction of virus/phage, application, antiviral agent, etc., can solve the problems of low response ability, carcinogenic risk, weak immunogenicity, etc., to control recurrence and transmission, high infection ability, the effect of reducing the incidence of

Inactive Publication Date: 2011-06-01
郑州金森生物科技工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective treatment for genital herpes, especially for recurrent genital herpes. Studies have also shown that condoms cannot effectively prevent herpes virus infection. Therefore, the incidence of genital herpes is increasing year by year worldwide. The prevalence has risen to 25%, and it can be said that one in every four adults suffers from genital herpes, so there is an urgent need to develop a vaccine that can effectively prevent and treat genital herpes
[0003] The research results of HSV vaccines in various countries in the world over the past few decades have shown that traditional inactivated vaccines and subunit vaccines have weak immunogenicity and can only induce humoral immune responses, and the effect of inducing cellular immunity is poor; live attenuated vaccines have potential safety hazards and have Cancer risk; although the new DNA vaccine can induce humoral and cellular immunity at the same time, its transfection efficiency is low, and the ability to induce effective mucosal immune response through mucosal immunity is even lower
In the immune response process of herpes simplex virus, cellular immunity and mucosal immunity play a greater role, but the above-mentioned types of vaccines cannot effectively induce cellular immunity and mucosal immune response. Applied vaccines on the market

Method used

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  • Preparation and application of recombinant adenovirus vector bivalent living vaccine for herpes progenitalis
  • Preparation and application of recombinant adenovirus vector bivalent living vaccine for herpes progenitalis
  • Preparation and application of recombinant adenovirus vector bivalent living vaccine for herpes progenitalis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] The preparation of embodiment 1 recombinant adenovirus vector bivalent live vaccine JSA01BD

[0015] 1. Isolation of wild-type HSV-2

[0016] (1). Take the herpes fluid and secretions of patients with clinical HSV infection, add 1ml of PBS buffer solution and filter with a 0.45 micron filter.

[0017] (2). Take a six-well plate, pass 2×10 per well 5 For each Vero cell, 200 microliters of virus filtrate was added to each well of 5 wells, and one well was reserved as a control.

[0018] (3). After 72 hours of incubation in a 37°C carbon dioxide incubator, it was observed that cytopathic changes (CPE) appeared in the wells.

[0019] (4). After the cells in the wells of the six-well plate have complete CPE, put the medium and the cells into the centrifuge tube together.

[0020] (5). Freeze and thaw the centrifuge tube three times at 37°C-minus 80°C, centrifuge at 5000 rpm for 10 minutes, transfer the supernatant into a 2ml EP tube, and put it in a minus 80°C refrigerato...

Embodiment 2

[0126] Embodiment 2 Live vaccine JSA01BD is to the immune protection experiment of guinea pig genital herpes

[0127] 1. Experimental materials: wild-type herpes simplex virus type II: the titer was measured after amplifying HSV-2 with Vero cells, and the titer was about 1×107IU / ml; the titer of recombinant adenovirus padred and JSA01BD was 2×109IU respectively / ml and 1×10 9 IU / ml; 30 common-grade guinea pigs, female, weighing 275±25g, each of which is marked on different parts of its body, and the corresponding marks are converted into 10 Arabic numerals from 1 to 30, according to random According to the chemical principle, 10 rats / group were divided into three groups: blank control group, negative control group and positive group.

[0128] 2. Immunization and attack plan

[0129] Immunity dose: 10 8 TCID50 recombinant adenovirus; route: intranasal instillation, primary immunization at week 0, booster immunization after 2 weeks (d14), and challenge with wild-type HSV-2 af...

Embodiment 3

[0153] Embodiment 3 Live vaccine JSA01BD is to the immune protection experiment of mouse

[0154] 1. Experimental materials: wild-type herpes simplex virus type II: the titer was measured after amplifying HSV-2 with Vero cells, and the titer was about 1×107IU / ml; the titer of recombinant adenovirus padred and JSA01BD was 2×109IU respectively / ml and 1×10 9 IU / ml; ordinary level experimental mice, 30, female, weighing 20±2g, each of which is marked on different parts of its body, after converting the corresponding marks into 10 Arabic numerals from 1 to 30, press According to the principle of randomization, 10 rats / group were divided into three groups: blank control group, negative control group and positive group.

[0155] 2. Immunization and attack plan

[0156] Immunity dose: 10 8 TCID50 recombinant adenovirus; route: intranasal instillation, primary immunization at week 0, booster immunization after 2 weeks (d14), and challenge with wild-type HSV-2 after another two week...

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Abstract

The invention belongs to the technical field of biomedicines, and in particular relates to a novel recombinant adenovirus vector bivalent living vaccine for resisting herpes progenitalis caused by herpes simplex virus type 2 infection. By a molecular biology technology, an expression kit for full-length sequences of genes containing herpes simplex virus type 2 glycoprotein B and glycoprotein D, and an expression kit for cell factor interleukin 2 are cloned to an adenovirus vector to form the vaccine. The vaccine is a replication-defective adenovirus vector living vaccine, only can express protective antigens without pathopoiesia after entering a human body, has high safety, and can simulate the wild herpes virus infection characteristic, effectively induce organism mucosal and humoral immune response and cell immune response, reduce the incidence rate of people, remove viruses which have infected people, and control the relapse and spread of herpes.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a novel recombinant adenovirus vector bivalent live vaccine against genital herpes caused by type 2 herpes simplex virus infection. Background technique [0002] Genital herpes is a viral sexually transmitted disease second only to AIDS, and its pathogen is mainly herpes simplex virus type 2, accounting for more than 90% of primary infections. II herpes simplex virus mainly causes primary and recurrent herpes in the reproductive system after humans, and can remain latent in the ganglion for life and recur when the body's immunity to infection decreases. Pregnant women are prone to miscarriage after being infected with HSV-II virus, and can also cause congenital malformation or mental retardation of the fetus. Infection in newborns during delivery can lead to high fever, dyspnea, or central nervous system lesions, and 60%-70% of newborns may die. In addition, stu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/245A61K39/39A61K48/00C12N7/01C12N15/861A61P31/22
Inventor 刘景伟张利娟胡军
Owner 郑州金森生物科技工程有限公司
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