Therapeutic DC compound vaccine against herpes simplex virus and preparation method thereof

A herpes simplex virus, therapeutic technology, applied in the field of vaccines, to achieve the effect of reducing lesions and increasing viral gene transcription and expression

Active Publication Date: 2022-06-21
上海兴瑞一达生物科技有限公司
View PDF9 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

HSV-2 vaccines with envelope proteins such as gD, gB, gH, and gL as antigens can induce HSV-specific neutralizing antibodies, among which gD has the strongest ability to induce neutralizing antibodies, but for effective HSV-2 vaccines, Stimulation with gD is not sufficient and immunization with envelope is not sufficient for effective HSV-2 vaccine

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
  • Therapeutic DC compound vaccine against herpes simplex virus and preparation method thereof
  • Therapeutic DC compound vaccine against herpes simplex virus and preparation method thereof
  • Therapeutic DC compound vaccine against herpes simplex virus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The antigenic gene fragments GM-CSF-gC+US10, GM-CSF-gD+ICP4, and GM-CSF-UL47 were inserted into the lentiviral expression vector pLent-C-GFP respectively, and the corresponding three plasmids were prepared. The gC+US10 The nucleic acid artificial sequence, gD+ICP4 nucleic acid artificial sequence, and UL47 nucleic acid artificial sequence were added with NotI and AsiSI site restriction sites respectively to synthesize a complete expression cassette, and then inserted into the lentiviral pLent-C-GFP vector after restriction enzyme digestion. (Invitrogen) NotI-AsiSI site, transformed into E.coli (Top10), after the correct identification by nucleic acid sequencing, the plasmid was extracted and purified using an endotoxin-free plasmid purification kit to obtain a recombinant expression vector plasmid

[0042] In more detail, this embodiment adopts the following steps:

[0043] The antigenic gene fragments GM-CSF-gC+US10, GM-CSF-gD+ICP4, GM-CSF-UL47 were inserted into the l...

Embodiment 2

[0047] The lentiviral packaging cell line 293T was inoculated in a 10cm culture dish containing DMEM+10% FBS; the reaction system was prepared, mixed well, placed at room temperature for 20 minutes, and then added dropwise to the culture dish containing 293T cells, and then placed in a CO2 incubator Cultured in medium; cultured after transfection to obtain lentiviral virus liquid carrying genes encoding GM-CSF-gC+US10, GM-CSF-gD+ICP4, and GM-CSF-UL47.

[0048] In more detail, this embodiment adopts the following specific steps:

[0049] The Lentiviral Packaging Kit was used. The specific method was as follows: The lentiviral packaging cell line 293T was inoculated into a 10cm culture dish containing DMEM+10% FBS, and cultured at 37°C under 5% CO2 conditions, and the adherence rate was 70%. -80% ready for transfection. Take a sterile 1.5ml EP tube or 15ml centrifuge tube and prepare the reaction system according to the following components: serum-free DMEM: 4ml; pLent-gC+US10,...

Embodiment 3

[0051] Peripheral venous blood was collected from patients with herpes simplex virus, peripheral blood mononuclear cells were isolated, adherent cells were collected, and cytokines were added to induce mononuclear cells to differentiate into DCs, and imDCs were cultured.

[0052] In more detail, this embodiment adopts the following specific steps:

[0053] Collect 50 ml of peripheral venous blood from patients with herpes simplex virus, use TBD sample density separation solution (purchased from Tianjin Haoyang Huake Biotechnology) to separate peripheral blood mononuclear cells, and culture them in 10 ml of medium (purchased from TaKaRa Company, GT-T551), After culturing for 3-4 hours at 37°C and 5% CO2, the suspended T cells were discarded, the adherent cells were collected, and cytokines rhIL-4 (final concentration: 50ng / ml), rhGM-GSF (final concentration: 50ng / ml) were added. 100ng / ml), mononuclear cells were induced to differentiate into DCs, new medium was replaced every 4...

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 therapeutic DC compound vaccine against herpes simplex virus and a preparation method thereof, comprising three kinds of dendritic cells loaded with HSV-2 virus-specific immune antigen gene fragments modified, and the three kinds of HSV-2 virus-specific immune antigen gene fragments loaded with dendritic cells The gene fragments of sexual immune antigens were gC+US10, DC‑UL47, and gD+ICP4, respectively. The present invention uses the granulocyte-macrophage cluster factor receptor signal peptide to guide the expression of the antigen, which can improve the expression efficiency of the antigen, enhance its immune effect, and generate a stronger anti-HSV specific immune response.

Description

technical field [0001] The invention relates to the technical field of vaccines, in particular to a therapeutic DC composite vaccine against herpes simplex virus and a preparation method thereof. Background technique [0002] Herpes simplex virus (HSV) infection is a common infectious disease in humans. The virus is divided into two serotypes, HSV-1 and HSV-2. Among them, HSV-2 infection is one of the most common sexually transmitted diseases, and is the main pathogen of genital herpes and ulcers. It has infected more than 500 million people worldwide, and it is estimated that there are 23 million new infections every year. At present, the clinical treatment of genital herpes is mainly through the use of antiviral drugs to relieve symptoms, but it cannot completely remove the HSV-2 virus and cannot effectively control the recurrence of the disease. [0003] The genome of HSV contains about 85 open reading frames, allowing HSV to produce at least 85 unique proteins. It main...

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
Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/245A61P31/22
Inventor 刘明录王立新刘敏金海锋强邦明张传鹏王亮马洪华冯建海
Owner 上海兴瑞一达生物科技有限公司
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