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Vaccine for prevention of respiratory syncytial virus infection

A technology for virus infection and respiratory tract, applied in the direction of antiviral agents, antibody medical ingredients, medical preparations containing active ingredients, etc., can solve the problems of limited protection ability, limited effect, high cost, and achieve easy promotion, mature technology, low cost low effect

Inactive Publication Date: 2017-03-29
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although formalin inactivated vaccines, F or G protein subunit vaccines can induce the body to produce a high level of protective neutralizing antibodies, when the body is re-infected with the virus, they will induce enhanced pathological reactions to varying degrees; attenuated vaccines Due to problems such as genetic instability and application safety, it is not suitable for large-scale promotion and use; although the development and research of new vaccines such as genetic engineering vaccines and nucleic acid vaccines are progressing rapidly, their protective capabilities are limited, and most of them are in the stage of animal experiments. Hard to make a big breakthrough
At this stage, passive immunization is mainly used to prevent and treat RSV infection. However, due to the high cost and limited effect, it is difficult to promote it on a large scale.

Method used

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  • Vaccine for prevention of respiratory syncytial virus infection
  • Vaccine for prevention of respiratory syncytial virus infection
  • Vaccine for prevention of respiratory syncytial virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Embodiment 1, the preparation of subunit vaccine

[0100] 1. Expression vector construction: RSV surface glycoprotein F was obtained by whole gene synthesis after the codon optimization of E. coli cells. The nucleotide sequence (GeneBank: AY911262.1) corresponding to the 412-524th amino acid of RSV F protein was synthesized by whole gene synthesis technology (as shown in sequence 1 in the sequence table), and repeated once, that is, 2 times of F sequence. Before synthesis, the nucleotide sequence was optimized according to the codon usage preference of Escherichia coli. The sequence optimization not only removed rare codons, but also deleted the N-terminal of the F protein, and subcloned the optimized F protein nucleotide sequence To the prokaryotic expression vector pET28a(+), named pET28a-A2Fa*2, wherein NcoI and XhoI were selected as restriction sites (as shown in sequence 2 in the sequence listing), and the size was obtained by agarose gel electrophoresis to be abo...

Embodiment 2

[0113] Example 2, Detection of Neutralizing Antibody Levels After Vaccine Immunization in Model Animals

[0114] 1. RSV F A2Fa*2 subunit vaccine immunization model animal experiments

[0115] Model animals: 6-8 weeks old female clean grade BALB / c mice were purchased from the Experimental Animal Department of Fudan University, and the experiment was divided into 5 groups with 5 mice in each group; sterile water and food were used for feeding during the experiment, and the light cycle was 12 hours ;

[0116] Vaccine and control for immunization: PBS, formalin inactivated vaccine FI-RSV (10 7 TCID 50 RSV virus (US ATCC, catalog no. VR-26 TM , reacted with formaldehyde at 37°C for 72 hours, then purified by high-speed centrifugation at 50,000g for 1 hour), and the F subunit vaccine is the above-mentioned recombinant protein;

[0117] The experimental groups and the immunization status of each group are shown in Table 2. Dissolve the F protein A2Fa*2 lyophilized product in a v...

Embodiment 3

[0129] Example 3, Detection of T lymphocyte expansion after A2Fa*2 vaccine immunization animals

[0130] 1. Experiments on A2Fa*2 subunit vaccine immunization model animals

[0131] Same as Step 1 of Example 2.

[0132] 2. Detection of T lymphocyte expansion in the cellular immune response of combined immunosuppressive RSV F protein A2Fa*2 vaccine

[0133] The mice were sacrificed on the 7th day after the last immunization in the above step 1, and the mouse spleen was taken under aseptic conditions, crushed and removed red blood cells with red blood cell lysate, filtered through a nylon column to remove B cells to make a single cell suspension (T lymphocytes) cells), washed 3 times with PBS, centrifuged and counted cells, and adjusted the cell concentration to 1×10 6 cells / ml, each group of cell suspension was added into 96-well culture plate in 3 parts, and the total number of cells in each well was 4×10 5 indivual. One part (group 1-group 7 mouse cells) plus the antigen ...

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Abstract

Belonging to the field of biological products, the invention relates to a vaccine for prevention of respiratory syncytial virus (RSV) infection caused diseases, and in particular relates to a vaccine that is composed of a subunit vaccine and an immunoadjuvant and is used for prevention of respiratory syncytial virus infection caused diseases. The vaccine is composed of an RSV subunit vaccine, and the subunit vaccine is protein shown as sequence 1 in sequence table (GeneBank:AY911262.1). Experiments prove that the RSV vaccine provided by the invention can enhance the humoral immune response after immunizing animals, also can inhibit too strong cellular immune response, effectively inhibits inflammatory reaction, effectively protects mammalian lung tissues and functions from being destroyed by RSV infection.

Description

technical field [0001] The invention belongs to the field of biological products, and relates to a vaccine for preventing diseases caused by respiratory syncytial virus infection, in particular to a vaccine composed of a subunit vaccine and an immune adjuvant for preventing diseases caused by respiratory syncytial virus infection . Background technique [0002] According to reports, respiratory syncytial virus (RSV) is an important pathogen causing viral lower respiratory tract diseases in infants and young children worldwide. In the United States alone, about 100,000 people are hospitalized each year due to RSV infection, and the resulting cost is about 300 million US dollars. According to the investigation, there were not only sporadic RSV infection but also several large-scale outbreaks in my country. Among 1402 infant patients in Beijing during 2000-2002, 66.1% of the viral respiratory tract infections were caused by RSV. During the period from 1985 to 1992, four pande...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/155A61P31/14
Inventor 王宾钟一维陈璇李超凡
Owner FUDAN UNIV
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