Respiratory syncytial virus vaccine, and preparation method and application thereof

A technology of syncytial virus and respiratory tract, which is applied in the field of respiratory syncytial virus vaccine and its preparation, can solve the problems of low neutralizing antibody level, unguaranteed safety, and the advent of RSV vaccine, so as to inhibit inflammation-related pathological reactions and be easy to promote , effective and specific inflammation-related pathological responses

Active Publication Date: 2016-10-05
ADVACCINE SUZHOU BIOPHARMACEUTICALS CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many types of RSV vaccines that have entered clinical research: (1) developed inactivated vaccines that can produce antibodies against viral proteins, but the level of neutralizing antibodies is low and in the lungs of those who take the vaccines, they are mediated by Th2 and granulocytes. Severe inflammatory cell infiltration, leading to severe disease and even death in the vaccinators after exposure to the virus; (2) subunit vaccines, RSV virus coat proteins F and G are mainly proteins with neutralizing epitopes, in animal models It has been found that the use of purified F and G proteins as vaccines can effectively produce neutralizing antibodies to inhibit virus replication and inhibit the activation of Th2 cells. However, there have been cases of aggravated virus infections after human immunization, and the safety cannot be guaranteed. The subunit vaccine based on RSV F protein has good inhibitory effect on pathological immune response but poor immunogenicity; (3) live attenuated vaccine that can stimulate local and systemic immune response through nasal administration, most of Attenuated vaccines including the most widely used "cpts248 / 404" have a good effect on infants older than 6 months, but moderate inflammatory reactions in the lungs of infants younger than 6 months, and the same safety cannot be guaranteed
In addition, there are many RSV vaccines in clinical research, but so far there is still no approved effective and safe RSV vaccine in the world

Method used

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  • Respiratory syncytial virus vaccine, and preparation method and application thereof
  • Respiratory syncytial virus vaccine, and preparation method and application thereof
  • Respiratory syncytial virus vaccine, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0057] The preparation method of inactivated vaccine FI-RSV: 10 7 TCID 50 RSV virus (US ATCC, catalog no. VR-26 TM ) was reacted with formaldehyde at 37°C for 72 hours, and purified by high-speed centrifugation at 50,000g for 1 hour.

[0058] Cyclosporin A (CSA) is a product of Taiwan Listed Chemical Pharmaceutical Co., Ltd., with a product catalog number of H10940111.

[0059] Vector pET28a(+) is a product of Novagen Company, catalog number 69864-3.

[0060] Escherichia coli BL21(DE3) is a product of Tiangen Biochemical Technology (Beijing) Co., Ltd., catalog number CB105-02.

[0061] 1640 medium is the product of Maichen Technology Co., Ltd., product number CM10040.

[0062] Goat anti-RSV polyclonal antiserum is a product of Meridian Company in the United States, the product catalog number is B656860G.

[0063] HBV vaccine is a product of Huada Pharmaceutical Co., Ltd.

Embodiment 1

[0064] Embodiment 1, the preparation of respiratory syncytial virus (RSV) vaccine

[0065] 1. Acquisition of G protein

[0066] 1. Acquisition of the gene encoding the G protein

[0067] The coding gene sequence of the respiratory syncytial virus G protein of the present invention is shown in sequence 1 in the sequence listing, and the amino acid sequence of the respiratory syncytial virus G protein is shown in sequence 2 in the sequence listing.

[0068] 2. Construction of expression vector pET28a-noG

[0069] Synthesize the coding gene sequence of the respiratory syncytial virus G protein in step 1 by the method of whole gene synthesis, carry out double enzymes to the coding gene sequence of the respiratory syncytial virus G protein and the carrier pET28a (+) with restriction endonuclease NcoI and XhoI cutting and ligation to obtain a recombinant vector, which was named pET28a-noG. The results of enzyme digestion and identification of the pET28a-noG vector are as follows:...

Embodiment 2

[0101] Embodiment 2, detection of antibody level after respiratory syncytial virus (RSV) vaccine immunization mice

[0102] 1. Vaccine immunization mice in each group

[0103] Experimental materials: 6-8 weeks old female BALB / c mice, purchased from the Experimental Animal Department of Fudan University, clean grade, divided into 9 groups, 5 mice in each group. During the experiment, sterile water and food were used for feeding, and the photoperiod was 12 hours.

[0104] Experimental vaccine is divided into the respiratory syncytial virus (RSV) vaccine group (G+CSA), subunit vaccine group (G), PBS group (negative control), FI-RSV group (positive control) of the four preparations of embodiment 1. Respiratory syncytial virus (RSV) vaccine group is divided into (1ugCSA+1ug G protein) group, (2ug CSA+10ug G protein) group, (4ug CSA+10ug G protein) group, (6ug CSA+10ug G protein) group according to CSA and G protein quality difference again in respiratory syncytial virus (RSV) vacc...

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Abstract

The invention discloses a respiratory syncytial virus vaccine, and a preparation method and an application thereof. The active components of the respiratory syncytial virus vaccine comprise a respiratory syncytial virus G protein and an immunosuppressant; the amino acid sequence of the respiratory syncytial virus G protein is represented by sequence 2 in a sequence table; and the immunosuppressant is cyclosporins A. Test proves that the respiratory syncytial virus vaccine can enhance the humoral immunoreaction in the virus neutralizing antibody level of immune animals, can inhibit too strong cellular immunity reactions, effectively and specifically inhibits inflammation related pathological reactions, and has the advantages of manure technology, low cost, no side effects and easy promotion.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a respiratory syncytial virus vaccine and its preparation method and application. Background technique [0002] Respiratory syncytial virus (RSV) is a single-stranded negative-sense RNA virus of the Paramyxoviridae family. It mainly causes lower respiratory tract diseases in newborns and the elderly, and is also one of the main causes of pneumonia in infants under 1 year old. Epidemiological data show that about 60% of newborns are infected with the virus during the RVS epidemic season, and about 18% of children under the age of 5 have been infected with RSV virus, of which about 18% are severe cases and about 20% are hospitalized cases. 15% of outpatient cases. Among those infected, about 30,000 infants and young children die from RSV infection every year worldwide. About 1 / 13 children under the age of five seek medical treatment for RSV every year, and 60% of outpatie...

Claims

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Application Information

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IPC IPC(8): A61K39/155A61P31/14A61P37/02A61K38/13A61K31/436A61K31/573A61K31/675
Inventor 俞庆龄钟一维何忠淮王宾李超凡周娴
Owner ADVACCINE SUZHOU BIOPHARMACEUTICALS CO LTD
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