A broad-spectrum multiple subunit vaccine against Streptococcus suis infection

A streptococcus and vaccine technology, applied in the biological field, can solve the problems of inability to protect multiple serotypes of Streptococcus suis, poor effect, limited antigen expression and exposure, etc.

Active Publication Date: 2020-05-22
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the current commercially available vaccines have reduced the incidence of Streptococcus suis to a certain extent, there are obvious weaknesses, such as the high level of serum antibodies caused by immunization are not all protective antibodies, but the antigen expression or antigens that can provide protective antibodies Exposure is limited, insufficient to induce and provide effective protective effects
In addition, existing research results have confirmed that a single S. suis subunit component cannot provide comprehensive and effective protection against multiple serotypes of S. suis, and commercially available vaccines can only provide cross-protection against a small number of serotypes of S. suis and are not effective

Method used

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  • A broad-spectrum multiple subunit vaccine against Streptococcus suis infection
  • A broad-spectrum multiple subunit vaccine against Streptococcus suis infection
  • A broad-spectrum multiple subunit vaccine against Streptococcus suis infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Embodiment 1, preparation of multiple recombinant protein vaccine V5

[0104] (1) Preparation of recombinant protein

[0105] 1. Preparation of sortase A (SrtA)

[0106] 1. Using the genomic DNA of Streptococcus suis type 2 (A7) as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0107] F1: 5'-CATGCCATGGCCAGCAATGTTACGACAGA-3';

[0108] R1: 5'-CCGCTCGAGTTGTCCATAATCATACTGAT-3'.

[0109] 2. Use restriction endonucleases NcoI and XhoI to double digest the PCR amplification product in step 1, and recover the digested product.

[0110] 3. Digest the vector pET28a(+) with restriction endonucleases NcoI and XhoI, and recover the vector backbone of about 5400bp.

[0111] 4. Ligate the digested product of step 2 with the vector backbone of step 3 to obtain the recombinant plasmid pET28a-SrtA. According to the sequencing results, the results of the recombinant plasmid pET28a-SrtA are described as follows: ...

Embodiment 2

[0173] Example 2, Nasal Immunization V5 Induces Th17 Cell Response

[0174] Four-week-old female C57BL / 6J mice were randomly divided into three groups, and the groups were treated as follows:

[0175] PBS group: On the 0th day, the 7th day and the 14th day of the experiment, 10 μl of PBS buffer was instilled through the nasal cavity, and each mouse was given 10 μl each time.

[0176]V5 group: on the 0th day, the 7th day and the 14th day of the experiment, the V5 vaccine solution was instilled through the nasal cavity respectively, and each mouse was given the V5 vaccine solution each time (the V5 vaccine solution was composed of 10 μg, 10 μg CpG and 10 μg of the five recombinant proteins respectively). 10 μl of PBS buffer obtained by mixing).

[0177] KQ group: on the 0th day, the 7th day and the 14th day of the experiment, the commercially available pre-inactivated vaccine (KQ for short) was injected intramuscularly, and the concentration of the bacterial solution was 1.5×10...

Embodiment 3

[0180] Example 3, V5 immunization induced antibody response in mice

[0181] Four-week-old female C57BL / 6 mice were randomly divided into three groups, and the groups were treated as follows:

[0182] PBS nasal drop group: on the 0th day, 7th day and 14th day of the experiment, PBS buffer was instilled through the nasal cavity respectively, and each mouse was given 10 μl each time.

[0183] V5 group: on the 0th day, the 7th day and the 14th day of the experiment, the V5 vaccine solution was instilled through the nasal cavity respectively, and each mouse was given the V5 vaccine solution each time (the V5 vaccine solution was composed of 10 μg, 10 μg CpG and 10 μg of the five recombinant proteins respectively). 10 μl of PBS buffer obtained by mixing).

[0184] KQ group: on the 0th day, the 7th day and the 14th day of the experiment, the commercially available pre-inactivated vaccine (KQ for short) was injected intramuscularly, and the concentration of the bacterial solution wa...

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Abstract

The present invention discloses a vaccine for preventing Streptococcus suis infection. The active components of the vaccine comprise a component A, a component B, a component C, a component D and a component E; the component A is a sortase or a fusion protein containing the sortase; the component B is SSPA or a fusion protein containing the SSPA; the component C is MRP or a fusion protein containing the MRP; the component D is a SCPC or a fusion protein containing the SCPC; the component E is SLY or a fusion protein containing the SLY; and the vaccine further comprises an immunoadjuvant, and the immunoadjuvant is a mucosal immunoadjuvant CpG or other mucosal immunoadjuvants. The vaccine provided by the invention has the advantages of high efficiency, broad spectrum and low cost. The vaccine adopts a mucosal immunization way, so the vaccine has the characteristics of no tissue damages, no local side effects, simplicity in use, and easiness in promotion and use.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a broad-spectrum multi-unit subunit vaccine for preventing Streptococcus suis infection. Background technique [0002] Streptococcus suis (S. suis) is the pathogenic bacterium that causes streptococcus suis. According to the capsular polysaccharide (CPS) on the surface of the bacteria, Streptococcus suis can be divided into 35 serotypes (1-34 and 1 / 2), Among them, Streptococcus suis type 2 (SS2) is the most virulent and widespread. SS2 is a zoonotic pathogen that can cause diseases such as pig meningitis, encephalitis, pneumonia, endocarditis, polyserositis, arthritis, sepsis and abortion; human infection with SS2 can cause meningitis, sepsis Shock, permanent deafness, endocarditis and even death. Streptococcus suis has brought serious economic losses to the pig breeding and processing industries all over the world. At the same time, it has caused the risk of infection ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/09A61K39/39A61P31/04A61P37/04
CPCA61K39/092A61K39/39A61K2039/543A61K2039/55561A61K2039/57A61K2039/575A61K2039/70A61P31/04A61P37/04
Inventor 王北难邢新新毕帅
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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