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Broad-spectrum multi-subunit vaccine for preventing A type streptococcal infection

A vaccine and polysaccharide technology, applied in the direction of multivalent vaccines, vaccines, antibacterial drugs, etc., can solve the problems of inability to exert immune protection and unrecognized antibodies, improve the level of cell activation and antibodies, and be easy to promote and use. The effect of pathogen colonization

Active Publication Date: 2018-02-27
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 sortase A immunization can also induce antibody production, because it is located in the cell wall, antibodies cannot recognize it and thus cannot exert immune protection

Method used

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  • Broad-spectrum multi-subunit vaccine for preventing A type streptococcal infection
  • Broad-spectrum multi-subunit vaccine for preventing A type streptococcal infection
  • Broad-spectrum multi-subunit vaccine for preventing A type streptococcal infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Preparation of Sortase A (SrtA)

[0044] 1. Using the genomic DNA of type A streptococcus M1 as a template, PCR amplification is performed with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0045] F1: 5'-CTTACATATGGTCTTGCAAGCACAAATGG-3';

[0046] R1: 5'-ATGTTCTCGAGCTAGGTAGATACTTGGTTATAAGA-3'.

[0047] 2. Use the restriction enzymes NdeI and XhoI to double-digest the PCR amplification product of step 1, and recover the enzyme-digested product.

[0048] 3. The vector pET28a(+) was double digested with restriction enzymes NdeI and XhoI, and the vector backbone of about 5400bp was recovered.

[0049] 4. Connect the enzyme-digested product of step 2 and the vector backbone of step 3 to obtain a recombinant plasmid pET28a-SrtA. According to the sequencing results, the results of the recombinant plasmid pET28a-SrtA are described as follows: Between the NdeI and XhoI restriction sites of the vector pET28a(+), the sequence 1 of the sequence list...

Embodiment 2

[0119] Example 2. Inhalation of Five Comp+CpG in the nasal cavity promotes the removal of streptococcus type A from the nasal cavity infection site in mice

[0120] 1. Female BALB / c mice aged 4-6 weeks were randomly divided into three groups, and the grouping treatment was as follows:

[0121] PBS group: PBS buffer was instilled through the nasal cavity on the 1st, 7th and 14th days of the experiment respectively;

[0122]Five Comp+CpG group: on the 1st day, the 7th day and the 14th day of the experiment, the vaccine liquid was instilled into the nasal cavity respectively (the vaccine liquid was obtained by mixing the five recombinant proteins prepared in Example 1 and the CpG solution, and each mouse was 10 μg and 10 μg CpG of each of the 5 recombinant proteins were administered each time).

[0123] GAS group: GAS was instilled through the nasal cavity on the 1st, 7th and 14th days of the experiment (the concentration of the bacterial solution was 5 × 10). 7 CFU / 10μl, insti...

Embodiment 3

[0126] Example 3. Nasal inhalation of Five Comp+CpG promotes the immune protection of mice against lethal doses of type A streptococcus.

[0127] Female BALB / c mice aged 4-6 weeks were randomly divided into three groups and treated as follows:

[0128] PBS group: PBS buffer was instilled through the nasal cavity on the 1st, 7th and 14th days of the experiment respectively;

[0129] Five Comp+CpG group: on the 1st day, the 7th day and the 14th day of the experiment, the vaccine liquid was instilled into the nasal cavity respectively (the vaccine liquid was obtained by mixing the five recombinant proteins prepared in Example 1 and the CpG solution, and each mouse was 10 μg and 10 μg CpG of each of the 5 recombinant proteins were administered each time).

[0130] GAS group: GAS was instilled through the nasal cavity on the 1st, 7th and 14th days of the experiment (the concentration of the bacterial solution was 5 × 10). 7 CFU / 10μl, instill 10μl per mouse)

[0131] On the 21st ...

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Abstract

The invention discloses a vaccine for preventing A type streptococcal infection. The vaccine provided by the invention has the active ingredients such as ingredient A, ingredient B, ingredient C, ingredient D, ingredient E and ingredient F; the ingredient A is sortase or fusion protein with sortase; the ingredient B is SCPA or fusion protein with SCPA; the ingredient C is Spy0269 or fusion proteinwith Spy0269; the ingredient D is SCPC or fusion protein with SCPC; the ingredient E is SLO or fusion protein with SLO; and the ingredient F is adjuvant CpG or other mucosal adjuvants. The vaccine provided by the invention has the advantages of high efficiency, broad spectrum and low cost. Meanwhile, the vaccine provided by the invention adopts the way of mucosal immunity, has the characteristicsof no tissue damage, no local side effect and simple and convenient use, and is easy to popularize and use.

Description

technical field [0001] The present invention relates to a broad-spectrum multi-unit subunit vaccine for preventing type A streptococcus infection. Background technique [0002] Group A Streptococcus pyogenes (hereinafter referred to as A Streptococcus), also known as Group A Streptococcus or Group A Streptococcus, can cause a variety of diseases, including tonsillitis, impetigo, erysipelas, etc. Skin and soft tissue infections in severe cases, severe invasive infections, can be life-threatening, such as pneumonia, bacteremia, toxic shock and acute necrotizing fasciitis (Necrotizing Fasciitis). In the latter, extensive tissue necrosis leads to death within 12-24 hours, and the mortality rate is as high as 60%. Repeated infection with streptococcus A leads to autoimmune sequelae, such as acute rheumatic fever, rheumatic heart disease, glomerulonephritis, and psoriasis. Among them, rheumatic heart disease ranks first in the mortality rate of cardiovascular disease in the worl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/09A61P31/04
CPCA61K39/092A61K2039/55561A61K2039/70
Inventor 王北难毕帅
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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