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A broad-spectrum multiple subunit vaccine for preventing type a streptococcal infection

A type A streptococcus and vaccine technology, applied in the direction of multivalent vaccines, vaccines, antibacterial drugs, etc., can solve the problems of inability to exert immune protection, and the antibodies cannot be recognized, to improve the level of cell activation and antibodies, and easy to promote Use, no tissue damage effect

Active Publication Date: 2020-01-17
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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  • A broad-spectrum multiple subunit vaccine for preventing type a streptococcal infection
  • A broad-spectrum multiple subunit vaccine for preventing type a streptococcal infection
  • A broad-spectrum multiple subunit vaccine for preventing type a 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, perform PCR amplification 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 restriction endonucleases NdeI and XhoI to double digest the PCR amplified product of step 1, and reclaim the digested product.

[0048] 3. Use restriction endonucleases NdeI and XhoI to double-digest the vector pET28a(+), and reclaim the vector backbone of about 5400bp.

[0049] 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: between the NdeI and XhoI restriction sites of the vector pET28a(+), the sequence 1 of the sequence listing is inserted from 24...

Embodiment 2

[0119] Example 2. Nasal inhalation of Five Comp+CpG promotes the removal of Streptococcus type A in nasal cavity infection sites in mice

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

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

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

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

...

Embodiment 3

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

[0127] Female BALB / c mice aged 4-6 weeks were randomly divided into three groups, and the grouping was handled as follows:

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

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

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

[0131] ...

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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 invention relates to a broad-spectrum multiple subunit vaccine for preventing type A streptococcus infection. Background technique [0002] Type A Streptococcus (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, mild cases include tonsillitis, impetigo, erysipelas and other types In severe cases, severe invasive infections can be life-threatening, such as pneumonia, bacteremia, toxic shock and acute necrotizing fasciitis. In the latter, a large area of ​​tissue necrosis occurs in 12-24 hours and leads to death, 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 takes the first place in the mortality rate of cardiovascular diseases in the worl...

Claims

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Application Information

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Patent Type & Authority Patents(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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