A vaccine that suppresses streptococcal bacteria and/or prevents streptococcal infections

A type A streptococcus and vaccine technology, which is applied in the direction of antibacterial drugs, bacterial antigen components, medical preparations containing active ingredients, etc., can solve problems that hinder the development of streptococci vaccines, achieve rapid elimination of pathogenic bacteria, and be easy to promote Effect of using and preventing the colonization of pathogenic bacteria

Active Publication Date: 2017-04-05
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These have seriously hindered the development of streptococcal vaccines

Method used

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  • A vaccine that suppresses streptococcal bacteria and/or prevents streptococcal infections
  • A vaccine that suppresses streptococcal bacteria and/or prevents streptococcal infections
  • A vaccine that suppresses streptococcal bacteria and/or prevents streptococcal infections

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, the preparation of sortase A

[0043] 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. The agarose gel electrophoresis of the PCR amplification product is shown in figure 1 .

[0044] F1: 5'-CTTA CATATG GTCTTGCAAGCACAAATGG-3';

[0045] R1: 5'-ATGTT CTCGAG CTAGGTAGATACTTGGTTATAAGA-3'.

[0046] 2. Digest the PCR amplification product of step 1 with restriction endonucleases NdeI and XhoI, and recover the digested product.

[0047] 3. Digest the vector pET28a(+) with restriction endonucleases NdeI and XhoI to recover a vector backbone of about 6000 bp.

[0048] 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 restricti...

Embodiment 2

[0055] Example 2, Immune protection against type A streptococcal infection after nasal cavity inhalation of sortase A and CTB protein

[0056] 1. Female BALB / c mice aged 4-6 weeks were randomly divided into two groups, and the groups were treated as follows:

[0057] PBS group: on the 1st day, 7th day and 14th day of the experiment, PBS buffer was instilled through the nasal cavity;

[0058] Sortase A / CTB 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 sortase A solution prepared in Example 1 and the CTB protein solution, Each mouse was given 20 μg sortase A and 1 μg CTB protein each time).

[0059] On the 21st day of the experiment, the mice were challenged with live bacteria (type A Streptococcus M1, Type A Streptococcus M12, Type A Streptococcus M28 or Type A Streptococcus M49) by intranasal instillation (bacterial solution The c...

Embodiment 3

[0067] Example 3. Mucosal immunization with sortase A and CTB protein induces an immune response dominated by Th17 cells

[0068] Th17 cells play a major role in mucosal immune protection. The purpose of this example is to verify the effect of sortase A and CTB protein on Th17 cell activation.

[0069] Female BALB / c mice aged 4-6 weeks were randomly divided into four groups, and the grouping was handled as follows:

[0070] PBS group: on the 1st day, 7th day and 14th day of the experiment, PBS buffer was instilled through the nasal cavity;

[0071] Sortase A group: On the first day, the seventh day and the 14th day of the experiment, the sortase A solution prepared in Example 1 was instilled through the nasal cavity respectively (each mouse was given 20 μg sortase A each time);

[0072] CTB group: On the first day, the seventh day and the 14th day of the experiment, the CTB protein solution was instilled through the nasal cavity respectively (each mouse was given 1 μg CTB pro...

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Abstract

The invention discloses a vaccine for inhibiting streptococcus and / or preventing streptococcus infection. Active components of the vaccine provided in the invention comprise a component A and a component B; the component A comprises a sortase, a fusion protein containing the sortase, protein formed by conjugated connection of the sortase with adjuvant protein, a connection compound of the sortase and polysaccharide or a DNA expression vector of a coding gene of the sortase; the component B comprises a cholera toxin B subunit and fusion protein containing the cholera toxin B subunit. The vaccine provided in the invention has the superiority of high efficiency, wide spectrum, and low cost. Meanwhile, the vaccine provided in the invention adopts a mucosal immunization approach, has the characteristics of no tissue damage, no local side effect, and simple usage, and is suitable for popularization and application.

Description

technical field [0001] The present invention relates to a vaccine for inhibiting Streptococcus and / or preventing 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, large tissue necrosis occurs within 12-24 hours, leading to death, and the mortality rate is as high as 60%. The lucky ones often end up with amputation at the price. Therefore, they are known as "man-eating bacteria" and may become bioterrorist weapons. Another important reason why Strep A has attracted people's attention is its association with autoimmune diseases. Recurrent streptococcal t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/02A61K39/00A61K39/39A61P31/04
Inventor 王北难范鑫王小爽高斌崔红莲毕帅
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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