Fusion protein vaccine capable of inhibiting Streptococcus and/or preventing Streptococcus infection

A fusion protein and protein technology, applied in the direction of bacteria, hybrid peptides, fungi, etc., can solve problems that hinder the development of type A streptococcal vaccines

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

AI Technical Summary

Problems solved by technology

[0004] Due to the large number of serotypes of Streptococcus A and the fact that they can cause autoimmune diseases, the development of Streptococcus A vaccines has been seriously hindered. As a result, there is currently no vaccine against this bacteria available in the world. Therefore, the development of safe, effective, broad-spectrum A Strep A vaccine is urgently needed

Method used

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  • Fusion protein vaccine capable of inhibiting Streptococcus and/or preventing Streptococcus infection
  • Fusion protein vaccine capable of inhibiting Streptococcus and/or preventing Streptococcus infection
  • Fusion protein vaccine capable of inhibiting Streptococcus and/or preventing Streptococcus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Example 1. Construction of CTB-Sortase A protein gene and preparation of CTB-Sortase A protein

[0115]1. Replace the DNA fragment between the NcoI and BamHI recognition sequences of the vector pET26b(+) with the DNA sequence shown in positions 112-996 of SEQ ID No.2, keep other DNA sequences unchanged, and obtain the recombinant expression vector pET26b- CTB-Sortase A. Sequencing results showed that the DNA sequence shown in positions 112-996 of SEQ ID No.2 was inserted into the recombinant expression vector pET26b-CTB-Sortase A, and the recombinant expression vector pET26b-CTB-Sortase A could express the first sequence of SEQ ID No.1. The CTB-Sortase A protein shown at positions 23-314, the DNA sequence shown at positions 115-996 of SEQ ID No.2 is the CTB-Sortase A protein shown at positions 23-314 of SEQ ID No.1. coding sequence.

[0116] 2. Transform the recombinant vector pET26b-CTB-Sortase A into BL21(DE3)plysS competent cells, and obtain the 23rd code containin...

Embodiment 2

[0124] Example 2. Immune protection against type A streptococcus infection after nasal inhalation of CTB-Sortase A protein

[0125] Female BALB / c mice aged 6-8 weeks were randomly divided into three groups, 6 mice in each group, and the grouping treatment was as follows:

[0126] PBS group: 10 μL of PBS buffer was instilled through the nasal cavity on the first day, the fifth day, the 10th day and the 14th day of the experiment respectively;

[0127] CTB-Sortase A protein 5μg group: 10 μL CTB-Sortase A protein solution (vaccine liquid) was instilled through the nasal cavity on the 1st day, the 5th day, the 10th day and the 14th day of the experiment respectively. 5μg CTB-Sortase A protein;

[0128] CTB-Sortase A protein 10 μg group: 10 μL CTB-Sortase A protein solution (vaccine liquid) was instilled through the nasal cavity on the 1st day, the 5th day, the 10th day and the 14th day of the experiment respectively. is 10μg CTB-Sortase A protein;

[0129] On the 21st day of th...

Embodiment 3

[0131] Example 3. Immune protection against type A streptococcus infection after nasal inhalation of different doses of CTB-Sortase A protein

[0132] Female BALB / c mice aged 6-8 weeks were randomly divided into four groups, 6 mice in each group, and the grouping treatments were as follows:

[0133] PBS group: 10 μL of PBS buffer was instilled through the nasal cavity on the first day, the fifth day, the 10th day and the 14th day of the experiment respectively;

[0134] CTB-Sortase A protein 1 μg group: 10 μL of CTB-Sortase A protein solution (vaccine liquid) was instilled through the nasal cavity on the 1st day, the 5th day, the 10th day and the 14th day of the experiment respectively. is 1 μg CTB-Sortase A protein;

[0135] CTB-Sortase A protein 2.5 μg group: 10 μL CTB-Sortase A protein solution (vaccine liquid) was instilled through the nasal cavity on the 1st day, the 5th day, the 10th day and the 14th day of the experiment respectively. Both were 2.5μg CTB-Sortase A pro...

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Abstract

The invention discloses a fusion protein vaccine capable of inhibiting Streptococcus and/or preventing Streptococcus infection. With fusion protein obtained through linkage of sortase A and a cholera toxin B subunit through connecting peptide, the Th17 cell activation level is remarkably increased, the effects of preventing pathogenic bacteria from settling and quickly removing the pathogenic bacteria can be realized, and the fusion protein has protecting effects on Group A Streptococcus of different serotypes and has the superiority of high efficiency, broad spectrum and low cost. With the fusion protein, the use amount of immunogen is reduced, the production technology is simplified, and the production cost is reduced. Meanwhile, the vaccine adopts the way of mucosal immunity, has the characteristics of being free of tissue damage, free of local side effects and convenient to use, and is easy to popularize and use.

Description

technical field [0001] The present invention relates to a fusion protein vaccine for inhibiting streptococcus and / or preventing streptococcus infection in the field of biotechnology. Background technique [0002] Group A Streptococcus pyogenes is a common clinical Gram-positive opportunistic pathogen, which can cause a variety of diseases, including pharyngitis, impetigo, erysipelas and cellulitis in mild cases, and severe cases can be Life-threatening, such as pneumonia, bacteremia, toxic shock, and acute necrotizing myositis [1] . According to statistics, at least 517,000 people worldwide die of type A streptococcus infection every year, most of them die from rheumatic fever and rheumatic heart disease caused by type A streptococcus infection, and about 163,000 people die from acute type A streptococcus infection. Infect [2] . [0003] There are more than 150 serotypes of Streptococcus A, and new serotypes are constantly emerging. Infection with one serotype does not ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/15C12N1/19C12N1/21C12N1/13A61K39/09A61K39/00A61P31/04
Inventor 王北难毕帅
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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