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A combination vaccine for inhibiting and/or preventing type a streptococcal infection

A streptococcus and vaccine technology, applied in the direction of antibacterial drugs, bacterial antigen components, etc., can solve the problems of inability to exert immune protection, hinder vaccine development, and restrict vaccine research and development.

Active Publication Date: 2018-11-27
HAIKOU PHARMA FACTORY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research and development of Streptococcus A vaccine started as early as the 1920s but has not yet come out. Two important reasons have hindered the development of vaccines: First, there are many serotypes of Streptococcus A, and each serotype lacks crossover, making it difficult to develop a vaccine. Vaccine against all serotypes
Streptococcus A is divided into more than 150 serotypes according to the difference of the M protein of its cell wall, and the number is constantly increasing. It is difficult for a vaccine designed for one serotype to work against another serotype, which has become the primary reason for restricting the development of vaccines.
Second, streptococcal infection is closely related to autoimmune diseases, and vaccines have the risk of causing autoimmune diseases, which greatly hinders the process of vaccine development
Although sortase A immunization also induces the production of antibodies, 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 combination vaccine for inhibiting and/or preventing type a streptococcal infection
  • A combination vaccine for inhibiting and/or preventing type a streptococcal infection
  • A combination vaccine for inhibiting and/or preventing type a streptococcal infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1, the preparation of combined vaccine component sortase A and C5a protease

[0062] 1. Preparation of sortase A

[0063] 1. Preparation of recombinant expression vector

[0064] 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 516bp PCR amplification product, which is the sortase A gene SrtA.

[0065] F1: 5'-CTTA CATATG GTCTTGCAAGCACAAATGG-3';

[0066] R1: 5'-ATGTT CTCGAG CTAGGTAGATACTTGGTTATAAGA-3'.

[0067] 2), using restriction endonucleases NdeI and XhoI to double digest the PCR amplified product of step 1, and reclaim the digested product.

[0068] 3) The vector pET28a(+) was double digested with restriction endonucleases NdeI and XhoI, and the vector backbone of about 6000 bp was recovered.

[0069] 4) Ligate the digested product of step 2 with the vector backbone of step 3 to obtain the recombinant plasmid pET28a-SrtA.

[0070] After sequencing, t...

Embodiment 2

[0109] Embodiment 2, the functional research of combined vaccine

[0110] 1. Immune protection of combined vaccine against Strep A bacteria

[0111] The immunogens are grouped as follows:

[0112] Sortase A plus CTB group (SrtA): the sortase A prepared in Example 1 and the adjuvant CTB were mixed at a mass ratio of 10:1 for immunization, and the immunization dose was 10 μg sortase A / time / body;

[0113] C5a protein plus CTB group (SCPA): the C5a protease prepared in Example 1 and the adjuvant CTB were mixed according to the mass ratio of 20:1, and immunized, and the immunization dose was 20 μg C5a protein / time;

[0114] Combined vaccine plus CTB group (SrtA / SCPA): Combined vaccine immunization prepared in Example 1, immunization dose is 20 μg C5a protein / time;

[0115] Streptococcus A full bacteria group (GAS): Type A Streptococcus M1 type immunity, the immune dose is 5×10 7 cfu / time;

[0116] PBS group: use PBS (10mM Na 2 HPO 4 , 1.8mM KH 2 PO 4 , 140mM NaCl, 2.7mM KCl...

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Abstract

The present invention discloses a combined vaccine for inhibiting and / or preventing type A streptococcal infection. The present invention provides a composition for inhibiting streptococcus and / or preventing streptococcal infection, and the composition comprise an immunogen. The immunogen is one of the following 1)-4): 1) the immunogen comprises a sortase A and a C5a protease; 2) the immunogen comprises a sortase A solution and a C5a protease solution; 3) the immunogen comprise a fusion protein containing sortase A and a fusion protein containing C5a protease; and 4) the immunogen comprises a junctional complex of sortase A and polysaccharide and a junctional complex of C5a protease and polysaccharide. The specific amino acid sequence of the sortase A is the N-terminal amino acid residues from site No.22 to No.189 of a sequence 2; and the amino acid sequence of the C5a protease is a sequence 4 in the sequence table. The combined vaccine can be applied to prevention and treatment of mucosal system infections caused a variety of Gram-positive bacteria.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a combined vaccine for inhibiting and / or preventing type A streptococcus infection. Background technique [0002] Group A streptococci (hereinafter referred to as Streptococcus A) is a normal flora of the human upper respiratory tract and one of the common Gram-positive opportunistic pathogens in clinical practice. It can cause a variety of diseases, including Tonsillitis, pharyngitis, and various types of skin and soft tissue infections; severe cases are life-threatening, such as pneumonia, bacteremia, toxic shock syndrome, and acute necrotizing fasciitis. Streptococcus A is ubiquitous in the tonsils of humans. Once it enters the wound, it has a strong reproductive ability in the human body, can quickly produce toxins, erode muscles and body tissues, and cause patients to become disabled, shock, or even die. "Man-eating fungus". Recurrent streptococcal tonsillitis can lead to autoimmun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/116A61K39/09A61P31/04
Inventor 王北难陈宪洋毕帅范鑫王小爽
Owner HAIKOU PHARMA FACTORY
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