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Polypeptide with mucosal immunity adjuvant activity, and application of polypeptide in preparation of mucosal immunity adjuvant

A mucosal immune adjuvant and immune adjuvant technology, applied in the fields of immunology and preventive medicine, can solve problems such as safety concerns, large molecular weight, and complex processes, and achieve good clinical application prospects, easy quality control, and human safety high effect

Active Publication Date: 2015-03-11
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, their toxicity limits clinical applications. For example, an influenza vaccine launched in the United States in 2003 used LT as an adjuvant. After vaccination, it was found that it could cause severe side effects such as Bell's palsy, so its use was discontinued.
[0005] In order to avoid toxicity, people try to use the B subunits of CT and LT as adjuvants. However, since both LTB and CTB contain more than 100 amino acids and have relatively large molecular weights, they can only be produced by genetic engineering, and the process is complicated and the preparation cost is high.
Moreover, due to concerns about its safety, the FDA has not yet approved it for clinical use

Method used

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  • Polypeptide with mucosal immunity adjuvant activity, and application of polypeptide in preparation of mucosal immunity adjuvant
  • Polypeptide with mucosal immunity adjuvant activity, and application of polypeptide in preparation of mucosal immunity adjuvant
  • Polypeptide with mucosal immunity adjuvant activity, and application of polypeptide in preparation of mucosal immunity adjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of the polypeptide having mucosal immune adjuvant activity of the present invention

[0021] The amino acid sequence of the polypeptide having mucosal immune adjuvant activity of the present invention is shown in SEQ ID NO: 1, which is synthesized by a solid-phase method, and it is obtained immediately.

[0022] SEQ ID NO: 1: HIDSQKKA.

[0023] After searching by NCBI / BLAST, the polypeptide sequence is completely identical to the 21-28th amino acid sequence of the heat-labile enterotoxin B chain (LTB) of Escherichia coli strain 2788150 and other Escherichia coli (GenBank NO: EMW30130.1), and also with the 45th-52nd positions of the cholera enterotoxin B subunit (cholera enterotoxin B subunit, CTB) of Vibrio cholerae serotype O1biotype El Tor, etc. The amino acid sequences are completely consistent (GenBank NO: AAY43121.1). After PUBMED search, it was found that the sequence is the T cell epitope adjacent to CTB (PMID: 8921943) and the T-cell and ...

experiment example 1

[0048] Experimental example 1 Safety detection of the polypeptide having mucosal immune adjuvant activity of the present invention

[0049] 1. Experimental method

[0050] Using BALB / c mice as an animal model, intraperitoneally inject the polypeptide with mucosal immune adjuvant activity prepared in Example 1 of the present invention in mice, and test its safety:

[0051] a) Six healthy male mice aged 3-4 weeks were obtained from the Experimental Animal Center of Chongqing Medical University.

[0052] b) Dissolving the polypeptide having mucosal immune adjuvant activity prepared in Example 1 with PBS buffer.

[0053] c) Add 0.50ml of the mucous membrane immune adjuvant activity of 1.0μg / ml, 2.0μg / ml, 5.0μg / ml, 10.0μg / ml, 20.0μg / ml, 50.0μg / ml, 100.0μg / ml Polypeptides (without exogenous endotoxin detection and treatment) were intraperitoneally injected into the mice described in a) above, and the differences in signs between the experimental rabbits and the control group were ...

Embodiment 2

[0059] Example 2 Detection of the effectiveness of the polypeptide having mucosal immune adjuvant activity of the present invention

[0060] 1. Experimental method

[0061] Different concentrations of the polypeptide with mucosal immune adjuvant activity prepared in Example 1 were combined with a certain concentration of EVP1 to detect the optimal concentration of the polypeptide to exert immune adjuvant activity.

[0062] The polypeptide having mucosal immune adjuvant activity and / or the EVP1 protein prepared in Example 1 was dissolved with PBS buffer. a) 24 nude mice (male) were purchased from the Animal Experiment Center of Chongqing Medical University, and randomly divided into 4 groups, 6 in each group.

[0063] b) The mucosal immune adjuvant polypeptide prepared in Example 1 was mixed with the EVP1 protein (10.0 μg / body) prepared in Example 2 according to the concentration of 5.0 μg / body in group A, 10.0 μg / body in group B, and 20.0 μg / body in group C. ) combination, a...

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Abstract

The invention discloses a polypeptide with mucosal immunity adjuvant activity, and application of the polypeptide in preparation of a mucosal immunity adjuvant. The polypeptide disclosed by the invention has the amino acid sequence as shown in SEQ ID NO:1. The polypeptide is a good mucosal immunity adjuvant, the mucosal immunity adjuvant activity of the polypeptide is equal to that of escherichia coli heat-labile enterotoxin LTB; meanwhile, the polypeptide is free of toxic and side effects, safe and effective, and can be combined with the vaccine antigen which is widely used at present, and can achieve a good clinical application prospect.

Description

technical field [0001] The invention relates to the fields of immunology and preventive medicine, in particular to a polypeptide with mucosal immune adjuvant activity and its use in preparing the mucosal immune adjuvant. Background technique [0002] The mucosal immune system (MIS) is an important part of the systemic immune system, including intestinal mucosa-associated lymphoid tissue, nasopharyngeal mucosa-associated lymphoid tissue, and urogenital tract mucosa-associated lymphoid tissue. Line of defense, MIS can effectively resist the invasion of various microorganisms. Compared with subcutaneous or intramuscular immunization, mucosal immunization has advantages over traditional immunization methods. The first is the difference in the way of immunization. Mucosal immunization mainly adopts the method of oral administration, nasal drip or genital tract inoculation. This new immunization method can avoid the stimulation of the body by the injection, reduce the inflammator...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K39/39A61P37/04
Inventor 马永平
Owner CHONGQING MEDICAL UNIVERSITY