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Vaccine to block transmission of echinococcosis pathogen Echinococcus granulosus at source

A technology of Echinococcus granulosus and vaccines, applied in the field of vaccines, can solve the problems of accelerating the control and elimination of echinococcosis, achieve the effects of increasing serum titer, improving immune level, and enhancing intestinal mucosal immune response

Active Publication Date: 2020-12-18
THE FIRST TEACHING HOSPITAL OF XINJIANG MEDICAL UNIVERCITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Scholars at home and abroad have made a lot of progress in this area. Anti-E.g vaccines have experienced dead vaccines, genetically engineered subunit protein vaccines, etc., but so far, there has been no vaccine for dogs and other terminal hosts. The development of tapeworm vaccine will greatly speed up the control and elimination of echinococcosis, which is an urgent task in the research of echinococcosis prevention and control

Method used

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  • Vaccine to block transmission of echinococcosis pathogen Echinococcus granulosus at source
  • Vaccine to block transmission of echinococcosis pathogen Echinococcus granulosus at source
  • Vaccine to block transmission of echinococcosis pathogen Echinococcus granulosus at source

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1, Echinococcus granulosus recombinant CTB-EgM123 vaccine and its immune effect detection

[0048] 1. Preparation of Echinococcus granulosus recombinant CTB-EgM123 vaccine

[0049] 1. Construction of recombinant prokaryotic expression vector

[0050] (1) Construction of recombinant prokaryotic expression vector pET28a / CTB-EgM123

[0051] First, according to the multi-cloning site on the prokaryotic expression vector pET28a, the CTB sequence was introduced into the pET28a vector, specifically: artificially synthesized CTB sequences with restriction sites Nhe I and BamH I at both ends (" GCTAGC +sequence 5+ GGATCC ”), and after digested by Nhe I and BamH I, it was connected to the large backbone fragment of the pET28a vector that had undergone the same digestion, and the intermediate vector pET28a / CTB was obtained after sequencing and verification.

[0052] Then, according to the EgM123 gene coding sequence published on GenBank and specific primers were desig...

Embodiment 2

[0091] Example 2. Detection of combined immune effect of Echinococcus granulosus recombinant CTB-EgM123 vaccine and Echinococcus granulosus EgM123 protein recombinant Mycobacterium smegmatis vaccine

[0092] 1. Preparation of Echinococcus granulosus EgM123 protein recombinant Mycobacterium smegmatis vaccine

[0093] 1. Construction of recombinant plasmid pMV261-EgM123

[0094] Primers were designed according to the published EgM123 sequence on GenBank, and the primer sequences were:

[0095] Primer-F2 (the underlined part is the recognition sequence of EcoR I):

[0096] 5'-CAT GAATTC ATGGAACCAGTGAATTTTGCC-3';

[0097] Primer-R2 (the underlined part is the recognition sequence of Sal I):

[0098] 5'-ACT GTC GAC TTATTTCGTCTTTAAGGCACGAAAC-3'.

[0099] Primers were synthesized by Shanghai Bioengineering Technology Service Co., Ltd. The EgM123 gene fragment was amplified using the adult cDNA of Echinococcus granulosus (the total RNA of the adult was extracted and obtained ...

Embodiment 3

[0112] Example 3, Echinococcus granulosus recombinant CTB-EgM123 vaccine enhances intestinal mucosal immune response

[0113] BALB / c mice were immunized with fusion protein CTB-EgM123 prepared and purified in Step 1 of Example 1, 20 μg / mouse for each immunization, and Freund's adjuvant was added at the same time. Freund's adjuvant (the first time is Freund's complete adjuvant, the second to third time is Freund's incomplete adjuvant) and the fusion protein CTB-EgM123 3 are completely mixed into an emulsion according to the ratio of 1:1 (volume ratio). Subcutaneous immunization. One week after the completion of the three immunizations, the small intestine was washed with normal saline, fixed with 4% paraformaldehyde, and routinely embedded in paraffin, sectioned, and stained with HE using the methods of tissue observation and immunohistochemical detection of the small intestine after immunization. Learn to observe. Goat anti mouse IgA(A90-403P), Goat anti mouse IgG2a(A90-107P...

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Abstract

The invention discloses a vaccine for blocking transmission of echinococcosis pathogeny echinococcus granulosus from a source. The vaccine provided by the invention is a CTB-EgM123 vaccine or a complete set of vaccine consisting of the CTB-EgM123 vaccine and a subunit vaccine of an EgM123 protein recombinant mycobacterium smegmatis vaccine. An active ingredient of the CTB-EgM123 vaccine is a fusion protein formed by fusion of a cholera toxin B subunit and an echinococcus granulosus EgM123 protein; the active ingredient of the EgM123 protein recombinant mycobacterium smegmatis vaccine can express the recombinant mycobacterium smegmatis of the echinococcus granulosus EgM123 protein. The vaccine provided by the invention can stimulate the immune response in a host, improves the immune level of the host, enhances the intestinal mucosal immune response, and plays an important protective role in resisting infection of echinococcosis.

Description

technical field [0001] The invention belongs to the field of vaccines, and relates to a vaccine for preventing and controlling the spread of Echinococcus granulosus, in particular to a vaccine for blocking the spread of echinococcus granulosus at the source. Background technique [0002] Echinococcosis is a serious zoonotic parasitic disease caused by Echinococcus larvae parasitizing in humans and some animals. More than 95% of echinococcosis cases are cystic echinococcosis (cystis echinococcosis, CE) caused by Echinococcus granulosus (E.g) infection, which is distributed worldwide. Nine provinces and regions in western my country (Xinjiang, Gansu, Ningxia, Qinghai, Sichuan, Inner Mongolia, Tibet, Yunnan and Heilongjiang provinces or autonomous regions) are high-endemic areas, and the prevalence rate of the population can be as high as 1% to 10%, seriously affecting agriculture and animal husbandry in western my country economic development and public health of the region. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/002A61K39/04A61P33/10
CPCA61K39/002A61K39/04A61K2039/70A61K2300/00
Inventor 张文宝李军张壮志郑雪婷何黎齐文静郭刚王俊伟郭宝平
Owner THE FIRST TEACHING HOSPITAL OF XINJIANG MEDICAL UNIVERCITY
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