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Chimeric HCV (hepatitis C virus) vaccine taking influenza virus as carrier and preparation method thereof

A technology of influenza virus and chimeric vaccine, applied in the field of HCV chimeric vaccine and its preparation, can solve the problems of weakened side effects, DNA vaccine safety, comprehensive response and limited protection

Active Publication Date: 2014-04-30
302 MILITARY HOSPITAL OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the journal "Science-Translational Medicine" in 2012, researchers from Oxford University and other institutions in the UK reported the results of a clinical trial of an HCV vaccine they developed, which uses a chimpanzee adenovirus as a vector to mimic its The internal structure is relatively stable for a long time, and clinical trials have achieved preliminary results. After 41 volunteers were vaccinated, follow-up studies found that the vaccine effect can last at least one year, and the side effects are significantly weakened, but the safety and comprehensive response and protection of DNA vaccines are affected. limit
And using influenza virus as the carrier of HCV vaccine development, there is no report at home and abroad

Method used

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  • Chimeric HCV (hepatitis C virus) vaccine taking influenza virus as carrier and preparation method thereof
  • Chimeric HCV (hepatitis C virus) vaccine taking influenza virus as carrier and preparation method thereof
  • Chimeric HCV (hepatitis C virus) vaccine taking influenza virus as carrier and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0064] Experimental example 1, the preparation of the HCV chimeric vaccine rFLU-HCV / NS1 of the chimeric HCV-C / E1 / E2 dominant epitope with influenza virus as the carrier

[0065] 1. The dominant epitope sequence of HCV-C / E1 / E2 in subtype 1b HCV is shown in SEQ ID No.1, which has been verified by animal experiments and serum of clinical patients, and meets the requirements of the next experiment.

[0066] 2. Construction of recombinant plasmid pC / E1 / E2-NS1

[0067] The recombinant plasmid pC / E1 / E2-NS1, the specific strategy is as follows figure 1 As shown in A.

[0068] Specifically, insert the HCV-C / E1 / E2 dominant epitope gene obtained above into the 5' end of the NS gene open reading frame (ORF) of the cold-adapted and attenuated influenza virus strain A / AA / 6 / 60 After the 375th nucleotide, a linker (5'-UAAUG-3') is added in the middle. The linker functions as both a terminator and a promoter, and finally the 376th from the 5' end of the open reading frame of the NS gene F...

experiment example 2

[0088] Experimental example 2, the preparation of the HCV chimeric vaccine rFLU-HCV / M of the chimeric HCV-C / E1 / E2 dominant epitope with influenza virus as the carrier

[0089] 1. The dominant epitope sequence of HCV-C / E1 / E2 in subtype 1b HCV is shown in SEQ ID No.1, which was verified by immunizing animals according to the method in step 1 of Example 1, and met the requirements of the next experiment.

[0090] 2. Construction of recombinant plasmid pC / E1 / E2-M

[0091] Using the M gene fragment of the cold-adapted and attenuated influenza virus strain A / AA / 6 / 60 as the target for inserting the dominant epitope gene of HCV-C / E1 / E2, the recombinant plasmid pC / E1 / E2-M, specific strategies such as figure 1 Shown in B.

[0092] Specifically, insert the HCV-C / E1 / E2 dominant epitope gene obtained above into the first 222 nucleosides of the open reading frame (ORF) of the M gene of the cold-adapted and attenuated influenza virus strain A / AA / 6 / 60 Between the acid and the last 222 nuc...

Embodiment 3

[0110] Embodiment 3, the preparation of the HCV chimeric vaccine rFLU-HCV / NA of the chimeric HCV-C / E1 / E2 dominant epitope with influenza virus as the carrier

[0111] 1. The dominant epitope sequence of HCV-C / E1 / E2 in subtype 1b HCV is shown in SEQ ID No.1, which was verified by immunizing animals according to the method in step 1 of Example 1, and met the requirements of the next experiment.

[0112] 2. Construction of recombinant plasmid pC / E1 / E2-NA

[0113] Using the NA gene fragment of A / California / 07 / 2009 (H1N1) as the target for inserting the dominant epitope gene of HCV-C / E1 / E2, the recombinant plasmid pC / E1 / E2-NA was constructed by molecular biology methods, specifically Strategies such as figure 1 C shown.

[0114] Specifically, the HCV-C / E1 / E2 dominant antigen epitope gene obtained above is inserted into the first 183 nucleosides of the NA gene open reading frame (ORF) of the influenza virus strain H1N1 subtype A / California / 07 / 2009 Between the acid and the last 15...

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Abstract

The invention discloses a chimeric HCV (hepatitis C virus) vaccine taking an influenza virus as a carrier and a preparation method thereof. The invention discloses DNA (deoxyribonucleic acid) molecules as shown in SEQ ID NO.1. The chimeric HCV vaccine disclosed by the invention is a bivalent vaccine, and a foundation is laid for achieving dual or multiple purposes of the chimeric HCV vaccine taking the influenza virus as the carrier.

Description

technical field [0001] The invention relates to an HCV chimeric vaccine with influenza virus as a carrier and a preparation method thereof. Background technique [0002] Hepatitis C is a deadly killer that endangers human health and life safety, and the prevention and control situation is particularly severe in developing countries. Hepatitis C virus (HCV) is the main pathogen that causes chronic liver disease, mainly in blood and mucous membranes in sexual life. Routes of infection transmission. According to the report of national authorities, there are about 170-200 million people infected with HCV in the world, and the rate of HCV infected people in my country is about 40 million or more. After HCV infection, about 50%-85% develop into chronic liver disease, of which 20%-30% develop into liver cirrhosis, and partly into hepatocellular carcinoma (HCC). At present, the combined application of polyethylene glycol, interferon-a and ribavirin is the standard regimen for the ...

Claims

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Application Information

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IPC IPC(8): C12N15/51C12N15/85C12N7/01A61K39/295A61K39/29A61K39/145A61P31/16A61P31/14C12R1/93
CPCA61K39/12A61K2039/5254A61K2039/5256A61P31/14A61P31/16C07K14/005C12N2760/16034C12N2760/16043C12N2770/24034C12N2770/24234C12N2799/021
Inventor 杨鹏辉张绍庚王希良段跃强张培瑞李志伟王兆海
Owner 302 MILITARY HOSPITAL OF CHINA
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