Porcine circovirus, porcine parvovirus and porcine reproductive and respiratory syndrome virus triple virus-like particle vaccine and its preparation method

A technology for respiratory syndrome and porcine circovirus, applied in antiviral agents, viral antigen components, pharmaceutical formulations, etc., can solve the problems of long preparation period, no potential danger, no safety, etc.

Inactive Publication Date: 2012-06-13
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, compared with live vaccines and inactivated vaccines, VLP vaccines have no unsafe factors; compared with subunit vaccines, VLPs are composed of multiple antigenic proteins and have strong immunogenicity; Compared with Virosome, VLP is neither prepared from live virus, has no potential danger, does not require a large number of reproductive viruses, and has no disadvantages such as low safety, high cost, and long preparation cycle; thus VLP vaccines are widely used. Today is hailed as a new milestone in the development of viral vaccines

Method used

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Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0029] Experimental example 1: Triple VLP vaccine verification experiment of the present invention

[0030] The Triple VLP vaccine injection, nasal drops, and drinking water formulations prepared in Example 8 of the present invention are uniform in appearance, without peculiar smell and bacterial contamination; they have no pyrogen, abnormal toxicity and acute toxicity through guinea pig and rabbit tests; Intracerebral inoculation in rats and intramuscular injection in the neck of 3-day-old piglets showed no symptoms of disease; the protein content determined by the Lorry method was within the dose range; the diameter of VLP was about 17nm observed by electron microscopy, which was consistent with natural viruses; the DNA content determined by ELISA was within the range Below 50EU; detect the presence of antigenic protein components by SDS-PAGE and WB; immunize Balb / C mice once and the serum neutralizing antibody titer is above 3200.

experiment example 2

[0031] Experimental Example 2: Experiment of the immune response effect of Triple VLP vaccine of the present invention on experimental pigs

[0032] The Triple VLP vaccine prepared in Example 8 of the present invention was prepared by adding or not adding corresponding adjuvants to the injection, nasal drop, and drinking water vaccine dosage forms, and used PBS, aluminum salt, nano-aluminum, rLT as the control group, and immunized for 3 days according to their respective routes. age piglets, and its immunization dosage of back-up sows is immunized twice (0, 21 days) by embodiment 8 vaccine low-dose groups; Collect the peripheral blood of each group piglets and sows, separated serum and For immune cells, collect splenic lymphocytes, lung and nasal lavage fluid; measure the serum antibody titer above 6400 by ELISA method, and measure the neutralizing antibody titer above 3200 by MTT method; use flow cytometer and ELISPOT instrument to measure immune cells Dynamic changes in seru...

experiment example 3

[0033] Experimental Example 3: Experiment of the immune protection effect of Triple VLP vaccine of the present invention on experimental pigs

[0034] The Triple VLP vaccine prepared in Example 8 of the present invention added or not added the corresponding adjuvant prepared injection, nasal drops, drinking water formulations as the experimental group, and with PBS, aluminum salt, nano-aluminum, rLT, and PCV-2, PPV, PRRSV formaldehyde The inactivated virus was a control group, and 3 day-old piglets and gilts were immunized respectively by their respective approaches. The immunization dose was immunized twice (0, 21 days) by the vaccine low-dose group in Example 8, and each time was 14 days after the last immunization. Experimental group, control group experimental pigs use 10 5.0 TCID 50Clinically isolated PCV-2, PPV, and PRRSV strains were challenged by intramuscular injection in the neck to observe the protective effect. The result shows that Triple VLP vaccine of the pres...

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Abstract

The purpose of the invention is to disclose a porcine circovirus, porcine parvovirus and porcine reproductive and respiratory syndrome virus triplex virus-like particle vaccine and its preparation method. The triple virus-like particle vaccine (Triple VLP vaccine) of the invention contains VLP which is composed of PCV-2 major structural protein CAP protein, PPV VP2 protein epitope and PRRSV Gp5 protein epitope. It is proved by experiment that the vaccine can stimulate good double cellular and humoral immune response. It is shown by pharmacodynamic test that after immunization of different animal groups, the vaccine of injection, nose drops and water forms prepared by VLP antigen formed by the method with or without adjuvants can safely and effectively prevent the infection of PCV-2, PPV and PRRSV. The invention provides an ideal vaccine for the security of sows, piglets and fattening pigs to effectively prevent mixed infection of PCV-2, PPV and PRRSV.

Description

field of invention [0001] The invention belongs to the field of biopharmaceuticals, and relates to a porcine circovirus, porcine parvovirus, porcine reproductive and respiratory syndrome virus triple virus-like particle vaccine and a preparation method thereof, in particular to a type-2 porcine circovirus-containing nuclear vaccine A triple virus-like particle vaccine (Trivalent VLP) composed of capsid protein, porcine parvovirus VP2 protein epitope, and porcine reproductive and respiratory syndrome virus Gp5 protein and a preparation method thereof. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) will lead to decreased immunity of pig herds, and is associated with postweaning multisystemic wasting syndrome (Postweaning Multisystemic Wasting Syndrome, PMWS), porcine dermatitis and nephrotic syndrome (Porcine Dermatitis and Nephropathy Syndrome, PDNS), Porcine reproductive failure, Porcine Respiratory Disease Syndrome (PRDC), Porcine Necrotizing Pne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295A61K48/00C12N15/79A61P31/14A61P31/20A61K39/12A61K39/23
Inventor 吕凤林潘玉竹曹政
Owner CHONGQING UNIV
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