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PRRSV attenuated strain which is capable of inducing pig body to relatively early generate interferon and neutralizing antibody and possesses wide-spectrum immunogenicity

An immunogenic and interferon technology, applied in the field of medicine and biology, can solve the problems of unsatisfactory effects of attenuated live vaccines and inactivated vaccines, and poor immune effects of heterologous genotype strain infection, so as to improve immunosuppression And delayed immune response, reduce viral infection, long-lasting effect of immune protection

Active Publication Date: 2015-03-04
张澍 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Porcine reproductive and respiratory syndrome virus escapes the body's immunity and virus diversity has led to unsatisfactory effects of attenuated live vaccines and inactivated vaccines currently on the market at home and abroad, especially for heterologous genotype strains.
Experimental porcine reproductive and respiratory syndrome virus vaccines, live attenuated vaccines, recombinant vector-expressed PRRSV protein vaccines, DNA vaccines, and plant-produced subunit vaccines developed by reverse molecular genetics have all been tried on different experimental animals, but due to The genetic and antigenic diversity of the virus itself, as well as the poor understanding of its pathogenicity and immunity, constitute a major challenge for the development of these experimental vaccines

Method used

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  • PRRSV attenuated strain which is capable of inducing pig body to relatively early generate interferon and neutralizing antibody and possesses wide-spectrum immunogenicity
  • PRRSV attenuated strain which is capable of inducing pig body to relatively early generate interferon and neutralizing antibody and possesses wide-spectrum immunogenicity
  • PRRSV attenuated strain which is capable of inducing pig body to relatively early generate interferon and neutralizing antibody and possesses wide-spectrum immunogenicity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Porcine Reproductive and Respiratory Syndrome Virus Isolation, Passaging and Related Identification of Pathogenicity and Immunogenicity

[0028] 1. Materials and methods

[0029] 1.1 Material

[0030] Piglets are 3 weeks old, no specific pathogens and no PRRSV antibodies, commercial PRRSV antibody detection ELISA kit (IDEXX Laboratories, Inc., Westbrook, Mass.), test strains ATCC VR2385, ATCC VR2332 purchased from the United States ATCC, HP-PRRSV- HuN4-F112 (lot JA-621C-416) is purchased from the market, 2ml is 1 serving.

[0031] 1.2 method

[0032] 1.2.1 Virus isolation and passage

[0033] A typical and severe porcine reproductive and respiratory syndrome broke out in a pig farm in Baoding, Hebei. 80 of the 300 sows in the pig farm had aborted within 5 weeks, 50% were weaned early, 15% of fattening pigs died, and pig mortality 78%. Blood was collected from sick pigs, the serum was filtered with a 0.45um filter, and the filtrate was placed at -60°C for use. 75cm 2 ...

Embodiment 2

[0063] Example 2 Study on the stability (pathogenicity and attenuation) of the original seed batch PRRSV-LZ-F65 in pigs

[0064] 1. Method:

[0065] Pigs were inoculated with PRRSV-LZ-F65. Pigs were divided into 6 groups. In the first group, 5 pigs were inoculated intranasally with PRRSV-LZ-F65, and 3 pigs in the control group were inoculated intranasally with sterile liquid. The clinical symptoms: appearance, breathing, Stool etc. Weighed, bled, and slaughtered after 6 days. Collect the lung lavage material of each pig body and freeze-thaw it; collect the pig serum to prepare a mixture of 2ml of each pig lung lavage material and 2ml serum for inoculation of the pigs of the second group and the second control group. The inoculation was successively passaged to the sixth generation, and the observation and collection of lung lavage and serum were repeated in each generation. Each group is raised separately but under the same conditions.

[0066] After the pigs were inoculated with...

Embodiment 3

[0077] Example 3 Porcine Reproductive and Respiratory Syndrome Virus Attenuated Strain HP-PRRSV-LZ-F65 Seed Batch Virus and the Immunogenicity and Safety Experiments of the Passage Virus

[0078] 1. Material

[0079] 1.1 Experimental animals: 100-120 4-week-old pigs are negative for antibodies to PRRSV and Mycoplasma pneumonia, and PCR test for porcine circovirus type 2 is negative. Divide into groups A-F and raise them in isolation.

[0080] Test vaccine: Porcine Reproductive and Respiratory Syndrome Vaccine HP-PRRSV-LZ-F95, HP-PRRSV-LZ-F80, HP-PRRSV-LZ-110 (HP-PRRSV-LZ-F65 is the original seed batch of the vaccine, and the 80 generation is the main Seed batch, 110 generation is the working seed batch, using MARC-145 cell subculture, the specific method is as in Example 1), domestic porcine reproductive and respiratory syndrome vaccine HP-PRRSV-HuN4-F112, foreign imported vaccine Ingelvac PRRS MLV (Boehringer Ingelheim, USA) purchased from the market. The porcine reproductive and...

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Abstract

The invention discloses a PRRSV attenuated strain which is capable of inducing pig body to relatively early generate interferon and a neutralizing antibody and possesses wide-spectrum immunogenicity. The porcine reproductive and respiratory syndrome virus (PRRSV) attenuated strain obtained through separation, passage and attenuation is named as HP-PRRSV-LZ-F65, and is prepared by subculturing PRRSV isolated virus in primary porcine alveolar macrophage for 5 generations and in MARC-145 cell for 60 generations and then performing attenuation, and the homologous rates of HP-PRRSV-LZ-F65 with PRRSV VR-2332 and VR-2385 are respectively 97.50% and 98.2%. Experiments prove that PRRSV-LZ-F65 is capable of relatively early inducing a pig body to generate an anti-PRRSV neutralizing antibody with the level substantially higher than that of other same-kind vaccines, and also is capable of preventing challenge infection of homologous type and heterogenous type PRRSV, and possesses stable gene after being subcultured in vitro for 50 generations. The attenuated strain HP-PRRSV-LZ-F65 and passage strains thereof possess wide application prospect to prevent porcine reproductive and respiratory syndrome as safe efficient wide-spectrum vaccine strain candidates.

Description

Technical field [0001] The invention relates to an attenuated strain of porcine reproductive and respiratory syndrome virus and a vaccine thereof, in particular to a porcine reproductive and respiratory syndrome virus that can induce early production of interferon and neutralizing antibodies in pigs and has broad-spectrum immunogenicity An attenuated strain and a vaccine prepared from the attenuated strain or its passage strains. The invention belongs to the field of medical biotechnology. Background technique [0002] Porcine reproductive and respiratory syndrome virus (PRRSV), a member of the Arteriviridae Arteritis virus genus of the Arteritis virus family of the order Necrovirus, is the pathogen that causes porcine reproductive and respiratory syndrome and can cause pregnant sows Late miscarriage, premature delivery and stillbirth, respiratory diseases in piglets and fattening pigs. [0003] The genome of porcine reproductive and respiratory syndrome virus is a single-strande...

Claims

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Application Information

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IPC IPC(8): C12N7/08A61K35/765A61K39/12A61P31/14C12R1/93
Inventor 张澍马忠仁冯若飞吕宏亮
Owner 张澍
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