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CpG-ODN with specific immunostimulation effect on PRRSV and application of CpG-ODN

An immunostimulatory and specific technology, applied in the field of CpG-ODN, to achieve the effect of improving the induction of immune response, reducing the cost of pig breeding and reducing the incidence of disease

Active Publication Date: 2021-02-05
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the immunostimulatory activity of two immune activation units (-CG-motif and -GTCGTT-motif) at the same time.

Method used

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  • CpG-ODN with specific immunostimulation effect on PRRSV and application of CpG-ODN
  • CpG-ODN with specific immunostimulation effect on PRRSV and application of CpG-ODN
  • CpG-ODN with specific immunostimulation effect on PRRSV and application of CpG-ODN

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Experimental program
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Effect test

Embodiment

[0028] 1. Screening of specific CpG-ODN fragments

[0029] Screening method: According to conventional techniques, CpG-ODN fragments were directional screened, and the results are shown in Table 1 below.

[0030] Table 1 Screened CpG-ODN fragments

[0031]

[0032] The CpG-ODN fragments screened above are screened based on the research of related sequences by relevant personnel, for example:

[0033] References for C274 are "Marshall, J.D., Higgins, D., Abbate, C., et al. (2004). Polymyxin B enhances ISS-mediated immune responses across multiple species. Cellular Immunology, 229, 93–105." and "Teleshova , N., Kenney, J., Williams, V., et al. (2006). CpG-C ISS-ODN activation of blood-derived B cells from healthy and chronic immunodeficiency virus-infected macaques. Journal of Leukocyte Biology, 79, 257–267 "."

[0034]BCG-A4a reference is "Yamamoto, S., Yamamoto, T., Nojima, Y., et al. (2002). Discovery of immunostimulatory CpG-DNA and its application to tuberculosis vacc...

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Abstract

The invention relates to a CpG-ODN with a specific immunostimulation effect on PRRSV and an application of the CpG-ODN. 11 CpG-ODN fragments are screened out and then connected end to end to form a DNA sequence containing a plurality of -CG-dinucleotide as a CpG element, and the DNA sequence is named as "Multi-CpG". A section of the DNA sequence is replaced with the same length in the "Multi-CpG"with the DNA sequence taking -GTCGTT- as the CpG element, and the DNA sequence is named as "Multi-CpG-Mix". Two "Multi-CpG" and one "Multi-CpG-Mix" are connected end to end to obtain the DNA sequencecontaining two immune activation units, and the DNA sequence is named as "Multi-CpG-Mixture". The "Multi-CpG-Mixture" is inserted into a multi-copy clone vector Puc19 derived vector FVX through two restriction enzyme cutting sites, i.e., XbaI and EcoRI, and a recombinant plasmid FVX-Multi-CpG-Mixture is constructed by virtue of a gene synthesis means. The CpG-ODN is capable of inducing natural immune function activation in a pig body, activating acquired immune function of the PRRSV in the pig body, improving the immune response induction effect of the PRRSV in the pig body, reducing the PRRSVmorbidity and reducing the pig breeding cost.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a CpG-ODN with specific immunostimulatory effect on PRRSV and its application. Background technique [0002] Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the causative agent of Porcine Reproductive and Respiratory Syndrome (PRRS). PRRSV was first discovered in the United States in 1987. In 1991, the Dutchman Wensvoot isolated the virus from sick piglets and sows for the first time, and then Germany, the United States, and the United Kingdom also isolated the virus. At present, the virus has spread throughout North America and Europe, and has spread globally, and epidemic infections have occurred successively in various places, causing serious economic losses to the pig industry. Although porcine reproductive and respiratory syndrome virus vaccines have been widely used, they are accompanied by side effects and have only partial immune protection, which m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/117C12N15/85A61K48/00A61P31/14
CPCA61K48/005A61P31/14C12N15/117C12N15/85C12N2310/17C12N2320/30C12N2800/107
Inventor 冯璥李涛何文彬吕雪梅
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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