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Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof

An immune response, pig body technology, applied in the field of biomedicine, can solve the problems of complex components, lack of immune enhancers, and high production costs, and achieve the effect of enhancing immune response and promoting antigen-specific immune response.

Active Publication Date: 2022-01-14
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that CVC1320 is used as an immune enhancer for foot-and-mouth disease vaccines (Studies on how immune enhancers improve the immune efficacy of porcine foot-and-mouth disease inactivated vaccines, Yu Xiaoming, 2019), but the immune enhancer is a compound preparation with complex components and large dosages. relatively high production costs
Moreover, the preparation is only verified to be applicable to FMDV vaccine, and there is no data to prove that it has a broad-spectrum effect
Tang Bo et al. (Studies on Immunopotentiators Improving the Immune Efficacy of Inactivated Vaccines for Pigs, 2016) used VA5-containing immunopotentiators as companions of porcine parvovirus and JE dual inactivated vaccines, which could increase the antibody titer, but there was no data to prove its efficacy. have a broad-spectrum effect
Currently, there is an urgent need for an immune enhancer that induces a broad-spectrum immune response in pigs

Method used

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  • Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof
  • Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof
  • Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Solid phase synthesis and purity detection of embodiment 1 polypeptide

[0063] The polypeptide of the present invention is as follows:

[0064] SEQ ID NO.1: KMRIEIFWEL;

[0065] SEQ ID NO.2: RSMMMVFNQL;

[0066] SEQ ID NO.3: YGIALQYVL;

[0067] SEQ ID NO.4: IFNFIENIWS;

[0068] SEQ ID NO.5: YTEMLALII;

[0069] The above polypeptides were synthesized by Shanghai Sangon Bioengineering Co., Ltd.

[0070] The detection wavelength is 214nm. The purity of the final peptide purification product was >98%, and the structure was identified by ESI-MS. For the identification results, see Figure 1 to Figure 10 ,in figure 1 is the identification chromatogram of AP1, figure 2 It is the mass spectrum of AP1, and the identified molecular weight is 1364.85g / mol, which is consistent with the theoretical value; image 3 is the identification chromatogram of AP2, Figure 4 It is the mass spectrum of AP2, and the identified molecular weight is 1256.6g / mol, which is consistent wi...

Embodiment 2

[0071] Embodiment 2 porcine lymphocyte proliferation experiment

[0072] 1. Pigs immunized with inactivated ASFV virus.

[0073] Five 90-day-old male Landrace pigs were immunized with ASFV epidemic strain inactivated virus (from the African swine fever regional laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences) (10HID50), and a booster immunization was performed one month later, and the pigs were administered 7 days later. Euthanize and remove the spleen after dissection. Unimmunized healthy pigs were used as negative control group.

[0074] 2. Preparation, culture and proliferation detection of splenocytes.

[0075] 1) Aseptically treat the collected pig spleen with 75% alcohol, wash it three times with PBS, cut the spleen into small pieces, place them in folded sterile gauze (2 layers), and place them in 5 mL of serum-containing 1640 medium Grind the spleen in a plate.

[0076] 2) Then draw the liquid into a 15ml centrifuge tub...

Embodiment 3

[0085] Example 3 Detection of IFN-γ secretion by different subtypes of splenic lymphocytes.

[0086] The pig immunization procedure and spleen cell isolation and culture are the same as in Example 2. After obtaining the dispersed splenocytes, prepare a single cell suspension with RPMI1640 complete medium at a concentration of 1×10 6 / ml. Inoculate into a 24-well plate, add 0.2 μg of the polypeptide provided by the present invention to each well, and culture in a CO2 incubator for 60 hours. Collect the cells in each well, label the cells with porcine CD3, CD4, CD8 and IFN-γ specific antibodies respectively, then wash twice with PBS buffer containing 2% serum, and finally use the washing solution to disperse into a cell suspension, and use the flow Cytometer detection, CD4 respectively + T lymphocytes and CD8 + Levels of IFN-γ secreted by T lymphocytes.

[0087] For test results, see Figure 13 ~ Figure 17 . in Figure 13 Promote the level of IFN-γ secreted by different ...

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Abstract

The invention provides a polypeptide for promoting a pig body to generate broad-spectrum immune response and application thereof, and belongs to the technical field of biomedicine. The polypeptide comprises one or more of a first polypeptide, a second polypeptide, a third polypeptide, a fourth polypeptide and a fifth polypeptide; the amino acid sequences of the first polypeptide, the second polypeptide, the third polypeptide, the fourth polypeptide and the fifth polypeptide are respectively shown as SEQ ID NO. 1 to SEQ ID NO. 5. The polypeptide provided by the invention can promote the pig body to generate broad-spectrum immune response by simultaneously promoting the proliferation of lymphocytes and mononuclear macrophages in the pig body; moreover, the polypeptide provided by the invention can promote the specific immune response of ASFV antigen in the pig body, so that the polypeptide can be used as an effective component of an ASFV subunit vaccine to enhance the immune response of pigs.

Description

[0001] This application is a divisional application with an application date of August 26, 2020, an application number of CN202010871941.8, and an invention title of "A Class of Polypeptides and Applications for Promoting Broad-spectrum Immune Response in Pigs". technical field [0002] The invention relates to the technical field of biomedicine, in particular to a class of polypeptides for promoting a broad-spectrum immune response in pig bodies and applications thereof. Background technique [0003] Immunopotentiators are a class of agents that are used alone or in combination with antigens to enhance the immune system of animals by enhancing the activity of macrophages, enhancing the immunogenicity and stability of antigenic substances, and promoting the synthesis and secretion of various immune factors and specific antibodies. Substances of the body's immune response. [0004] Immunopotentiators can stimulate the body to produce humoral-mediated and cell-mediated immune ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K39/39A61K39/12A61P37/04A61P31/20
CPCC07K7/06A61K39/39A61K39/12A61P37/04A61P31/20A61K2039/55516C12N2710/12034
Inventor 郑海学杨帆毛箬青周晓丽孙德惠朱昱茜张克山刘华南刘湘涛
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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