A class of polypeptides that promote broad-spectrum immune responses in pigs and their applications

An immune response and pig body technology, applied in the field of biomedicine, can solve the problems of lack of immune enhancers, no data to prove broad-spectrum effect, complex components, etc., to promote broad-spectrum immune response, promote antigen-specific immune response, The effect of enhancing the immune response

Active Publication Date: 2021-12-03
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A class of polypeptides that promote broad-spectrum immune responses in pigs and their applications
  • A class of polypeptides that promote broad-spectrum immune responses in pigs and their applications
  • A class of polypeptides that promote broad-spectrum immune responses in pigs and their applications

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: MMIFLTYDL;

[0065] SEQ ID NO.2: LSIGYTLLF;

[0066] SEQ ID NO.3: HLNGFVSTL;

[0067] SEQ ID NO.4: ASGGGPAP;

[0068] SEQ ID NO.5: GIALDLSTL;

[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 1146g / 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 1026.6g / mol, which is consistent with the ...

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 inactivated ASFV strains (from the African Swine Fever Regional Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences) (10HID50). A booster immunization was given 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 tube, centrifuge at 1000r...

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 h. 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 subt...

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Abstract

The invention provides a class of polypeptides that can promote a broad-spectrum immune response in pig bodies and applications thereof, belonging to the technical field of biomedicine; the polypeptides include a first polypeptide, a second polypeptide, a third polypeptide, and a fourth polypeptide and one or more of the 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 as SEQ ID NO.1~ Shown in SEQ ID NO.5. The polypeptide of the present invention can significantly promote the activation of the IFN-β signaling pathway in the pig body, promote the secretion of IFN-γ by mononuclear macrophages, T cells and B cells, and promote the proliferation of spleen lymphocytes and mononuclear macrophages, thereby promoting The pig body produces a broad-spectrum immune response; moreover, the polypeptide of the invention can promote the ASFV antigen-specific immune response in the pig, so it can be used as an active ingredient of the ASFV subunit vaccine to enhance the immune response of the pig.

Description

technical field [0001] 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 [0002] 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. [0003] Immunopotentiators can stimulate the body to produce humoral-mediated and cell-mediated immune responses, so as to eliminate invading pathogens and protect animals from the harm of pathogens. [0004] There are many kinds of immune enhancers, which are generally divided into three categories according to their composition characteristics: Chinese herbal med...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06A61K39/39A61K39/12A61P37/04A61P31/20
CPCC07K7/06A61K39/39A61K39/12A61P37/04A61P31/20A61K2039/55516C12N2710/12034Y02A50/30
Inventor 郑海学毛箬青周晓丽孙德惠朱昱茜张克山张忠献杨帆刘华南王尧河刘湘涛
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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