CN113929746B - Polypeptide for promoting pig organism to generate broad-spectrum immune response and application thereof - Google Patents
Polypeptide for promoting pig organism to generate broad-spectrum immune response and application thereof Download PDFInfo
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- CN113929746B CN113929746B CN202111306043.9A CN202111306043A CN113929746B CN 113929746 B CN113929746 B CN 113929746B CN 202111306043 A CN202111306043 A CN 202111306043A CN 113929746 B CN113929746 B CN 113929746B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55516—Proteins; Peptides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/12011—Asfarviridae
- C12N2710/12034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Abstract
The invention provides a polypeptide for promoting a pig organism to generate broad-spectrum immune response and application thereof, belonging 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-SEQ ID NO. 5. The polypeptide can promote the pig organism to generate broad-spectrum immune response by promoting the proliferation of lymphocytes and mononuclear macrophages in the pig organism; in addition, the polypeptide of the invention can promote the antigen specific immune response of the ASFV in pigs, thus being used as the effective component of the ASFV subunit vaccine and enhancing the immune response of pigs.
Description
The application is a divisional application of 26 days of the application date 2020 and the application number CN202010871941.8, and the invention is named as a polypeptide for promoting the pig organism to generate broad-spectrum immune response and application thereof.
Technical Field
The invention relates to the technical field of biomedicine, in particular to a polypeptide for promoting a pig organism to generate broad-spectrum immune response and application thereof.
Background
The immunopotentiator is a substance which can be used singly or in combination with antigen to enhance the immune response of animal organism by enhancing macrophage activity, enhancing the immunogenicity and stability of antigen substances, promoting the synthesis and secretion of various immune factors and specific antibodies, and the like.
The immunopotentiator can stimulate the organism to generate humoral-mediated and cell-mediated immune response, thereby eliminating the invading pathogen and protecting animals from the pathogen.
Immunopotentiators are of various kinds and are largely classified into three main categories according to their component characteristics: chinese herbal extracts, chemical compositions and cytokines. There are few immunopotentiators currently used in swine vaccines. CVC1320 is reported to be used as an immunopotentiator of a foot-and-mouth disease vaccine (research on improving the immunopotentiator for improving the immunopotentiator of a swine foot-and-mouth disease inactivated vaccine, yu Xiaoming and 2019), but the immunopotentiator is a compound preparation, has complex components, large using dosage and relatively high production cost. Moreover, the preparation is only verified to be applicable to FMDV vaccines, and no data prove that the preparation has a broad-spectrum effect. Tang Bo et al (study of immunopotentiator improving the immunopotentiator of inactivated vaccine for pig, 2016) used VA 5-containing immunopotentiator as a dual inactivated vaccine partner for pig's tiny, japanese encephalitis, and could improve antibody titer, but no data demonstrated broad-spectrum effect. Currently, there is a lack of an immunopotentiator that promotes a broad spectrum immune response in the swine organism.
Disclosure of Invention
The invention aims to provide a polypeptide for promoting pig organism to generate broad-spectrum immune response and application thereof, wherein the polypeptide can promote pig organism to generate broad-spectrum immune response by promoting mononucleated macrophages, T cells and B cells in pig organism to secrete IFN-gamma, and can be used as an active ingredient of ASFV subunit vaccine to enhance pig immune response.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a polypeptide for promoting a pig organism to generate broad-spectrum immune response; 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 sequence of the first polypeptide is shown as SEQ ID NO. 1;
the amino acid sequence of the second polypeptide is shown as SEQ ID NO. 2;
the amino acid sequence of the third polypeptide is shown as SEQ ID NO. 3;
the amino acid sequence of the fourth polypeptide is shown as SEQ ID NO. 4;
the amino acid sequence of the fifth polypeptide is shown as SEQ ID NO. 5;
the polypeptide promotes the pig body to generate broad-spectrum immune response by promoting the proliferation of lymphocytes and mononuclear macrophages in the pig body.
Preferably, the lymphocytes include spleen lymphocytes, peripheral blood T cells, and B cells in peripheral blood.
Preferably, the mononuclear macrophages include spleen mononuclear macrophages and peripheral blood mononuclear macrophages.
The invention provides a polypeptide polymer obtained by polymerizing the polypeptide in the scheme.
The invention provides application of the polypeptide or the polypeptide polymer in the scheme in preparation of a preparation for promoting a pig organism to generate a broad-spectrum immune response.
The invention provides application of the polypeptide or the polypeptide polymer in the scheme in preparation of a preparation for promoting swine organisms to generate african swine fever virus antigen specific immune response.
The invention provides a pig immunopotentiator, and the active ingredients of the immunopotentiator comprise the polypeptide or the polypeptide polymer in the scheme.
Preferably, the immunopotentiator has a content of the polypeptide or the polypeptide polymer of 100. Mu.g/component/head.
The invention provides a subunit vaccine of African swine fever virus, which comprises the polypeptide or the polypeptide polymer in the scheme.
Preferably, the content of the polypeptide or the polypeptide polymer in the subunit vaccine is 100 mug/component/head.
The invention has the beneficial effects that: the invention provides a polypeptide for promoting a pig organism to generate broad-spectrum immune response; 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-SEQ ID NO. 5. The polypeptide can promote the pig organism to generate broad-spectrum immune response by promoting the proliferation of lymphocytes and mononuclear macrophages in the pig organism; in addition, the polypeptide of the invention can promote the antigen specific immune response of the ASFV in pigs, thus being used as the effective component of the ASFV subunit vaccine and enhancing the immune response of pigs.
Drawings
FIG. 1 is an identification chromatogram of AP 1;
FIG. 2 is a mass spectrum of AP 1;
FIG. 3 is an identification chromatogram of AP 2;
FIG. 4 is a mass spectrum of AP 2;
FIG. 5 is an identification chromatogram of AP 3;
FIG. 6 is a mass spectrum of AP 3;
FIG. 7 is an identification chromatogram of AP 4;
FIG. 8 is a mass spectrum of AP 4;
FIG. 9 is an identification chromatogram of AP 5;
FIG. 10 is a mass spectrum of AP 5;
FIG. 11 is a flow chart of data statistics for the proliferation of AP 1-AP 5-activated lymphocytes and monocytes;
FIG. 12 is a flow chart of statistics of AP 1-AP 5 promoting proliferation of healthy porcine lymphocytes and mononuclear macrophages;
FIG. 13 is a graph showing that AP1 promotes IFN-gamma secretion by immune cells of different subtypes;
FIG. 14 is a graph showing that AP2 promotes IFN-gamma secretion by various subtypes of immune cells;
FIG. 15 shows the levels of AP3 promoting secretion of IFN-gamma by various subtypes of immune cells;
FIG. 16 shows that AP4 promotes IFN-gamma secretion by various subtypes of immune cells;
FIG. 17 is a graph showing that AP5 promotes IFN-gamma secretion by various subtypes of immune cells;
FIG. 18 shows the proportion of the level of the sub-population of B lymphocytes in the secondary claim 14 d;
FIG. 19 is CD8 + T lymphocyte subpopulation proportion level;
FIG. 20 shows the proportion of the level of the sub-population of B lymphocytes in the secondary claim 14 d;
FIG. 21 is CD8 + T lymphocyte subpopulation proportion level.
Detailed Description
The invention provides a polypeptide for promoting a pig organism to generate broad-spectrum immune response; the polypeptide comprises one or more of a first polypeptide, a second polypeptide, a third polypeptide, a fourth polypeptide and a fifth polypeptide; the polypeptide promotes the pig body to generate broad-spectrum immune response by promoting the proliferation of lymphocytes and mononuclear macrophages in the pig body simultaneously; the lymphocytes preferably include spleen lymphocytes, peripheral blood T cells and B cells in peripheral blood; the mononuclear macrophages preferably include spleen mononuclear macrophages and peripheral blood mononuclear macrophages.
In the invention, the amino acid sequence of the first polypeptide is shown as SEQ ID NO.1, and specifically comprises the following steps: KMRIEIFWEL (Lys-Met-Arg-Ile-Glu-Ile-Phe-Trp-Glu-Leu) has an average molecular weight of 1364.65g/mol and a chemical formula: c (C) 65 H 101 N 15 O 15 S, S. The theoretical isoelectric point of the polypeptide is pH7, the GRAVY value of the polypeptide is 0.12, and the polypeptide has hydrophobicity. This property facilitates binding to host proteins or cell surfaces and is more effectiveAn immune response is initiated.
In the invention, the amino acid sequence of the second polypeptide is shown as SEQ ID NO.2, and specifically comprises the following steps: RSMMMVFNQL (Arg-Ser-Met-Met-Met-Val-Phe-Asn-Gln-Leu) and has an average molecular weight of 1256.56g/mol and a chemical formula: c (C) 53 H 89 N 15 O 14 S 3 . The polypeptide has a theoretical isoelectric point of pH 11.05, a GRAVY value of 0.42, hydrophobicity, and can be combined with host protein or cell surface to effectively start immune response.
In the invention, the amino acid sequence of the third polypeptide is shown as SEQ ID NO.3, and specifically comprises the following steps: YGIALQYVL (Tyr-Gly-Ile-Ala-Leu-Gln-Tyr-Val-Leu) has an average molecular weight of 1039.22g/mol and a chemical formula: c (C) 51 H 78 N 10 O 13 . The polypeptide has a theoretical isoelectric point of pH7, a GRAVY value of 1.29, hydrophobicity, and can be combined with host protein or cell surface to effectively start immune response.
In the invention, the amino acid sequence of the fourth polypeptide is shown as SEQ ID NO.5, and specifically comprises the following steps: IFNFIENIWS (Ile-Phe-Asn-Phe-Ile-Glu-Asn-Ile-Trp-Ser) has an average molecular weight of 1282.44g/mol and a chemical formula: c (C) 63 H 87 N 13 O 16 . The theoretical isoelectric point of the polypeptide is pH 3.28, the GRAVY value of the polypeptide is 0.69, and the polypeptide has hydrophobicity.
In the invention, the amino acid sequence of the fifth polypeptide is shown as SEQ ID NO.4, and specifically comprises the following steps: YTEMLALII (Tyr-Thr-Glu-Met-Leu-Ala-Leu-Ile-Ile) has an average molecular weight of 1066.31g/mol and a chemical formula: c (C) 50 H 83 N 9 O 12 S, S. The polypeptide has a theoretical isoelectric point of pH 3.28, a GRAVY value of 1.64, hydrophobicity, and can be combined with host protein or cell surface to effectively start immune response.
In the invention, the polypeptide in the technical scheme is obtained by determining the sequence of an ASFV epidemic strain and by computer-aided bioinformatics prediction, and has specificity.
In the present invention, the polypeptide is preferably synthesized by Shanghai Biotechnology Co., ltd.
The present invention provides a polypeptide polymer comprising the polypeptide of the above scheme.
The invention provides application of the polypeptide or the polypeptide polymer in the scheme in preparation of a preparation for promoting a pig organism to generate a broad-spectrum immune response.
In the present invention, the polypeptide or the polypeptide polymer promotes the generation of a broad spectrum immune response in the pig body by promoting proliferation of lymphocytes and monocytes and macrophages in the pig body at the same time.
The invention provides application of the polypeptide or the polypeptide polymer in the scheme in preparation of a preparation for promoting swine organisms to generate african swine fever virus antigen specific immune response; the polypeptide or the polypeptide polymer can promote the pig organism to generate the African swine fever virus antigen specific immune response by promoting the proliferation of spleen lymphocytes and mononuclear macrophages of the pig organism.
The invention provides a pig immunopotentiator, the active ingredient of which comprises the polypeptide or the polypeptide polymer in the scheme; the content of the polypeptide or the polypeptide polymer in the immunopotentiator is preferably 100. Mu.g/component/head, i.e. the content of each polypeptide or each polypeptide polymer is 100. Mu.g/head.
The invention provides a subunit vaccine of African swine fever virus, which comprises the polypeptide or the polypeptide polymer in the scheme; the content of the polypeptide or the polypeptide polymer in the subunit vaccine is preferably 100 μg/component/head, i.e. the content of each polypeptide or each polypeptide polymer is 100 μg/head.
The formulations of the above-described regimens of the invention or the immunopotentiators of the above-described regimens or the subunit vaccine of the above-described regimens, respectively, comprise injections.
The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1 solid phase Synthesis and purity detection of Polypeptides
The polypeptides of the invention are as follows:
SEQ ID NO.1：KMRIEIFWEL；
SEQ ID NO.2：RSMMMVFNQL；
SEQ ID NO.3：YGIALQYVL；
SEQ ID NO.4：IFNFIENIWS；
SEQ ID NO.5：YTEMLALII；
the above polypeptides were synthesized by Shanghai Biotechnology Co., ltd.
The detection wavelength was 214nm. The purity of the final polypeptide purified product is more than 98%, and the structure is identified by ESI-MS, wherein the identification result is shown in figures 1-10, the figure 1 is an identification chromatogram of AP1, the figure 2 is a mass spectrum of AP1, and the identification molecular weight is 1364.85g/mol and is consistent with the theoretical value; FIG. 3 shows an identification chromatogram of AP2, FIG. 4 shows a mass spectrum of AP2, and the identification molecular weight is 1256.6g/mol, which is consistent with the theoretical value; FIG. 5 shows an identification chromatogram of AP3, FIG. 6 shows a mass spectrum of AP3, and the identification molecular weight is 1039.6g/mol, which is consistent with the theoretical value; FIG. 7 is an identification chromatogram of AP4, FIG. 8 is a mass spectrum of AP4, and the identification molecular weight is 1282g/mol, which is consistent with the theoretical value; FIG. 9 shows an identification chromatogram of AP5, FIG. 10 shows a mass spectrum of AP5, and the identification molecular weight is 1066.6g/mol, which corresponds to the theoretical value. As can be seen from FIGS. 1 to 10, the 5 polypeptides were successfully synthesized.
Example 2 porcine lymphocyte proliferation assay
1. ASFV inactivated virus immunized pigs.
5 male long white pigs of 90 days old were immunized with ASFV epidemic strain inactivated virus (from african swine fever area laboratory of the institute of veterinarian, department of academy of agricultural sciences, china) (10 HID 50), boosted once a month later, euthanized 7 days later, dissected and spleened out. Non-immunized healthy pigs were used as negative control group.
2. And (5) preparing, culturing and detecting proliferation of spleen cells.
1) The collected pig spleens were subjected to aseptic treatment with 75% alcohol, washed three times with PBS, the spleens were cut into small pieces, placed in folded sterile gauze (2 layers), and the spleens were ground in a dish containing 5mL of medium containing serum 1640.
2) The liquid was then aspirated into a 15ml centrifuge tube, 1000rpm, and centrifuged for 5min.
3) The supernatant was discarded and the pellet (cells) was knocked to suspend it uniformly.
4) 10ml of erythrocyte lysate is added for 10min of lysis, then 6ml of medium containing serum 1640 is added for stopping the lysis, and after being evenly mixed, 1000rpm is carried out for 5min of centrifugation.
5) Removing supernatant, knocking precipitate again to suspend it uniformly, diluting 10 μl of suspension 40 times, counting cells, and adjusting cell concentration to 1×10 6 Individual cells/mL.
6) After CFSE staining, the prepared spleen cells were seeded into 24-well plates (1X 10) 6 Individual cells/well).
7) Adding 0.2 mug of the first polypeptide to the fifth polypeptide provided by the invention respectively, and placing the mixture in CO 2 Culturing in an incubator for 60 hours, and detecting the proliferation of ASFV antigen-specific spleen cells by flow cytometry.
The detection results are shown in FIG. 11. FIG. 11 is a flow chart of data statistics for the proliferation of AP 1-AP 5-activated lymphocytes and monocytes. As can be seen from fig. 11, the 5 polypeptides of the present invention can significantly promote proliferation of spleen immune cells specific to ASFV, and the main type of immune cells is monocytes.
Healthy porcine spleen lymphocytes were treated in the same manner and the effect of the polypeptide on non-antigen specific spleen cell proliferation in the porcine organism was examined.
The detection results are shown in FIG. 12. FIG. 12 is a flow chart of data statistics of AP 1-AP 5 promoting proliferation of healthy porcine mononuclear macrophages. From fig. 12, it can be seen that 5 polypeptides of the present invention can also significantly promote proliferation of spleen immune cells in healthy pigs. The 5 polypeptides have the function of broad-spectrum promotion of pig immunity.
Example 3 detection of IFN-y secretion by spleen lymphocytes of different subtypes.
Pig immunization procedure and spleen cell isolation culture were as in example 2. After obtaining dispersed spleen cells, the spleen cells are prepared into single cell suspension with RPMI1640 complete medium, and the concentration is 1 multiplied by 10 6 /ml. Inoculated into 24-well plates, 0.2. Mu.g of the polypeptide provided by the invention was added to each well, and cultured in a CO2 incubator for 60 hours. Collecting cells of each well, labeling cells with specific antibodies of pig CD3, CD4, CD8 and IFN-gamma, washing with PBS buffer solution containing 2% serum for 2 times, dispersing into cell suspension, detecting with flow cytometry, and determining CD4 + T lymphocytes and CD8 + T lymphocytes secrete IFN-gamma levels.
The detection results are shown in FIGS. 13 to 17. Wherein FIG. 13 is a graph of the level of IFN-gamma secretion by AP1 promoting immune cells of different subtypes; FIG. 14 is a graph showing that AP2 promotes IFN-gamma secretion by various subtypes of immune cells; FIG. 15 shows the levels of AP3 promoting secretion of IFN-gamma by various subtypes of immune cells; FIG. 16 shows that AP4 promotes IFN-gamma secretion by various subtypes of immune cells; FIG. 17 is a graph showing that AP5 promotes IFN-gamma secretion by various subtypes of immune cells; from FIGS. 13 to 17, it can be seen that five polypeptides of the present invention significantly enhance IFN-gamma secretion from monocytes.
EXAMPLE 5 polypeptide immunized animal immunocyte typing experiment
3 male white pigs of 90 days old were immunized with a mixture of 5 polypeptides of the invention (100. Mu.g/head of each polypeptide), boosted once a month, and the proportion of immune cells in peripheral blood was measured by a 14d flow cytometer after two-immunization. The same volume of PBS immunized group served as control.
The detection results are shown in FIGS. 18 to 19. Wherein FIG. 18 is a graph showing the proportion level of the sub-population of B lymphocytes in the secondary claim 14 d; FIG. 19 is CD8 + T lymphocyte subpopulation proportion level. From FIGS. 18 to 19, it is apparent that the mixed immunization with 5 polypeptides of the present invention can promote the generation of lymphocyte immune response in swine organism and enhance swine immunity.
EXAMPLE 6 immunopotentiator immune cell typing experiment of animals
3 male white pigs of 90 days old are immunized BY 5 polypeptide mixtures (100 mug/head parts of each polypeptide) of the foot-and-mouth disease O/MYA98/BY/2010 strain inactivated vaccine or inactivated vaccine, one month later, the immunization is enhanced, and the proportion of immune cells in peripheral blood is detected BY a 14d flow cytometer after double immunization. The same volume of PBS immunized group served as control.
The detection results are shown in fig. 20-21, wherein fig. 20 shows the proportion level of the secondary-exemption 14d and the B lymphocyte subpopulation; FIG. 21 is CD8 + T lymphocyte subpopulation proportion level. From FIGS. 20 to 21, it is understood that 5 polypeptides of the present invention can enhance the level of lymphocyte immune response of a swine foot-and-mouth disease vaccine, and enhance the immunogenicity of the vaccine.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Sequence listing
<110> the animal doctor institute of Lanzhou, china academy of agricultural sciences
<120> polypeptide for promoting pig organism to generate broad spectrum immune response and application thereof
<160> 5
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 1
Lys Met Arg Ile Glu Ile Phe Trp Glu Leu
1 5 10
<210> 2
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 2
Arg Ser Met Met Met Val Phe Asn Gln Leu
1 5 10
<210> 3
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 3
Tyr Gly Ile Ala Leu Gln Tyr Val Leu
1 5
<210> 4
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 4
Ile Phe Asn Phe Ile Glu Asn Ile Trp Ser
1 5 10
<210> 5
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 5
Tyr Thr Glu Met Leu Ala Leu Ile Ile
1 5
Claims (5)
1. A polypeptide for promoting a pig organism to generate a broad-spectrum immune response, wherein the polypeptide is a third polypeptide or a fifth polypeptide;
the amino acid sequence of the third polypeptide is shown as SEQ ID NO. 3;
the amino acid sequence of the fifth polypeptide is shown as SEQ ID NO. 5;
the polypeptide promotes the pig body to generate broad-spectrum immune response by promoting the proliferation of lymphocytes and mononuclear macrophages in the pig body.
2. The polypeptide of claim 1, wherein the lymphocytes comprise spleen lymphocytes, peripheral blood T cells, and B cells in peripheral blood.
3. The polypeptide of claim 1, wherein the mononuclear macrophages comprise spleen mononuclear macrophages and peripheral blood mononuclear macrophages.
4. Use of the polypeptide according to any one of claims 1 to 3 for the preparation of a pig immunopotentiator.
5. An immunopotentiator for pigs, the active ingredient of which comprises the polypeptide of any one of claims 1 to 3.
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| CN202111306043.9A CN113929746B (en) | 2020-08-26 | 2020-08-26 | Polypeptide for promoting pig organism to generate broad-spectrum immune response and application thereof |
| CN202010871941.8A CN111925417B (en) | 2020-08-26 | 2020-08-26 | Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof |
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| US20080131449A1 (en) * | 2006-06-22 | 2008-06-05 | Matthias Rath | Polypeptides from African Swine Fever virus as vaccines for preventive and therapeutic use |
| GB201410971D0 (en) * | 2014-06-19 | 2014-08-06 | Pirbright Inst The | Vaccine |
| JP2019507784A (en) * | 2016-03-07 | 2019-03-22 | ヴァージニア テック インテレクチュアル プロパティーズ，インコーポレーテッド | Chimeric porcine circovirus type 2 (PCV2) vaccine |
| CN111393531B (en) * | 2019-01-03 | 2023-01-17 | 浙江海隆生物科技有限公司 | Subunit fusion protein CD2V-Fc and preparation method and application thereof |
| CN111471089B (en) * | 2019-01-24 | 2023-11-24 | 浙江海隆生物科技有限公司 | Recombinant African swine fever virus CD2V subunit protein and preparation method and application thereof |
| CN114044805B (en) * | 2020-08-26 | 2023-10-27 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig organism to generate broad-spectrum immune response and application thereof |
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| CN111925417B (en) | 2021-11-26 |
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| CN114044805A (en) | 2022-02-15 |
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