CN113896772B - Polypeptide for promoting swine organism to generate African swine fever virus antigen specific immune response and application thereof - Google Patents
Polypeptide for promoting swine organism to generate African swine fever virus antigen specific immune response and application thereof Download PDFInfo
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- CN113896772B CN113896772B CN202111307660.0A CN202111307660A CN113896772B CN 113896772 B CN113896772 B CN 113896772B CN 202111307660 A CN202111307660 A CN 202111307660A CN 113896772 B CN113896772 B CN 113896772B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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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
- A61K39/12—Viral antigens
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- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
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- 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
- C12N2710/12022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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- 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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The application provides a polypeptide for promoting a swine organism to generate african swine fever virus antigen specific immune response and application thereof, belonging to the technical field of biological medicine; 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 obviously promote proliferation of ASFV sensitized immune cells, and can promote the pig organism to generate an ASFV antigen-specific immune response by promoting secretion of IFN-gamma by pig mononuclear macrophages and B, T lymphocytes. After the animals are immunized, the immune response of the pig organism can be obviously promoted. The polypeptide or the polypeptide polymer obtained by polymerizing the polypeptide can be used for preparing subunit vaccines of African swine fever virus.
Description
The application relates to a polypeptide for promoting swine fever virus antigen specific immune response of pig organisms and application thereof, which are classified as a polypeptide for promoting the swine fever virus antigen specific immune response of pigs on the basis of 26 th month of 2020 and the application number of CN 202010873520.9.
Technical Field
The application relates to the technical field of biomedicine, in particular to a polypeptide for promoting a swine organism to generate an African swine fever virus antigen specific immune response and application thereof.
Background
African swine fever (African Swine fever, ASF) is an acute, hemorrhagic, virulent infectious disease caused by infection of domestic pigs and various wild pigs (e.g., african wild pigs, european wild pigs, etc.) with African swine fever virus (African Swinefever virus, ASFV). African swine fever virus mainly infects immune cells such as macrophages and monocytes, and ASFV escapes from the host's innate and inherent immune defense systems through different mechanisms, such as type I Interferon (IFN) response, apoptosis, inflammatory response and activation of specific target genes.
Vaccine is one of the most effective methods for preventing african swine fever. Traditional live attenuated vaccines and inactivated vaccines are currently the most commonly used vaccines. However, natural attenuated strains or attenuated strains due to gene deletion of ASFV have various defects of biological potential safety hazard, immune side reaction, toxicity scattering danger, unclear genetic background and the like, so that the application of the natural attenuated strains or attenuated strains in vaccine production is limited. ASFV encodes up to 167 proteins, which makes it very difficult to screen candidate antigens that can induce immune protection in the body. However, several proteins have been reported as targets for virus neutralization and recombinant protein vaccine studies have been performed. Both P30 and P54 proteins are located outside the virus and are involved in virus adsorption and internalization, respectively. Immunization of pigs with p30 and p54 proteins induces the production of neutralizing antibodies, but is not resistant to lethal challenge, and the course of the disease is unchanged, whereas immunization with a combination p30 and p54 vaccine produces neutralizing antibodies, which also improve the course of the disease. Inova et al evaluated 46 peptides mimicking viral proteins to determine their ability to establish protective immune responses. Vaccination with certain combination peptides may delay home pig death. However, no fully effective ASFV subunit vaccine has been reported so far.
Disclosure of Invention
The application aims to provide a polypeptide for promoting a swine organism to generate an African swine fever virus antigen specific immune response and application thereof, the polypeptide can be used for preparing an ASFV subunit vaccine, and the prepared subunit vaccine can effectively enhance the organism ASFV specific immune response and prevent African swine fever virus infection.
In order to achieve the above object, the present application provides the following technical solutions:
the application provides a polypeptide for promoting a swine organism to generate an African swine fever virus antigen specific 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 swine organism to generate an African swine fever virus antigen-specific immune response by promoting the secretion of IFN-gamma by swine mononuclear macrophages and B, T lymphocytes; and/or the number of the groups of groups,
the polypeptide promotes the pig organism to generate the African swine fever virus antigen specific immune response by promoting the proliferation of ASFV antigen specific mononuclear macrophages.
Preferably, the ASFV antigen-specific monocyte macrophage comprises CD4 + T lymphocytes, CD8 + Proliferation of T lymphocytes and B cells.
Preferably, the african swine fever virus antigen-specific lymphocytes comprise CD4 + T lymphocytes, CD8 + T lymphocytes and B cells.
The application provides a polypeptide polymer obtained by polymerizing the polypeptide in the scheme.
The application 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 application provides a subunit vaccine of African swine fever virus, and an active ingredient of the subunit vaccine comprises the polypeptide or the polypeptide polymer in the scheme.
Preferably, the dosage form of the subunit vaccine comprises an injection.
Preferably, the content of the polypeptide or the polypeptide polymer in the subunit vaccine is 100 mug/component/head.
The application has the beneficial effects that: the application provides a polypeptide for promoting a swine organism to generate an African swine fever virus antigen specific 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 of the application can obviously promote the proliferation of ASFV sensitized immune cells, and can promote the pig organism to generate an ASFV antigen specific immune response by promoting the secretion of IFN-gamma by pig mononuclear macrophages and B, T lymphocytes. After the animals are immunized, the immune response of the pig organism can be obviously promoted. The polypeptide or the polypeptide polymer obtained by polymerizing the polypeptide can be used for preparing subunit vaccines of African swine fever virus. The polypeptide provided by the application has no toxic or side effect and biological potential safety hazard, and has an industrialization advantage as an ASFV vaccine component.
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.
Detailed Description
The application provides a polypeptide for promoting a swine organism to generate an African swine fever virus antigen specific 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 swine organism to generate an African swine fever virus antigen-specific immune response by promoting the secretion of IFN-gamma by swine mononuclear macrophages and B, T lymphocytes; and/or the polypeptide promotes the pig body to generate the African swine fever virus antigen-specific immune response by promoting proliferation of ASFV antigen-specific mononuclear macrophages.
In the present application, the African swine fever virus antigen-specific lymphocytes comprise CD4 + T lymphocytes, CD8 + T lymphocytes and B cells.
In the application, the amino acid sequence of the first polypeptide is shown as SEQ ID NO.1, and specifically comprises the following steps: SMAAKIFIV (Ser-Met-Ala-Ala-Lys-Ile-Phe-Ile-Val) has an average molecular weight of 979.23g/mol and a chemical formula: c (C) 46 H 78 N 10 O 11 S, S. The theoretical isoelectric point of the polypeptide is pH 10.09, the GRAVY value of the polypeptide is 1.87, and the polypeptide has hydrophobicity. This property favors its interaction with host proteinsOr cell surface binding, can more effectively initiate an immune response.
In the application, the amino acid sequence of the second polypeptide is shown as SEQ ID NO.2, and specifically comprises the following steps: STQAYNDFL (Ser-Thr-Gln-Ala-Tyr-Asn-Asp-Phe-Leu). The average molecular weight is 1058.09g/mol, and the chemical formula is: c (C) 47 H 67 N 11 O 17 . The theoretical isoelectric point of the polypeptide is pH 3.12, the GRAVY value of the polypeptide is-0.54, and the polypeptide has hydrophilicity. In the prior art, no similar compound is found.
In the application, the amino acid sequence of the third polypeptide is shown as SEQ ID NO.3, and specifically comprises the following steps: FQMNVSACAW (Phe-Gln-Met-Asn-Val-Ser-Ala-Cys-Ala-Trp). The average molecular weight is 1156.33g/mol, and the chemical formula is: c (C) 51 H 73 N 13 O 14 S 2 . The polypeptide has a theoretical isoelectric point of pH 5.25, a GRAVY value of 0.63, hydrophobicity, and can be combined with host protein or cell surface to effectively start immune response.
In the application, the amino acid sequence of the fourth polypeptide is shown as SEQ ID NO.4, and specifically comprises the following steps: PPTQRVDPA (Pro-Pro-Thr-Gln-Arg-Val-Asp-Pro-Ala). The average molecular weight is 980.07g/mol, and the chemical formula is: c (C) 42 H 69 N 13 O 14 . The theoretical isoelectric point of the polypeptide is pH 7, the GRAVY value of the polypeptide is-1.22, and the polypeptide has hydrophilicity.
In the application, the amino acid sequence of the fifth polypeptide is shown as SEQ ID NO.5, and specifically comprises the following steps: NSTRFTTEDP (Asn-Ser-Thr-Arg-Phe-Thr-Thr-Glu-Asp-Pro). The average molecular weight is 1167.17g/mol, and the chemical formula is: c (C) 48 H 74 N 14 O 20 . The theoretical isoelectric point of the polypeptide is pH 4.07, the GRAVY value of the polypeptide is-1.67, and the polypeptide has hydrophilicity.
In the application, 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.
The application provides a polypeptide polymer obtained by polymerizing the polypeptide in the scheme.
The application 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 application provides a subunit vaccine of African swine fever virus, and an active ingredient of the subunit vaccine comprises the polypeptide or the polypeptide polymer in the scheme.
Preferably, the dosage form of the subunit vaccine comprises an injection.
Preferably, the subunit vaccine has a content of the polypeptide or polypeptide polymer of 100 μg/component/head.
In the present application, the polypeptide is preferably synthesized by Shanghai Biotechnology Co., ltd.
The present application provides a polypeptide polymer comprising the polypeptide of the above scheme.
The application 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.
In the present application, the polypeptide or the polypeptide polymer is prepared by promoting CD4 in pig body + CD8 + The T cells secrete IFN-gamma to promote the swine organism to generate african swine fever virus antigen specific immune response; alternatively, the polypeptide or the polypeptide polymer promotes swine organism to generate african swine fever virus antigen-specific immune response by promoting proliferation of ASFV antigen-specific mononuclear macrophages; the ASFV antigen-specific monocyte macrophage preferably includes CD4 + T lymphocytes, CD8 + Proliferation of T lymphocytes and B cells.
The application provides a subunit vaccine of African swine fever virus, and an active ingredient of the subunit vaccine comprises the polypeptide of the scheme or the polypeptide polymer of the scheme. The subunit vaccine provided by the application can enhance the immunity of a pig organism.
The preparation according to the scheme or the subunit vaccine according to the scheme respectively comprise injection; the content of the polypeptide or the polypeptide polymer in the preparation or the subunit vaccine is 100 mug/component/head, namely the content of each polypeptide or each polypeptide polymer is 100 mug/head; the formulation or the subunit vaccine preferably further comprises an adjuvant 50V2; the state of the formulation or the subunit vaccine is preferably a water-in-oil emulsified state.
The technical solutions of the present application will be clearly and completely described in the following in connection with the embodiments of the present application. It will be apparent that the described embodiments are only some, but not all, embodiments of the application. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
Example 1 solid phase Synthesis and purity detection of Polypeptides
The polypeptides of the application are as follows:
SEQ ID NO.1：SMAAKIFIV，AP1；
SEQ ID NO.2：STQAYNDFL，AP2；
SEQ ID NO.3：FQMNVSACAW，AP3；
SEQ ID NO.4：PPTQRVDPA，AP4；
SEQ ID NO.5：NSTRFTTEDP，AP5；
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 979.55g/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 1058.5g/mol, which is consistent with the theoretical value; FIG. 5 is an identification chromatogram of AP 3; FIG. 6 is a mass spectrum of AP3, with a molecular weight of 1156.45g/mol, consistent with theory; FIG. 7 is an identification chromatogram of AP 4; FIG. 8 is a mass spectrum of AP4, with a molecular weight of 979.9g/mol, consistent with theory; FIG. 9 is an identification chromatogram of AP 5; FIG. 10 is a mass spectrum of AP5, with an identified molecular weight of 1167.95g/mol, consistent with theory; 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, institute of veterinarian, academy of agricultural sciences, china) (10 HID 50), boosted once a month, euthanized 7 days later, dissected and spleened. 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 polypeptide (AP 1-AP 5) provided by the application into 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, each of the 5 polypeptides of the present application significantly promoted proliferation of ASFV-specific spleen immune cells, the main type of immune cells being lymphocytes and 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 the proliferation of healthy porcine lymphocytes and mononuclear macrophages promoted by AP1 to AP 5. As can be seen from fig. 12, the 5 polypeptides of the present application do not significantly promote proliferation of spleen immune cells in healthy pigs.
Example 3 detection of IFN-gamma 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. Inoculating into 24 hole plate, adding 0.2 mug of polypeptide (AP 1-AP 5) provided by the application into each hole, placing into CO 2 Culturing in an 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 B lymphocyte and CD4 respectively + T lymphocytes, CD8 + T lymphocytes, CD4 + CD8 + T lymphocytes and monocytes macrophages 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 application significantly enhance IFN-gamma secretion from monocytes and B, T lymphocytes.
EXAMPLE 4 polypeptide immunized animal immunocyte typing experiment
3 male white pigs of 90 days old were immunized with a mixture of 5 polypeptides of the application (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 application can promote the generation of lymphocyte immune response in swine organism and enhance swine immunity.
The foregoing is merely a preferred embodiment of the present application 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 application, which are intended to be comprehended within the scope of the present application.
Sequence listing
<110> the animal doctor institute of Lanzhou, china academy of agricultural sciences
<120> polypeptide for promoting swine organism to generate African swine fever virus antigen specific immune response and application thereof
<160> 5
<170> SIPOSequenceListing 1.0
<210> 1
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 1
Ser Met Ala Ala Lys Ile Phe Ile Val
1 5
<210> 2
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 2
Ser Thr Gln Ala Tyr Asn Asp Phe Leu
1 5
<210> 3
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 3
Phe Gln Met Asn Val Ser Ala Cys Ala Trp
1 5 10
<210> 4
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 4
Pro Pro Thr Gln Arg Val Asp Pro Ala
1 5
<210> 5
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 5
Asn Ser Thr Arg Phe Thr Thr Glu Asp Pro
1 5 10
Claims (4)
1. A polypeptide for promoting swine organism to generate african swine fever virus antigen specific 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.
2. Use of the polypeptide of claim 1 for the preparation of a formulation for promoting an african swine fever virus antigen-specific immune response in a swine organism.
3. A subunit vaccine of african swine fever virus, the active ingredient of which comprises the polypeptide of claim 1.
4. A subunit vaccine according to claim 3 wherein the dosage form of the subunit vaccine comprises an injectable formulation.
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| CN202111307660.0A CN113896772B (en) | 2020-08-26 | 2020-08-26 | Polypeptide for promoting swine organism to generate African swine fever virus antigen specific immune response and application thereof |
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| CN202111307660.0A CN113896772B (en) | 2020-08-26 | 2020-08-26 | Polypeptide for promoting swine organism to generate African swine fever virus antigen specific immune response and application thereof |
| CN202010873520.9A CN112209996B (en) | 2020-08-26 | 2020-08-26 | Polypeptide for promoting swine organisms to generate African swine fever virus antigen specific immune response and application thereof |
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| CN202111307559.5A Active CN113896771B (en) | 2020-08-26 | 2020-08-26 | Polypeptide for promoting swine organism to generate African swine fever virus antigen specific immune response and application thereof |
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| CN110551695A (en) * | 2019-08-14 | 2019-12-10 | 中国农业科学院上海兽医研究所(中国动物卫生与流行病学中心上海分中心) | African swine fever virus four-gene deletion low virulent strain 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 |
| CN109195623A (en) * | 2016-03-07 | 2019-01-11 | 弗吉尼亚科技知识产权有限公司 | 2 type of chimeric porcine circovirus type (PCV2) vaccine |
| CN110904127A (en) * | 2018-09-18 | 2020-03-24 | 瓦赫宁恩研究基金会 | African swine fever virus vaccine |
| CN111018996A (en) * | 2019-10-31 | 2020-04-17 | 河南省生物工程技术研究中心 | Neutralizing epitope subunit vaccine for African swine fever |
| CN111018995A (en) * | 2019-10-31 | 2020-04-17 | 河南省生物工程技术研究中心 | B, T cell epitope tandem fusion vaccine for African swine fever |
| CN110862435B (en) * | 2019-12-05 | 2021-09-24 | 中国农业大学 | African swine fever CTL epitope polypeptide and application thereof |
| CN111004330A (en) * | 2019-12-13 | 2020-04-14 | 天津大学 | Method for preparing African swine fever virus P30 and P54 yeast vaccines |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110551695A (en) * | 2019-08-14 | 2019-12-10 | 中国农业科学院上海兽医研究所(中国动物卫生与流行病学中心上海分中心) | African swine fever virus four-gene deletion low virulent strain and application thereof |
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| CN113896772A (en) | 2022-01-07 |
| CN112209996B (en) | 2021-11-26 |
| CN113896771A (en) | 2022-01-07 |
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| CN113896771B (en) | 2023-10-27 |
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