CN108117583A - A kind of immunopotentiator and its application - Google Patents
A kind of immunopotentiator and its application Download PDFInfo
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Abstract
The present invention provides a kind of immunopotentiator and the foot and mouth disease vaccine composition containing the immunopotentiator is with applying.The immunopotentiator includes three swine foot-and-mouth disease virus CTL epitope polypeptides, any of the above combination of one or both of polypeptide in the polypeptide and sequence table in the polypeptide, sequence table as in sequence table shown in sequence 1 shown in sequence 2 shown in sequence 3.The foot and mouth disease vaccine composition includes immunopotentiator and Schweineseuche antigen.Immunopotentiator of the invention can largely be prepared by solid-state chemical reaction method method; it is mixed with inactivation antigen with certain proportion; vaccine combination is prepared convenient for the production of scale; the vaccine combination both can induce body and generate humoral immune response level; also it is horizontal that the cellullar immunologic response of body can be enhanced, and preferable immune effect can be obtained.The immunopotentiator of the present invention is safe and reliable, has broad application prospects.
Description
Technical field
The invention belongs to veterinary biologics technical fields, specifically, the present invention relates to a kind of immunopotentiator and contain
The foot and mouth disease vaccine composition of the immunopotentiator and application.
Background technology
Aftosa (Foot and Mouth Disease, FMD) is by foot and mouth disease virus (Foot-and-Mouth
Disease virus, FMDV) caused by a kind of acute, hot, high degree in contact infectiousness for occurring of the artiodactyls such as pig, ox, sheep
With the deadly infectious disease of quick long-distance communications, with infectiousness is strong, transmission speed is fast, harm is serious and famous, the disease
Outbreak of epidemic gives the animal husbandry for delivering area to cause huge economic loss (Chang Huiyun, 2017).World Organization for Animal Health
(OIE) many animals that being classified as must circulate a notice of suffer from one of infectious disease altogether, and China is classified as a kind of zoonosis.
Aftosa infectiousness is high, propagates rapidly, the death of cub, adults are often resulted in after the livestocks such as infected pigs, ox, sheep
Growth ability drastically declines after infection.The disease spread speed is rapid, can cause it is global be very popular, not only animal husbandry is made
Into huge economic loss, and serious international trade and the reputation for influencing animal and its livestock products.
FMDV belongs to Picornaviridae, is single strand plus RNA virus, because its genome has the variability of height, causes
FMDV, which constantly evolves, generates new virus subtype, and difficulty is brought to the prevention and control and purification of FMD.FMD is to be related to national economy
The great animal epidemic of safety.At present, most developing countries and area are all controlled by being inoculated with the method for inactivated vaccine
The prevalence of FMD, and traditional inactivated vaccine mainly induces body to generate humoral immune response, lacks cellullar immunologic response.In addition,
With the continuous innovation of antigen purification technology, FMD subunit vaccines, virus-like particle (VLP) vaccine and synthetic peptide vaccine etc.
New generation vaccine will largely occur, although the security of vaccine is improved, but its immunogenicity is often not satisfactory, and lack equal
The humoral immune response and cellullar immunologic response of weighing apparatus, it is therefore desirable to develop safer efficient immunopotentiator to improve vaccine
Cellullar immunologic response it is horizontal.
Epitope refers to that the knot of specificity can occur with T cell receptor, B-cell receptor or antigen-binding fragment for antigen
It closes, there is special construction and immunocompetent chemical group, be the material base for causing immune response.B cell compared to FMD
Epitope, Th cell epitopes, it is also less for the research of cytotoxic T cell CTL epitopes at present, there is research to confirm FMDV specificity
CTL epitopes can induce body generate cellular immunity response, therefore develop CTL epitopes immunopotentiator for cell-mediated
Anti- FMDV be immunized with important application value.
The content of the invention
The object of the present invention is to provide a kind of immunopotentiator and the foot and mouth disease vaccine containing the immunopotentiator combines
Object and application.The foot and mouth disease vaccine composition not only can induce body and generate humoral immune response, but also can enhance body and generate carefully
Born of the same parents' immune response can obtain preferable immune effect.
Immunopotentiator provided by the invention, as shown in the polypeptide shown in sequence in sequence table 1, sequence 2 in sequence table
The peptide composition composition of one or both of polypeptide in polypeptide and sequence table shown in sequence 3 any of the above combination.
When the immunopotentiator is as the polypeptide and sequence shown in the polypeptide shown in sequence in sequence table 1, sequence 2 in sequence table
During two kinds in the polypeptide in list shown in sequence 3 compositions, their weight fraction ratio is 0.5-1： 0.5-1；
When the immunopotentiator is as the polypeptide and sequence shown in the polypeptide shown in sequence in sequence table 1, sequence 2 in sequence table
When polypeptide in list shown in sequence 3 forms；It is more shown in sequence 2 in polypeptide, sequence table in the sequence table shown in sequence 1
The ratio of weight and number of polypeptide in peptide or sequence table shown in sequence 3 is 0.5-1： 0.5-1：0.5-1.Preferably, the sequence table
The parts by weight of polypeptide in polypeptide or sequence table in polypeptide, sequence table shown in middle sequence 1 shown in sequence 2 shown in sequence 3
Than for 1：1：1.
Wherein, 3 institute of sequence in the polypeptide in sequence table shown in sequence 1, the polypeptide in sequence table shown in sequence 2 or sequence table
The polypeptide shown is the polypeptide for Immune-enhancing effect, falls within protection scope of the present invention.
Foot and mouth disease vaccine composition provided by the invention, including above-mentioned immunopotentiator.
Specifically, the vaccine combination includes：
(1) above-mentioned immunopotentiator,
With (2) Schweineseuche antigen.
Wherein, the vaccine combination is immunopotentiator and the mixture of foot and mouth disease vaccine.
The Schweineseuche antigen is Schweineseuche inactivation antigen, swine foot-and-mouth disease virus sample particulate antigen, Schweineseuche are sub-
Unit antigen or Schweineseuche polypeptide antigen；Wherein, Schweineseuche inactivation antigen is selected from Schweineseuche O-shaped and A type vaccine virus
Strain.Preferably, the pig FMDV inactivation antigens are selected from O/Mya98/XJ/2010 plants of pig, O/GX/09-7 plants, AF/72 plants, A/
The inactivation of viruses antigen of one or more kinds of any combination in QH/09 plants, is most preferably O/Mya98/XJ/2010 plants.
When preparing foot and mouth disease vaccine composition, by the immunopotentiator and pig FMDV inactivation antigens in buffer solution system
In carry out incubation mixing, buffer solution system is preferably 20mMTris-HCl (pH 8.0) solution containing 50mMNaCl, the incubation
Condition is preferably 4 DEG C of overnight incubations.
Preferably, in the foot and mouth disease vaccine composition, the ratio of weight and number of immunopotentiator and Schweineseuche antigen
It can determine that weight fraction ratio is specifically as follows 0.1-10 according to antigen type, immunizing dose etc.：1st, preferably 1-5：1.
In an embodiment of the invention, when antigen selects Schweineseuche inactivation antigen, immunopotentiator and pig
The weight fraction ratio of aftosa is 3：1.
Further, the foot and mouth disease vaccine composition further includes veterinarily acceptable adjuvant, and of the invention is another
One is designed to provide a kind of foot and mouth disease vaccine composition, including the immunopotentiator, Schweineseuche antigen and animal doctor
Acceptable adjuvant on.
The immunizing dose of the Schweineseuche composition of the present invention can according to the main species by immunized animal of immune swine,
Strain, age, weight, health status etc. determine, for example, can be 1-300ug Schweineseuches composition/head part；Such as when above-mentioned
Schweineseuche antigen be Schweineseuche inactivation antigen when, can according to the species of inactivation antigen set immunizing dose 1-200ug pigs
Foot-and-mouth disease antigen/head part pig.
In one embodiment of the invention, Schweineseuche inactivation antigen be O/Mya98/XJ/2010 plants of inactivation antigens, pig
The immunizing dose of pig FMDV inactivation antigens is 10ug/ parts in aftosa composition, and the immunizing dose of immunopotentiator is 30 μ g/
Head part.
Term used herein " foot and mouth disease vaccine composition " refers to can be used for prevention by pig FMDV infection institute cardinal symptom
Or the vaccine of disease, the vaccine combination may include any vaccine that can be used in prevention pig and infected from pig FMDV.
Term of the present invention " immune amount " refers to the immunizing dose for pig FMD vaccine combinations, immunizing dose it is big
It is small mainly whether phase to be immunized before by the species of immunized animal, strain, age, weight, health status and the animal
The factors such as the vaccine with strain influence.
Term " adjuvant " of the present invention may include water-in-oil emulsion, oil in water emulsion, W/O/W emulsion, aluminium glue assistant
One or more of agent, saponin(e or Gel adjuvants；Preferably, the adjuvant is 206 adjuvants of Montanide ISA.The assistant
Agent is 1 with the volume ratio of the immunopotentiator and Schweineseuche inactivation antigen mixture:1.
The immunopotentiator of the present invention can largely be prepared by solid-state chemical reaction method method, with inactivation antigen with certain
Ratio mixes, and convenient for the production of scale, both can induce body and has generated humoral immune response level, and can also enhance the cell of body
Immune response is horizontal, and can obtain preferable immune effect.It is safe and reliable, there is more wide application prospect.
Description of the drawings
The ELISPOT result of the tests of rear 14th day IFN-γ are immunized in Fig. 1
The ELISPOT result of the tests of rear 21st day IFN-γ are immunized in Fig. 2
The ELISPOT result of the tests of rear 14th day IFN-γ are immunized in Fig. 3
The ELISPOT result of the tests of rear 21st day IFN-γ are immunized in Fig. 4
Specific embodiment
Illustrate the present invention referring to specific embodiment.It will be appreciated by those skilled in the art that these embodiments are only
For illustrating the present invention, do not limit the scope of the invention in any way.
Experimental method in following embodiments is conventional method unless otherwise specified.Medicine used in following embodiments
Material raw material, reagent material etc. unless otherwise specified, are commercially available products.
Term used herein " CTL epitope polypeptides " refers to cytotoxic T cell epitope polypeptide (Cytotoxic T
Lymphocytesepitope, abbreviation CTL epitope polypeptide), target antigen is handled through antigen presenting cell with " Antigenic Peptide-MHC-I classes
The form of molecule " compound present to antigen presenting cell or target cell surface after can by CD8+T cell recognitions, accordingly with
The Antigenic Peptide that MHC-I quasi-molecules combine is CTL epitope polypeptides.CTL epitope polypeptides being capable of activation-inducing generation specific cell
Toxic T lymphocyte (CTL cells), induction body generate cellular immunity response.
Term used herein " CTL epitope polypeptides " refers to that being located at FMDV structural proteins has the function of identical sequence or have
The peptide fragment of suitable segment.
The screening of embodiment 1, CTL epitope polypeptides
After predicting polypeptide immune body, by the polypeptide antigen of MHCI quasi-molecule submissions, body can be stimulated to generate Memorability T thin
Born of the same parents.When polypeptide antigen stimulates T cell again, memory t cell can induce generation cell factor.By to cytokine levels
Detection and using biometric analysis can to prediction polypeptide whether be CTL epitope polypeptides carry out evaluation analysis.
1. predict polypeptide：
By 10 prediction polypeptide ZM1501~ZM15010 of website prediction and chemical synthesis, sequence information is shown in sequence table.It utilizes
20mMTris-HCl (pH 8.0), 50mMNaCl buffer solutions dissolve prediction polypeptide respectively, and 10 prediction polypeptides are configured to dense
It spends for 1mg/ml solution.Each prediction polypeptide presses 1 according to immunizing dose for g/ parts of 10 μ and 61 adjuvants:1 volume ratio carries out mixing and exempts from
Epidemic disease animal.
Sequence 1 is the amino acid sequence of polypeptide ZM1501 in sequence table；
Sequence 2 is the amino acid sequence of polypeptide ZM1502 in sequence table；
Sequence 3 is the amino acid sequence of polypeptide ZM1503 in sequence table；
Sequence 4 is the amino acid sequence of polypeptide ZM1504 in sequence table；
Sequence 5 is the amino acid sequence of polypeptide ZM1505 in sequence table；
Sequence 6 is the amino acid sequence of polypeptide ZM1506 in sequence table；
Sequence 7 is the amino acid sequence of polypeptide ZM1507 in sequence table；
Sequence 8 is the amino acid sequence of polypeptide ZM1508 in sequence table；
Sequence 9 is the amino acid sequence of polypeptide ZM1509 in sequence table；
Sequence 10 is the amino acid sequence of polypeptide ZM15010 in sequence table.
2. animal is immune：
Experimental animal：The susceptible piglet of 6-7 week old health, Liquid-phase blocking ELISA antibody titer≤1:8.
The present invention detects Liquid-phase blocking ELISA using aftosa Liquid-phase blocking ELISA kit (Lanzhou veterinary institute)
Antibody titer.
Separation, extraction pig PMBC, if pig PMBC can induce when being stimulated through a certain item prediction polypeptide generates higher cell
The factor, compared with control group and other prediction polypeptides, statistical analysis significant difference, then the prediction polypeptide is CTL epitope polypeptides.Such as
The following table 1：Immune grouping is carried out according to the table.The content of wherein immune peptide is g/ parts of 10 μ.It is 2 times immune, after first immunisation
It carries out within 14th day being immunized for second.Each group sets as shown in table 1.
Table 1, immune grouping and processing method
| Group | Group name | Antigen forms | Antigenic content | Adjuvant | Immunization ways | Number of animals | Immunizing dose |
| First group | ZM1501 | ZM1501 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| Second group | ZM1502 | ZM1502 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| 3rd group | ZM1503 | ZM1503 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| 4th group | ZM1504 | ZM1504 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| 5th group | ZM1505 | ZM1505 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| 6th group | ZM1506 | ZM1506 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| 7th group | ZM1507 | ZM1507 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| 8th group | ZM1508 | ZM1508 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| 9th group | ZM1509 | ZM1509 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| Tenth group | ZM15010 | ZM15010 | 10 parts of μ g/ | 61 adjuvants | Former times 5 | 5 | 2ml |
| Control group | -- | -- | -- | -- | -- | 2 | -- |
(1) separation of pig peripheral blood mononuclearcell (PBMC)
A. the new blood of pig is gathered in the 14th day, the 21st day vena cava anterior blood collection method respectively after being immunized, heparin is put into and resists
In solidifying heparin tube.
B. the new blood of 10ml is taken, 1 is pressed with pH 7.4PBS or 0.9%NaCl:After 1 dilution proportion, it is added to that rewarming is extremely
In the 10ml pig lymphocyte separating liquids of room temperature (20 DEG C or so), action is soft during sample-adding, and it is thin to be added slowly to lymph along tube wall
In born of the same parents' separating liquid.20 DEG C, 2000rpm (700g) centrifugations 20min.
C. top layer is blood plasma and dilution after centrifuging, and tube bottom is red blood cell and granulocyte layer, and middle level is lymphocyte point
Chaotropic (canescence cloud and mist layer, the layer are mainly PBMC) draws buffy coat, until in the test tube of a 10ml.2500rpm is centrifuged
10min abandons supernatant.
D. the RPMI 1640 for adding in 5ml dilutes, 1500rpm, centrifugation 10min；It repeats this operation once, removes blood as far as possible
Platelet and anti-coagulants.Cell count is 2 × 10 with streaming reaction solution adjustment cell concentration6A/mL is for use.
(2) ELISPOT is tested
PBMC is added to according to grouping in pre-coated ELISPOT plates in that step (1), sets positive control, feminine gender right
According to blank control.Stimulant is 10 prediction polypeptides, is operated according to kit specification, and the PBMC that every peptide stimulates is done
2 multiple holes.After polypeptide stimulates PBMC, stimulated with this group of animal PBMC without polypeptide as control, what is generated is stimulated to each polypeptide
IFN-γ is compared, and the protein level of the IFN-γ of animal can be dramatically increased between statistical analysis polypeptide, this polypeptide is determined as sun
Property epitope peptide.
(3) analysis result is as follows：
A.ELISPOT result of the tests：It is illustrated in fig. 1 shown below, figure column a, b difference letter, letter is identical to represent statistics
It is not notable to analyze difference, letter is different to represent statistical analysis significant difference.It is more with control group and other predictions 14th day after immune
Peptide is compared, and after polypeptide ZM1501, ZM1502 or ZM1503 stimulate Swine PBMC, can generate the IFN-γ of higher level, statistical
Analyse significant difference；As shown in Fig. 2,21 days after being immunized, the IFN-γ that polypeptide ZM1501, ZM1502 and ZM1503 are generated is higher than equal
Control group and other polypeptides, statistical analysis significant difference.
In conclusion being screened by the animal immune experiment to 10 prediction polypeptides, ZM1501, ZM1502 are finally obtained
Amount to three function CTL epitope polypeptides with ZM1503.This three CTL epitope polypeptides can be used as immunopotentiator to improve Schweineseuche
The immunity of vaccine.
The application of embodiment 2, the vaccine combination of epitope polypeptide containing CTL
1. obtain 3 function CTL epitope polypeptides by being screened in embodiment 1：ZM1501, ZM1502 and ZM1503 are dissolved in
20mMTris-HCl (pH 8.0), 50mMNaCl buffer solutions.3 CTL epitope polypeptides are configured to concentration as 1mg/ml solution.
Wherein,
ZM1501 sequences：SADPVTATV (sequence 1 in sequence table)
ZM1501 sequences：ALDNTTNPT (sequence 2 in sequence table)
ZM1501 sequences：LM QTPSHTL (sequence 3 in sequence table)
Each CTL epitope polypeptides are taken according to immunizing dose and immune grouping or are configured to mixed solution and the O/ containing immunizing dose
Mya98/XJ/2010 inactivation antigens mix, 4 DEG C of overnight incubations, mixed solution and 206 adjuvants of Montanide ISA of acquisition
It is mixed with to obtain foot and mouth disease vaccine composition.
As shown in table 2, the preparation method of each group foot and mouth disease vaccine composition is：
1) inactivated vaccine group：O/Mya98/XJ/2010 inactivation antigens be dissolved in 20mMTris-HCl (pH 8.0),
50mMNaCl buffer solutions are made 1mg/ml solution, take 10ul, are buffered with 20mMTris-HCl (pH 8.0), 50mMNaCl molten
Liquid is diluted to 1ml, then with 206 adjuvants of Montanide ISA according to volume ratio 1：1 mixing.
2) immunopotentiator group 1：By the 1mg/mlZM1501 polypeptide solutions of above-mentioned preparation, 30ul and 10ugO/ are taken
Mya98/XJ/2010 inactivation antigen mixings are diluted to 1ml with 20mMTris-HCl (pH 8.0), 50mMNaCl buffer solutions,
Then with 206 adjuvants of Montanide ISA according to volume ratio 1：1 mixing.
3) immunopotentiator group 2：By the 1mg/mlZM1502 polypeptide solutions of above-mentioned preparation, 30ul and 10ugO/ are taken
Mya98/XJ/2010 inactivation antigen mixings are diluted to 1ml with 20mMTris-HCl (pH 8.0), 50mMNaCl buffer solutions,
Then with 206 adjuvants of Montanide ISA according to volume ratio 1：1 mixing.
4) immunopotentiator group 3：By the 1mg/mlZM1503 polypeptide solutions of above-mentioned preparation, 30ul and 10ugO/ are taken
Mya98/XJ/2010 inactivation antigen mixings are diluted to 1ml with 20mMTris-HCl (pH 8.0), 50mMNaCl buffer solutions,
Then with 206 adjuvants of Montanide ISA according to volume ratio 1：1 mixing.
5) immunopotentiator group 4：The 1mg/mlZM1501 polypeptide solutions of above-mentioned preparation, 1mg/mlZM1502 polypeptides is molten
Liquid, 1mg/mlZM1503 polypeptide solutions respectively take 10ul and 10ugO/Mya98/XJ/2010 inactivation antigen mixings, use
20mMTris-HCl (pH 8.0), 50mMNaCl buffer solutions are diluted to 1ml, are then pressed with 206 adjuvants of Montanide ISA
According to volume ratio 1：1 mixing.
Experimental animal：The susceptible piglet of 6-7 week old health, Liquid-phase blocking ELISA antibody titer≤1:8.
The present invention detects Liquid-phase blocking ELISA using aftosa Liquid-phase blocking ELISA kit (Lanzhou veterinary institute)
Antibody titer.
Animal is immunized：
Such as the following table 2：Immune grouping is carried out according to the table.Wherein g/ parts of 10 μ of inactivation antigen and immunopotentiator is immune
Content is g parts of 30 μ.It is 2 times immune, it carries out second within the 14th day after first immunisation and is immunized.Inactivation antigen is O/Mya98/XJ/
2010 inactivation antigens.Each group sets as shown in table 2.
Table 2, immune grouping and processing method
(1) ELISA of the separation of serum and antibody level is detected.
Divide respectively in the 14th day, 21 days, 35 days, 49 days, 2 months, March, April, May, June, July, August blood sampling after immune
From serum, the detection of antibody level is carried out.
(2) new blood of pig is gathered in the 14th day, the 21st day vena cava anterior blood collection method respectively after being immunized, according to embodiment
Method carries out the separation of pig peripheral blood mononuclearcell (PBMC), ELISPOT experiments in 1.
(3) analysis result is as follows：
A. antibody level testing result：
It is as shown in table 3 below：Each time point carries out ELISA testing results, immunopotentiator 1-3 to antibody level after immune
Group, pig entirety antibody level and mean antibody levels are slightly above inactivated vaccine group, but are below 4 groups of immunopotentiator；It is immune
4 groups of pig entirety antibody levels of reinforcing agent and mean antibody levels are higher than inactivated vaccine group, and the antibody duration is longer, explanation
By 3 polypeptides in combination using can be remarkably reinforced body be directed to inactivation antigen antibody level.
Table 3, antibody level testing result
B.ELISPOT result of the tests：It is illustrated in fig. 3 shown below, the 14th day after being immunized, figure column a, b, c difference letter,
The identical expression statistical analysis difference of letter is not notable, containing the identical expression statistical analysis significant difference of a letter.Letter is complete
Complete different expression statistical analysis differences are extremely notable.Immunopotentiator 1-4 group Swine PBMCs are after inactivation antigen stimulates, the IFN- of generation
γ is higher than inactivated vaccine group, statistical analysis significant difference；And immunopotentiator 1-3 groups are below 4 groups of immunopotentiator, statistical
It is not notable to analyse difference；As shown in figure 4,21 days after immune, the IFN-γ integral level that each group generates improves, immunopotentiator 1-4
Group is higher than inactivated vaccine group, statistical analysis significant difference；And immunopotentiator 1-3 groups are below 4 groups of immunopotentiator, statistical
It is not notable to analyse difference；Illustrated by ELISPOT result of the tests, 3 CTL epitope polypeptides, which are applied in combination, can enhance the thin of inactivated vaccine
Intracellular cytokine is horizontal.
To sum up, by the screening to predicting polypeptide, 3 CTL epitope polypeptides are obtained；By to CTL epitope polypeptides and inactivation
The combination application of vaccine, the immunopotentiator containing 3 CTL epitope polypeptides have significantly the immune efficacy of inactivated foot-and-mouth disease vaccine
Raising acts on, and can improve the cellular immune level and antibody level of vaccine.
The above is only the preferred embodiment of the present invention, it should be pointed out that：For the general of correlative technology field of the present invention
For logical technical staff, under the precondition for not departing from the principle of the invention, several improvement and modification can also be made, these change
Protection scope of the present invention is also should be regarded as into modification.
Sequence table
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<120>A kind of immunopotentiator and its application
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<213>Artificial sequence（Artificial sequence）
<400>2
Ala Leu Asp Asn Thr Thr Asn Pro Thr
1 5
<210> 3
<211>9
<212>PRT
<213>Artificial sequence（Artificial sequence）
<400>3
Leu Met Gln Thr Pro Ser His Thr Leu
1 5
<210> 4
<211> 10
<212>PRT
<213>Artificial sequence（Artificial sequence）
<400>4
Lys Thr Val Tyr Asn Gly Asn Cys Lys Tyr
1 5 10
<210> 5
<211> 10
<212>PRT
<213>Artificial sequence（Artificial sequence）
<400>5
Lys Arg Val Thr Glu Leu Leu Tyr Arg Met
1 5 10
<210> 6
<211>10
<212>PRT
<213>Artificial sequence（Artificial sequence）
<400>6
Lys Ala Leu Leu Arg Thr Ala Thr Tyr Tyr
1 5 10
<210> 7
<211> 9
<212> PRT
<213>Artificial sequence（Artificial sequence）
<400> 7
Lys Ala Pro Leu Thr Arg Leu Ala Leu
1 5
<210> 8
<211>9
<212>PRT
<213>Artificial sequence（Artificial sequence）
<400>8
Arg Arg His His Thr Asp Val Ser Phe
1 5
<210> 9
<211>9
<212>PRT
<213>Artificial sequence（Artificial sequence）
<400>9
Lys Ala Thr Arg Val Thr Glu Leu Leu
1 5
<210> 10
<211> 10
<212>PRT
<213>Artificial sequence（Artificial sequence）
<400>10
Lys Leu Ala Gln Lys Ala Ala Arg Pro Leu
1 5 10
Claims (10)
1. it is polypeptide in sequence table shown in sequence 1, more shown in sequence 2 in sequence table a kind of polypeptide for Immune-enhancing effect
Polypeptide in peptide or sequence table shown in sequence 3.
2. a kind of immunopotentiator, as the polypeptide and sequence shown in the polypeptide shown in sequence in sequence table 1, sequence 2 in sequence table
The peptide composition composition of one or both of polypeptide in table shown in sequence 3 any of the above combination.
3. immunopotentiator according to claim 2, which is characterized in that the sequence in the immunopotentiator is by sequence table
Two kinds of compositions in polypeptide in polypeptide and sequence table in polypeptide, sequence table shown in 1 shown in sequence 2 shown in sequence 3, they
Weight fraction ratio be 0.5-1：0.5-1；When the immunopotentiator as shown in sequence in sequence table 1 polypeptide, in sequence table
Polypeptide composition in polypeptide and sequence table shown in sequence 2 shown in sequence 3；Polypeptide, sequence in the sequence table shown in sequence 1
The ratio of weight and number of polypeptide in polypeptide or sequence table in table shown in sequence 2 shown in sequence 3 is 0.5-1：0.5-1：0.5-1.
4. immunopotentiator according to claim 3, which is characterized in that polypeptide, sequence in the sequence table shown in sequence 1
The ratio of weight and number of polypeptide in polypeptide and sequence table in list shown in sequence 2 shown in sequence 3 is 1：1：1.
5. application of the immunopotentiator in foot and mouth disease vaccine composition is prepared in claim 2-4 described in any one.
6. application of the polypeptide described in claim 1 in the immunopotentiator for preparing foot and mouth disease vaccine.
7. a kind of foot and mouth disease vaccine composition, the vaccine combination include：
1) immunopotentiator in claim 2-4 described in any one；
2) Schweineseuche antigen.
8. foot and mouth disease vaccine composition according to claim 7, which is characterized in that the foot and mouth disease vaccine composition
Further include veterinarily acceptable adjuvant.
9. foot and mouth disease vaccine composition according to claim 8, which is characterized in that the adjuvant is water-in-oil emulsion, water
One or more of bag oil emu, W/O/W emulsion；Preferably, the adjuvant is 206 adjuvants of Montanide ISA.
10. according to any one of the claim 7-9 foot and mouth disease vaccine compositions, which is characterized in that the Schweineseuche resists
Original is more selected from Schweineseuche inactivation antigen, swine foot-and-mouth disease virus sample particulate antigen, Schweineseuche subunit antigen or Schweineseuche
One or both of peptide antigen any of the above combines；Wherein, Schweineseuche inactivation antigen is Schweineseuche O-shaped or A type vaccines
Strain；Preferably, the pig FMDV inactivation antigens are selected from O/Mya98/XJ/2010 plants of pig, O/GX/09-7 plants, AF/72 plants, A/
The inactivation of viruses antigen of one or both of QH/09 plants any of the above combination is most preferably O/Mya98/XJ/2010 plants and goes out
Live virus antigen.
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| CN202010466664.2A CN111548389B (en) | 2018-01-03 | 2018-01-03 | Immunopotentiator and application thereof |
| CN201810003899.0A CN108117583B (en) | 2018-01-03 | 2018-01-03 | Immunopotentiator and application thereof |
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| CN201810003899.0A CN108117583B (en) | 2018-01-03 | 2018-01-03 | Immunopotentiator and application thereof |
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| CN202010466664.2A Division CN111548389B (en) | 2018-01-03 | 2018-01-03 | Immunopotentiator and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111925416A (en) * | 2020-08-26 | 2020-11-13 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum acquired immune response and application thereof |
| CN112300248A (en) * | 2020-08-26 | 2021-02-02 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof |
| CN113956329A (en) * | 2020-08-26 | 2022-01-21 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114751956A (en) * | 2020-08-26 | 2022-07-15 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof |
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| CN103724405B (en) * | 2013-12-30 | 2015-06-17 | 大连大学 | Type A foot-and-mouth disease CTL (cytotoxic T lymphocyte) epitope peptide and screening method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111925416A (en) * | 2020-08-26 | 2020-11-13 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum acquired immune response and application thereof |
| CN112300248A (en) * | 2020-08-26 | 2021-02-02 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof |
| CN112300248B (en) * | 2020-08-26 | 2021-12-03 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof |
| CN111925416B (en) * | 2020-08-26 | 2021-12-17 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum acquired immune response and application thereof |
| CN113956329A (en) * | 2020-08-26 | 2022-01-21 | 中国农业科学院兰州兽医研究所 | Polypeptide for promoting pig body to generate broad-spectrum immune response and application thereof |
| CN113956329B (en) * | 2020-08-26 | 2023-07-21 | 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111548389B (en) | 2021-11-26 |
| CN111548389A (en) | 2020-08-18 |
| CN108117583B (en) | 2020-06-09 |
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