CN106177941A - A kind of adenovirus utilizing expression of GM CSF improves method and the test kit of newcastle inactivated vaccine immune effect - Google Patents
A kind of adenovirus utilizing expression of GM CSF improves method and the test kit of newcastle inactivated vaccine immune effect Download PDFInfo
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- CN106177941A CN106177941A CN201610648941.5A CN201610648941A CN106177941A CN 106177941 A CN106177941 A CN 106177941A CN 201610648941 A CN201610648941 A CN 201610648941A CN 106177941 A CN106177941 A CN 106177941A
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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
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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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5252—Virus inactivated (killed)
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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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- 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
Abstract
The present invention relates to microorganism and biological technical field, particularly relate to a kind of method and test kit utilizing the adenovirus expressing granulocyte macrophage colony stimulating factor to improve newcastle inactivated vaccine immune effect.The inventive method includes recombinant adenovirus and the newcastle inactivated vaccine of expression of GM CSF are uniformly mixed so as to obtain vaccine immunity adjuvant mixture, then utilizes above-mentioned vaccine immunity adjuvant mixture that birds is carried out immunity inoculation.Present invention additionally comprises a kind of expression of GM CSF recombinant adenovirus constitute immunological adjuvant and the recombinant adenovirus of a kind of expression of GM CSF and newcastle inactivated vaccine composition test kit, said method and test kit can be used for poultry cultivation industry.
Description
Technical field
The present invention relates to microorganism and biological technical field, particularly relate to a kind of adenovirus utilizing expression of GM-CSF and improve
The method of newcastle inactivated vaccine immune effect and test kit.
Background technology
Newcastle (Newcastle Disease, ND) is the one being caused the birdss such as chicken, turkey, Columba livia by Avian pneumo-encephalitis virus
Acute, high degree in contact infectiousness epidemic disease, mainly by causing birds septicemia to cause it dead, is that fowl poultry husbandry is supported in harm
One of three big infectious disease, in Asia, a lot of countries and regions cause huge harm to this epidemic disease for many years.Reply is new at present
City epidemic disease most effective way injects newcastle inactivated vaccine exactly, can effectively prevent generation and the propagation of birds newcastle epidemic disease.
Immunological adjuvant, also known as nonspecific immunity proliferant agent, is a kind of itself not have an antigenicity, but synantigen is together or in advance
Can effectively strengthen immunogenicity in being first expelled to body or change the nonspecific immunity strengthening agent of immunoreation type.Immunity assistant
Agent kind is a lot, and conventional adjuvant can be divided into 4 classes: inorganic adjuvant, such as aluminium hydroxide, calcium phosphate, Alumen etc.；Organic adjuvant, micro-
Biological and product such as mycobacteria (tubercule bacillus, bacillus calmette-guerin vaccine), bacillus pumilis, escherichia coli, bordetella pertussis, endotoxin,
Bacterial extract (muramyldipeptide) etc.；Synthetic adjuvant, as synthetic double stranded polynucleotide (double-strand polyadenylic acid,
Uridylic acid), levamisole, inosine pranobex, liposome, surfactant etc.；Oil preparation, such as Freund adjuvant (Freund
Adjuvant), adjuvant 65, mineral oil, vegetable oil etc..Freund adjuvant is the most frequently used in laboratory animal at present, again may be used
Being divided into incomplete Freund's adjuvant and Freund's complete adjuvant two kinds, Freund's incomplete adjuvant can be used for inoculation, the immune intensity of Freund's complete adjuvant
More than Freund's incomplete adjuvant, it is mainly used in zoopery, is unsuitable for the mankind and uses, and the most often can occur after animal multiple injection
Adjuvant disease.
Granulocyte-macrophage colony stimutaing factor (granulocyte macrophagecolony stimulating
Factor, GM-CSF) it is a kind of multi-functional somatomedin, research shows, GM-CSF can stimulate proliferation of hematopoietic progenitors, differentiation
Move to outer planet with maturation and from bone marrow, various kinds of cell differentiation, propagation can be induced.Granulocyte colony stimulating factor is also commonly used as
Immunological adjuvant, its immunoenhancement result much studied confirmation, when it uses as the immunological adjuvant of newcastle inactivated vaccine,
Can increase the volume of antigen in being inoculated into poultry body, easily be absorbed by antigen presenting cell (APC), extend antigen depositing in vivo
Stay the phase, increase the opportunity contacted with immunocyte, the beneficially propagation of immune stimulatory cell.
But, generally using GM-CSF as immunological adjuvant use time, eucaryon or prokaryotic expression system can be used as submission
Carrier, its defect is often cannot well to play the effect of immunological adjuvant, greatly because the half-life of GM-CSF is short
Have impact on its immunoenhancement result.The present invention, through repeatedly attempting, have finally chosen the adenovirus delivery vehicle as GM-CSF,
Make full use of adenovirus good stability, the advantage of appeal can be preserved for a long time, overcome GM-CSF half-life short drawback, carry
Rise the immunoenhancement result of GM-CSF, further increase GM-CSF and strengthen the immunity of newcastle inactivated vaccine as immunological adjuvant
The use effective percentage of activity.Provided by the present invention this by improving immunological adjuvant delivery vehicle and then improving this adjuvant to newly
Also someone reports the method for city epidemic disease inactivated vaccine immunoenhancement result at present.The method of the present invention and test kit thereof can extensively be fitted
For bird immunity industry, the popularization and application of the technology of the present invention will have good market prospect and produce considerable economy and
Social benefit.
Summary of the invention
Solve the technical problem that
The present invention needs the problem solved to be: it is an object of the invention to overcome using GM-CSF as immunological adjuvant use time,
Use eucaryon or prokaryotic expression system cannot be played very well because the half-life of GM-CSF is short as what delivery vehicle caused
The effect of immunological adjuvant, and the technical problem of its immunoenhancement result of extreme influence.
Technical scheme
It is desirable to provide a kind of adenovirus utilizing expression of GM-CSF improves newcastle inactivated vaccine immune effect
Method, using solve to presently, there are by GM-CSF as immunological adjuvant use time, use eucaryon or prokaryotic expression system as passing
That caused in carrier because half-life of GM-CSF is short and cannot play the effect of immunological adjuvant very well, and extreme influence its exempt from
The technical barrier of epidemic disease reinforced effects.
The present invention improves the method for newcastle inactivated vaccine immune effect, comprises the following steps:
(1) recombinant adenovirus and the mixing of newcastle inactivated vaccine of granulocyte-macrophage colony stimutaing factor will be expressed
Obtain vaccine immunity adjuvant mixture；
(2) utilize above-mentioned vaccine immunity adjuvant mixture that birds is carried out immunity inoculation.
As a kind of preferred version, the restructuring expressing granulocyte-macrophage colony stimutaing factor described in the method
Adenovirus is the people five type recombinant adenovirus expressing granulocyte-macrophage colony stimutaing factor.
Further, utilize said method birds to be carried out during immunity inoculation and use chest muscle injection system.
Additionally, present invention also offers a kind of immunological adjuvant for improving newcastle inactivated vaccine immune effect, this is exempted from
Epidemic disease adjuvant is the recombinant adenovirus expressing granulocyte-macrophage colony stimutaing factor.
As a kind of preferred version, this immunological adjuvant is people five type expressing granulocyte-macrophage colony stimutaing factor
Recombinant adenovirus.
A kind of make according to the method for above-mentioned raising newcastle inactivated vaccine immune effect additionally, present invention also offers
Test kit, this test kit includes recombinant adenovirus and the newcastle inactivation expressing granulocyte-macrophage colony stimutaing factor
Vaccine.
As a kind of preferred version, this test kit includes the people five expressing granulocyte-macrophage colony stimutaing factor
Type recombinant adenovirus and newcastle inactivated vaccine.
Further, during test kit of the present invention can be widely applied to poultry cultivation industry.
Beneficial effect
Provided by the present invention this by improving immunological adjuvant delivery vehicle and then improving this adjuvant to newcastle inactivation
The method of vaccine immunity reinforced effects overcomes the major defect that prior art exists, and is the immunostimulant improving immunological adjuvant
Effect provides simple and easy to do technical scheme.The present invention, through repeatedly attempting, have finally chosen adenovirus as GM-CSF's
Delivery vehicle, makes full use of adenovirus good stability, can preserve the advantage of appeal for a long time, overcome the GM-CSF half-life short
Drawback, promote GM-CSF immunoenhancement result, further increase GM-CSF as immunological adjuvant strengthen newcastle inactivation
The use effective percentage of vaccine immunity activity.The method of the present invention and test kit thereof are widely portable to bird immunity industry, this
The popularization and application of bright technology will have good market prospect and produce considerable economy and social benefit.
Accompanying drawing explanation
Fig. 1 is newcastle epidemic disease antibody change after immunity.Separate serum blood sampling in 7,14,21 and 28 days, use hemagglutination inhibition test
(HI) detection antibody titer, compares antibody variability by the log2 value of HI, when p≤0.05, significant difference (*).
Fig. 2 is immunity related molecular expression change after booster immunization.Internal reference is done with 28s, all groups and without counteracting toxic substances matched group
Relatively, when p≤0.05, significant difference (*).
Fig. 3 is clinical symptoms evaluation and survival rate statistics after counteracting toxic substances.Daily clinical index: Clinical scores refers to
Number；Percent survival: survival rate；Days post infection (DPI): DAI.
Fig. 4 is for tissue and observes Pathologic Observation.A: glandular stomach microscope pathological changes (H.E dyes, 10 × 40)；B: lung is micro-
Mirror pathological changes (he dyes, 10 × 40)；C: duodenum pathological changes is observed.
Fig. 5 is tissue virus load detection.5d gathers the heart (heart) of infected chicken, lungs (lung), stomach after infection
(stomach), duodenum (Duodenum) and fabricius bursa (Bursa fabricii), drip by real-time quantitative PCR detection virus
Degree, when p≤0.05, significant difference (*).
Detailed description of the invention
Below by way of specific instantiation, embodiments of the present invention being described, those skilled in the art can be by this specification
Disclosed content understands other advantages and effect of the present invention easily.The present invention can also be by the most different concrete realities
The mode of executing is carried out or applies, the every details in this specification can also based on different viewpoints and application, without departing from
Various modification or change is carried out under the spirit of the present invention.
Before further describing the specific embodiment of the invention, it should be appreciated that protection scope of the present invention is not limited to down
State specific specific embodiments；It is also understood that the term used in the embodiment of the present invention is specific concrete in order to describe
Embodiment rather than in order to limit the scope of the invention；In description of the invention and claims, unless in literary composition
Additionally explicitly pointing out, singulative " ", " one " and " this " include plural form.
When embodiment provides numerical range, it should be appreciated that unless the present invention is otherwise noted, two ends of each numerical range
Between point and two end points, any one numerical value all can be selected for.Unless otherwise defined, in the present invention use all technology and
The same meaning that scientific terminology and those skilled in the art of the present technique are generally understood that.Except in embodiment use concrete grammar, equipment,
Outside material, according to those skilled in the art's grasp to prior art and the record of the present invention, it is also possible to use and this
Any method, equipment and the material of the prior art that the method described in inventive embodiments, equipment, material are similar or equivalent comes real
The existing present invention.
Embodiment 1
The present invention utilizes adenovirus and the newcastle disease virus inactivated vaccine co-immunization expressing chicken GM-CSF, it is possible to notable
Promote the body fluid of newcastle inactivated vaccine, cellular immune level, strengthen the immunoprotection effect that Virulent Newcastle Disease Virus (F48E9) is infected
Really.
1. experiment material
Virus: wild type human five type adenovirus (wAd), the recombinant adenovirus (rAd-GM-CSF) of expression of GM-CSF, new city
Epidemic disease inactivated vaccine (InV).
Laboratory animal: chickling 126.
2. test method
2.1 animal immunes and challenge test
Test chicken is divided into 6 groups, and often group 21, processes according to design shown in table 1, respectively at 7 ages in days
Carrying out just exempting from, 21 ages in days carry out two and exempt from.Immunity uses chest muscle injection, and the group of adenovirus immunity simultaneously, in advance by adenopathy
Poison and inactivated vaccine mix.
Table 1 animal experiment design
Two exempt from the rear Virulent Newcastle Disease Virus F48E9 strain containing 108TCID50 with 0.1ml in 2 weeks carries out counteracting toxic substances, evaluates immunity and protects
Protect effect.
2.2 antibody test
Collecting serum antibody 0,14,21,28 and 35d, the method recommended with OIE carries out blood clotting (HA) and blood clotting suppression examination
Test (HI), detect serum antibody titer of ND.
Gathering healthy cock erythrocyte, the erythrocyte of preparation 1%, normal saline, as diluent, dilutes 4 unit antigens
(using Avian pneumo-encephalitis virus la Sota strain as standard antigen), evaluates serum antibody titer by the log2 value of HI.
2.3 cellular immunization correlation molecule detections
Study On Cellular Immune is evaluated with splenocyte IFN-а, IFN-β, IFN-γ, and the expression of IL-4.At 22d
(24h after immunity), puts to death 5 chickens, gathers spleen, extracts RNA and detects IFN-а, IFN-β, IFN-γ, and the expression of IL-4.With
The method of relative quantification evaluates the change of the expression of several immunity correlation factor, and every part of sample does 3 repetitions, selects 28s to make
For internal reference (Δ Ct=Ct target Ct internal reference), then compare (2-Δ Δ CT, Δ Δ CT=Δ Ct process group-Δ Ct with non-group of exempting from
Matched group), weigh promising result by the multiple being raised and lowered.
2.4 counteracting toxic substances protected effect detections
2.4.1 clinicing symptom observation
The clinical manifestation (normal 0, depressed 1, paralysis 2, dead 3) of chicken is evaluated by the mode of marking.Observe 14 days after infection,
Record marking result, evaluates the incidence of infected chicken, records mortality rate simultaneously with score averages.
2.4.2 pathological study
5d after infection, puts to death 3 chickens, takes lungs and glandular stomach makes pathological section, observes what newcastle infection caused
Histopathologic change difference.
2.4.3 tissue viral level detection
5d after infection, puts to death 5 chickens, detects in the heart, lung, Stomach duodenum and fabricius bursa with real-time quantitative PCR
Virus load, primer is for the M gene of NDV, and sequence is as shown in table 2.
Primer used by table 2 Real_time quantitative detection
Detection uses the SYBR Green of Vazyme (Cat.No Q111-02, Vazyme Biotech co., ltd) quantitative
Detection kit, instrument uses sky, Xi'an grand four-way real-time PCR.
2.4.4 toxin expelling detection
3,5,7d collection trachea and cloacal swabs, be placed in 1 milliliter of PBS containing antibiotic, through place after infection
Inoculate SPF chicken embryo after reason, by viral level in hemagglutination test detection Embryo Gallus domesticus urine, analyze toxin expelling situation.
3. result of the test
3.1 antibody titer of ND
As it is shown in figure 1, two exempt from rear immunological sterilization vaccine and the group (InV+GM) of expression of GM-CSF recombinant adenovirus simultaneously, obtain
Obtaining the highest antibody titer, mean titre is about 12 (log2)；Group (InV) antibody titer of immunological sterilization vaccine is about 10
(log2), significant difference, other group antibody titer is about 6 (log2).The above results shows the gland of Immune expression GM-CSF simultaneously
Virus can significantly improve the antibody horizontal of immunity chicken.
3.2 cell immunity of spleen correlation factor expressions
As in figure 2 it is shown, two exempt from rear 24h, the chicken spleen cell IFN-а of InV+GM group, IFN-β, IFN-γ, and the expression of IL-4
Amount is significantly higher than other group, significant difference (p < 0.05) compared with InV group.Show the adenovirus of Immune expression GM-CSF simultaneously
The transcriptional level of immunity related molecular can be significantly improved.
3.3 clinical symptoms evaluation and survival rates
InV+GM group Clinical scores is about 0.5, and InV group Clinical scores is about 1.5.
Being vaccinated with the group of wAd, 7d after infection is the most dead for chicken；The group (CC) of nonimmune counteracting toxic substances comparison, chicken is infecting
Rear 8d is the most dead；It is vaccinated with expression granulocyte colony stimulating factor, but does not inoculate the group (rAd-of newcastle inactivated vaccine
GM) 10d is the most dead after infection.
As it is shown on figure 3, InV group survival rate is 46% (7/15), the survival rate of InV+GM group is 86% (13/15).Show
The adenovirus of Immune expression GM-CSF can reduce the morbidity order of severity and the mortality rate that strong poison infection causes simultaneously.
3.4 observe and histopathologic change
As shown in Figure 4,5d after infection, cuts open 3 chickens of inspection, evaluates pathological changes situation, find CC, wAd and rAd-GM group
Chicken on multiple histoorgans it can be seen that the most hemorrhage.InV group can be seen slight hemorrhage at intestinal serosal surface, and
InV+GM group then can not see bleeding.
Histopathological examination finds, the most hemorrhage and inflammatory seen from matched group CC, wAd and rAd-GM lungs and glandular stomach
The infiltration of cell；The visible glandular stomach pipe Mild edema of InV group and lungs hemorrhage, and the chicken exempting from InV+GM group is visible on glandular stomach pipe
Slighter edema, lungs then visible pathological change.Show that the adenovirus of Immune expression GM-CSF simultaneously can lower strong poison
What infection caused observes histopathologic change and microscopic structure pathological change.
3.5 tissue viral level and toxin expellings
As it is shown in figure 5, matched group tissue viral level is about 103.5～104.2 copies/100ul, the chicken tissues of InV group
Viral level is about 102.6～102.9 copies/100ul, and the chicken tissues viral level about 102.1～102.4 of InV+GM group is copied
Shellfish/100ul.Show that the adenovirus of Immune expression GM-CSF simultaneously can significantly reduce tissue virus load.
It can be seen that toxin expelling detection finds that matched group is after infection 3,5 and 7d all the quantity of toxin expelling chicken after table 3 infects
There is serious toxin expelling phenomenon, and InV group after infection 3,5 and 7d toxin expelling rate be respectively 40%, 20% and 20%；InV+GM
Group toxin expelling rate is respectively 0%, 20% and 0%.Show that the adenovirus of Immune expression GM-CSF simultaneously can significantly reduce toxin expelling rate.
The quantity of toxin expelling chicken after table 3 infection
Note * DAI
。
Claims (8)
1. the method improving newcastle inactivated vaccine immune effect, comprises the following steps:
(1) recombinant adenovirus and newcastle inactivated vaccine of expressing granulocyte-macrophage colony stimutaing factor are uniformly mixed so as to obtain
Vaccine immunity adjuvant mixture；
(2) utilize above-mentioned vaccine immunity adjuvant mixture that birds is carried out immunity inoculation.
2. the method improving newcastle inactivated vaccine immune effect as claimed in claim 1, it is characterised in that: described expression grain
The recombinant adenovirus of granulocytemacrophage colony stimulating factor is the people five expressing granulocyte-macrophage colony stimutaing factor
Type recombinant adenovirus.
3. the method improving newcastle inactivated vaccine immune effect as claimed in claim 1 or 2, it is characterised in that: utilize institute
State vaccine immunity adjuvant mixture to carry out birds using chest muscle injection system during immunity inoculation.
4. the immunological adjuvant being used for improving newcastle inactivated vaccine immune effect, it is characterised in that: described immunological adjuvant is
Express the recombinant adenovirus of granulocyte-macrophage colony stimutaing factor.
5. the immunological adjuvant for improving newcastle inactivated vaccine immune effect as claimed in claim 4, it is characterised in that: institute
Stating immunological adjuvant is the people five type recombinant adenovirus expressing granulocyte-macrophage colony stimutaing factor.
6. the test kit that the method for raising newcastle inactivated vaccine immune effect according to claim 1 is made, institute
State test kit and include expressing recombinant adenovirus and the newcastle inactivated vaccine of granulocyte-macrophage colony stimutaing factor.
7. test kit as claimed in claim 6, it is characterised in that: described expression granulocyte-macrophage colony stimutaing factor
Recombinant adenovirus be express granulocyte-macrophage colony stimutaing factor people five type recombinant adenovirus.
The application in poultry cultivation industry of the test kit the most as claimed in claims 6 or 7.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1258738A (en) * | 1998-12-31 | 2000-07-05 | 上海华晨生物技术研究所 | Recombined adenovirus expressing the stimulating factor of human granulocyte-macrophage colony and its preparation and use |
| CN1381562A (en) * | 2001-04-13 | 2002-11-27 | 上海华康生物技术有限公司 | Antigen-sensitive human GM-CSF gene modified human dendritic cell and its preparing process and usage |
| CN1381582A (en) * | 2001-04-13 | 2002-11-27 | 上海华康生物技术有限公司 | Targetting high-expression recombinant adenovirus of human granulocyte-macrophage colony stimulating factor and its preparing process and usage |
| CN101423823A (en) * | 2008-10-29 | 2009-05-06 | 南京农业大学 | Porcine reproductive and respiratory syndrome recombinant adenovirus rAd-GF35 |
| CN103834620A (en) * | 2013-07-19 | 2014-06-04 | 北京中联康生物科技有限公司 | Newcastle disease virus, newcastle disease inactivated vaccine and preparation method of newcastle disease inactivated vaccine |
| CN104164410A (en) * | 2014-08-07 | 2014-11-26 | 哈尔滨博翱生物医药技术开发有限公司 | Newcastle disease virus strain and application thereof in preparation of Newcastle disease vaccine |
-
2016
- 2016-08-09 CN CN201610648941.5A patent/CN106177941A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1258738A (en) * | 1998-12-31 | 2000-07-05 | 上海华晨生物技术研究所 | Recombined adenovirus expressing the stimulating factor of human granulocyte-macrophage colony and its preparation and use |
| CN1381562A (en) * | 2001-04-13 | 2002-11-27 | 上海华康生物技术有限公司 | Antigen-sensitive human GM-CSF gene modified human dendritic cell and its preparing process and usage |
| CN1381582A (en) * | 2001-04-13 | 2002-11-27 | 上海华康生物技术有限公司 | Targetting high-expression recombinant adenovirus of human granulocyte-macrophage colony stimulating factor and its preparing process and usage |
| CN101423823A (en) * | 2008-10-29 | 2009-05-06 | 南京农业大学 | Porcine reproductive and respiratory syndrome recombinant adenovirus rAd-GF35 |
| CN103834620A (en) * | 2013-07-19 | 2014-06-04 | 北京中联康生物科技有限公司 | Newcastle disease virus, newcastle disease inactivated vaccine and preparation method of newcastle disease inactivated vaccine |
| CN104164410A (en) * | 2014-08-07 | 2014-11-26 | 哈尔滨博翱生物医药技术开发有限公司 | Newcastle disease virus strain and application thereof in preparation of Newcastle disease vaccine |
Non-Patent Citations (1)
| Title |
|---|
| 王卉等: "鞭毛蛋白及鸡GM-CSF重组新城疫病毒对鸡的免疫增强作用", 《中国预防兽医学报》 * |
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