CN112972669A - Animal experiment operation method for improving secretion of antibody of new corona vaccine to special population - Google Patents
Animal experiment operation method for improving secretion of antibody of new corona vaccine to special population Download PDFInfo
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- CN112972669A CN112972669A CN202110249246.2A CN202110249246A CN112972669A CN 112972669 A CN112972669 A CN 112972669A CN 202110249246 A CN202110249246 A CN 202110249246A CN 112972669 A CN112972669 A CN 112972669A
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- 229960005486 vaccine Drugs 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000002474 experimental method Methods 0.000 title claims abstract description 19
- 241001465754 Metazoa Species 0.000 title claims abstract description 16
- 230000028327 secretion Effects 0.000 title claims abstract description 15
- 230000006698 induction Effects 0.000 claims abstract description 29
- 241000700159 Rattus Species 0.000 claims abstract description 19
- 238000000338 in vitro Methods 0.000 claims abstract description 13
- 210000004027 cell Anatomy 0.000 claims description 20
- 238000002347 injection Methods 0.000 claims description 20
- 239000007924 injection Substances 0.000 claims description 20
- 241000711573 Coronaviridae Species 0.000 claims description 13
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 claims description 7
- 241000699670 Mus sp. Species 0.000 claims description 6
- 239000013642 negative control Substances 0.000 claims description 6
- 229940028617 conventional vaccine Drugs 0.000 claims description 4
- 230000036039 immunity Effects 0.000 claims description 4
- 238000002649 immunization Methods 0.000 claims description 4
- 230000003053 immunization Effects 0.000 claims description 4
- 229940031551 inactivated vaccine Drugs 0.000 claims description 3
- 238000010255 intramuscular injection Methods 0.000 claims description 2
- 239000007927 intramuscular injection Substances 0.000 claims description 2
- 238000010253 intravenous injection Methods 0.000 claims description 2
- 210000000689 upper leg Anatomy 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 10
- 241000700605 Viruses Species 0.000 abstract description 8
- 238000006243 chemical reaction Methods 0.000 abstract description 7
- 210000004698 lymphocyte Anatomy 0.000 abstract description 5
- 230000033228 biological regulation Effects 0.000 abstract description 3
- 238000001514 detection method Methods 0.000 abstract description 3
- 230000009191 jumping Effects 0.000 abstract description 2
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 208000002672 hepatitis B Diseases 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 230000004075 alteration Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000016784 immunoglobulin production Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 210000002955 secretory cell Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
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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
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
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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
- A61P31/14—Antivirals for RNA 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
- A61K2039/51—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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- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
- C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
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Abstract
The invention relates to the technical field of secretion regulation of virus specific antibodies, and discloses 30 white rats with weight of 250-300g and half female and half male, which are respectively layered and randomly divided into 3 groups according to the weight, and the 3 groups are respectively subjected to respective experimental operation and then subjected to sample collection and detection after being cultured. According to the animal experiment operation method for improving the antibody secretion of the new corona vaccine to special crowds, autologous lymphocytes are subjected to feedback after in vitro induction, data can be obtained in experiments to show that the antibody level of the in vitro induced lymphocyte autologous feedback is increased in a jumping mode, the experiment data can show that the operation method can overcome the slow reaction or non-reaction state of old people and weak and sick crowds to the vaccine, and the characteristic virus immune clearance effect is improved, so that the incidence rate of severe cases is reduced, and the operation method has a certain treatment effect on the severe cases.
Description
Technical Field
The invention relates to the technical field of virus specific antibody secretion regulation, in particular to an animal experiment operation method for improving the antibody secretion of a new corona vaccine to special people.
Background
With the development of molecular biology and immunology technologies, reasonable cell culture and induction technologies are utilized to direct the targeting functions of certain cells. We have succeeded in regulating the secretory cells of the antibody against hepatitis B by using the peripheral blood lymphocytes of the patient and using the positive healthy person of the antibody against hepatitis B as the regulation standard.
At present, the epidemic situation of the new coronavirus is not completely controlled, and a plurality of uncertain factors exist, and the problems of virus variation, poor vaccine reactivity of the old, uncertain treatment effect of severe patients and the like are the urgent difficulties to be researched and overcome.
This patent is based on animal experiment result, can successfully induce anti new coronavirus antibody production level in vitro, the suggestion improves serum antibody titer, overcomes old person and weak sick crowd and unresponsive or non-reactive state to the bacterin reaction, improves the immune clearance effect of characteristic virus to reduce the incidence of severe case, and, have certain treatment to severe case.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides an animal experiment operation method for improving the antibody secretion of a new corona vaccine to a special population, which has the advantages of improving the reactivity of an organism to the vaccine, improving the serum antibody titer, overcoming the slow response or non-response state of old people and weak and sick people to the vaccine, and improving the immune clearance effect of characteristic viruses, thereby reducing the incidence rate of severe cases, having a certain treatment effect on the severe cases and solving the problems in the background technology.
(II) technical scheme
In order to achieve the purpose, the invention provides the following technical scheme: 30 white rats with weight of 250-300g and half female rats and half male rats are respectively layered according to the weight and randomly divided into 3 groups, the 3 groups are respectively subjected to experimental operation, and sample collection and detection are performed after re-culture.
Preferably, the injection dosage of the new corona vaccine (vero inactivated vaccine) in the step 3 is 0.07 ug/Kg.
Preferably, the second day after the cell return transfusion in step 4 is performed with a conventional vaccine injection, the dosage and the frequency are the same as those of the antibody positive template group.
Preferably, the next day after the cell return transfusion in step 5, the conventional vaccine injection is performed, and the dosage and the frequency are the same as those of the antibody positive template group.
Preferably, the vaccine injection adopts intramuscular injection of thigh backs, the cell reinfusion adopts intravenous injection, the new coronavirus vaccine injection is performed 2-3 times at an interval of 2 weeks, the cell reinfusion is performed 1 time, the new coronavirus vaccine is 100 ul/time, and the cell reinfusion is performed 1 ml/time.
The invention aims to solve another technical problem of providing an animal experiment operation method for improving the secretion of the antibody of the new corona vaccine to special people, which comprises the following steps:
1) 30 white rats with the weight of 250-300g and half female rats and half male rats are respectively layered according to the weight and randomly divided into 3 groups;
2) the 3 groups are respectively 10 antibody positive template groups (normal immunity), 10 cell induction groups (autologous cell induction groups) and 10 negative control groups (non-induction groups);
3) carrying out conventional injection of a new corona vaccine on a white rat in the antibody positive template group until the antibody titer is greater than 1/1000 and taking the white rat as an induction template;
4) separating peripheral blood lymphocytes from the white mice in the cell induction group, performing autologous reinfusion after induction by taking the induction template in the step 3 as a standard, and performing conventional immunization injection of a new corona vaccine the next day after reinfusion;
5) separating peripheral blood lymphocytes from white mice of a negative control group, carrying out autologous reinfusion after in vitro non-induced culture, and carrying out conventional immune injection on a new corona vaccine the next day after reinfusion;
6) and (6) detecting the result.
(III) advantageous effects
Compared with the prior art, the invention provides an animal experiment operation method for improving the secretion of the new corona vaccine to the antibody of special people, which has the following beneficial effects:
according to the animal experiment operation method for improving the antibody secretion of the new corona vaccine to the special population, the antibody level of lymphocyte self-feedback induced in vitro by data obtained in the experiment is increased in a jumping manner by performing feedback after in vitro induction, and the experiment data can show that the operation method can overcome the slow reaction or non-reaction state of the old and the weak and sickly population to the vaccine, improve the immune clearance effect of characteristic viruses, so that the incidence rate of severe cases is reduced, and the operation method has a certain treatment effect on the severe cases.
Drawings
FIG. 1 is a line graph of experimental data of an animal experimental operation method for improving the secretion of antibodies of a new corona vaccine to a special population.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The first embodiment is as follows:
an animal experiment operation method for improving the secretion of antibodies of a new corona vaccine to a special population is characterized by comprising the following steps:
1) 30 white rats with the weight of 250-300g and half female rats and half male rats are respectively layered according to the weight and randomly divided into 3 groups;
2) the 3 groups are respectively 10 antibody positive template groups (normal immunity), 10 cell induction groups (autologous cell induction groups) and 10 negative control groups (non-induction groups);
3) carrying out conventional injection of a new crown vaccine on a white rat in the antibody positive template group, wherein the injection dose of the new crown vaccine (vero inactivated vaccine) is 0.07ug/Kg, and the antibody titer is greater than 1/1000 and is used as an induction template;
4) separating peripheral blood lymphocytes from the white mice in the cell induction group, performing autologous reinfusion after induction by taking the induction template in the step 3 as a standard, and performing routine immunization injection of a new corona vaccine the next day after reinfusion, wherein the dosage and the frequency are the same as those of the antibody positive template group;
5) separating peripheral blood lymphocytes from white mice of a negative control group, carrying out autologous reinfusion after in vitro non-induced culture, and carrying out conventional immunization injection of a new corona vaccine the next day after reinfusion, wherein the dosage and the times are the same as those of an antibody positive template group;
6) and (3) detecting results, collecting about 0.5mL of blood by using an inert separation gel accelerating tube before administration and two weeks after each administration of 3 groups of white rats, putting the collected blood into a sample transport box, and sending the blood to a clinical examination department for detection within 2 h.
Experimental example:
and (3) detecting the titer of the inactivated protein of the new coronavirus in the supernatant on the fifth day of culture in a culture medium containing 2ng/ml of the new coronavirus vaccine, wherein the in-vitro induction group generates a negative result after being diluted by 1/8, and the non-in-vitro induction group is negative after being converted into 1/64, which is shown in table 1. On the 10 th day of culture with 4ng/ml culture medium containing new coronavirus vaccine, the titer of the inactivated protein of the new coronavirus in the supernatant is detected, a negative hole appears when the in vitro induction group is diluted by 1/32 times, the protein turns negative completely when the protein turns negative at 1/64, and the protein does not turn negative at the dilution in the non-in vitro induction group, which is shown in table 2.
TABLE 1 viral antigen titer in supernatants of fifth day of culture containing the New coronavirus vaccine 2ng/ml
TABLE 2 viral antigen titer in supernatants at day 10 of culture containing the novel coronavirus vaccine 4ng/ml
And (4) judging the standard: antibody levels.
The invention has the beneficial effects that: this improve animal experiment operation method that new coronary vaccine secretly to special crowd's antibody, autologous lymphocyte carries out the feedback through inducing in vitro, the data that can draw shows in the experiment, the antibody level of the autologous feedback of the lymphocyte of external induction is the jump rising, this experimental data can show that this operation method can overcome old person and weak sick crowd to the vaccine reaction slow or the non-reaction state, improve characteristic virus immunity clearance effect, thereby reduce the incidence of severe case, and, have certain therapeutic action to severe case.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (5)
1. An animal experiment operation method for improving the secretion of the antibody of the new corona vaccine to special people is characterized by comprising an animal experiment and the following steps:
1) 30 white rats with the weight of 250-300g and half female rats and half male rats are respectively layered according to the weight and randomly divided into 3 groups;
2) the 3 groups are respectively 10 antibody positive template groups (normal immunity), 10 cell induction groups (autologous cell induction groups) and 10 negative control groups (non-induction groups);
3) carrying out conventional injection of a new corona vaccine on a white rat in the antibody positive template group until the antibody titer is greater than 1/1000 and taking the white rat as an induction template;
4) separating peripheral blood lymphocytes from the white mice in the cell induction group, performing autologous reinfusion after induction by taking the induction template in the step 3 as a standard, and performing conventional immunization injection of a new corona vaccine the next day after reinfusion;
5) separating peripheral blood lymphocytes from white mice of a negative control group, carrying out autologous reinfusion after in vitro non-induced culture, and carrying out conventional immune injection on a new corona vaccine the next day after reinfusion;
6) and (6) detecting the result.
2. The animal experimental operation method for improving the antibody secretion of the new crown vaccine to the special population according to claim 1, wherein the injection dosage of the new crown vaccine (vero inactivated vaccine) in the step 3 is 0.07 ug/Kg.
3. The method of claim 1, wherein the second day after cell transfusion in step 4 is performed with conventional vaccine injection at the same dosage and frequency as the antibody positive template set.
4. The method of claim 1, wherein the second day after cell transfusion in step 5 is performed with conventional vaccine injection at the same dosage and frequency as the antibody positive template set.
5. The animal experiment operation method for improving the antibody secretion of the new coronavirus vaccine to the special population as claimed in claim 1, wherein the vaccine injection is intramuscular injection of thigh back, the cell reinfusion is intravenous injection, the new coronavirus vaccine injection is performed 2-3 times at an interval of 2 weeks, the cell reinfusion is 1 time, the new coronavirus vaccine is 100 ul/time, and the cell reinfusion is 1 ml/time.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002034119A2 (en) * | 2000-10-27 | 2002-05-02 | Immuno-Rx, Inc. | Vaccine immunotherapy for immune suppressed patients |
| CN1480215A (en) * | 2003-07-07 | 2004-03-10 | 叶新新 | Compound antibody vaccine of SARS virus antigen as well as its model of experimental animal and method |
| CN105132371A (en) * | 2015-07-24 | 2015-12-09 | 北京鼎成肽源生物技术有限公司 | Method for in vitro presenting and activating DC cells and application of method |
| CN111569058A (en) * | 2020-06-18 | 2020-08-25 | 武汉生物制品研究所有限责任公司 | SARS-CoV-2 inactivated vaccine and its preparation method |
-
2021
- 2021-03-08 CN CN202110249246.2A patent/CN112972669A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002034119A2 (en) * | 2000-10-27 | 2002-05-02 | Immuno-Rx, Inc. | Vaccine immunotherapy for immune suppressed patients |
| CN1480215A (en) * | 2003-07-07 | 2004-03-10 | 叶新新 | Compound antibody vaccine of SARS virus antigen as well as its model of experimental animal and method |
| CN105132371A (en) * | 2015-07-24 | 2015-12-09 | 北京鼎成肽源生物技术有限公司 | Method for in vitro presenting and activating DC cells and application of method |
| CN111569058A (en) * | 2020-06-18 | 2020-08-25 | 武汉生物制品研究所有限责任公司 | SARS-CoV-2 inactivated vaccine and its preparation method |
Non-Patent Citations (4)
| Title |
|---|
| ADAM D. COHEN 等: "Autologous lymphocyte infusion supports tumor antigen vaccine-induced immunity in autologous stem cell transplant for multiple myeloma", 《AMERICAN ASSOCIATION FOR CANCER RESEARCH》, pages 1 - 39 * |
| BRIAN HANLEY 等: "Convalescent donor SARS-COV-2-specific cytotoxic T lymphocyte infusion as a possible treatment option for COVID-19 patients with severe disease has not received enough attention till date", 《BRITISH JOURNAL OF HAEMATOLOGY》, vol. 189, pages 1050 * |
| 江梦颖 等: "新型冠状病毒肺炎与血浆输注治疗", 《镇江高专学报》, vol. 33, no. 3, pages 109 - 112 * |
| 蔡勇 等: "体外诱导的同种反应性淋巴细胞输注对 大鼠皮肤移植存活时间的影响", 《实验动物科学》, vol. 25, no. 2, pages 16 - 19 * |
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