Method for prophylaxis of radiation damage

FIELD: medicine, radiation medicine.

SUBSTANCE: method involves subcutaneous administration of anti-influenza vaccine "Grippol" in the effective dose. Invention provides enhancing effectiveness of body to effect of ionizing radiation. Invention can be used for prophylaxis of radiation damage.

EFFECT: improved method of prophylaxis.

5 tbl, 2 ex

 

The invention relates to medicine, namely to methods for prevention of acute radiation sickness.

It is known that various substances of microbial origin have increased radioresistance of the organism [1]. Radioprotective effect have many vaccines, including viral. A positive effect was obtained with the use of smallpox vaccine experiments on rabbits [2]. Despite the pronounced radioprotective effect of smallpox vaccine, it cannot be used as a method of prevention of radiation injuries, since the eradication of smallpox in all countries eliminated smallpox vaccine from the number of drugs used in medical practice.

Much more promising as a drug dual-purpose (anti-infective and radioprotective) is the flu vaccine because the flu every year suffer hundreds of millions of people. For immunization against influenza until recently used liquid inactivated vaccine Omutninsk chemical plant of Kirov region. The vaccine contained a flu-like virus antigens of type A1 and A2, peeled chromatographic method. This vaccine was examined for its ability to increase the radioresistance of the organism [3]. It has been shown that prophylactic subcutaneous administration of the vaccine to mice (02 ml) or hamsters (0.2 ml) at various times before irradiation (1-30 day) by 10-40% enhances survival of irradiated and protected animals compared with unvaccinated [3]. However, the above-mentioned vaccine currently discontinued due to its low specific immunogenic properties and in medical practice is not applied.

Now for the prevention of influenza used vaccine "Grippol" [4]. It is different from the liquid inactivated vaccine production Omutninsk chemical plant that is composed of a water-soluble high-molecular immune - N-oxidized derivative of poly-1,4-ethyleneimine (Polyoxidonium), and also contains surface glycoproteins (hemagglutinin and neuraminidase), isolated from purified influenza viruses type a and b [5]. One immunizing dose of the vaccine "Grippol" for subcutaneous injection (0.5 ml) contains, according to the instructions for use, 5 µg of hemagglutinin of influenza virus strains of type a (subtype H1N1and H3N2), 11 μg of hemagglutinin of influenza virus strains of type b and 500 µg polyoxidonium. Vaccine "Grippol" free from impurities Averianova origin and is a highly effective and safe drug. It was developed by the Institute of immunology FMBA. Vaccine "Grippol" is widely used as a means for the prevention of influenza [5]. Now she has immunized more than 40 million people. Monitoring results vacci the purpose of the population indicated the safety, good tolerability and high prophylactic efficacy of the drug. Therefore, the vaccine "Grippol" approved for widespread use in epidemiological practice for prophylactic immunization of children starting from the age of 6 months, and adults [6]. Its use is indicated for populations at high risk of influenza.

Hitherto remained unexplored question about the radioprotective efficacy of the vaccine "Grippol", which, of course, is relevant and has not only scientific, but also practical interest. In case of positive results, the vaccine "Grippol" can be recommended as the drug of double action: anthropologi and radioprotective. It can be used for immunization of employees of the nuclear industry, the soldiers and officers of the Russian Army, the staff of the Ministry and other groups at risk of radiation exposure.

The aim of the invention is to develop a new method of prevention of radiation injuries from the vaccine "Grippol", widely used for vaccination against influenza.

As a result, we found that the introduction into the organism influenza vaccine "Grippol" subcutaneously before irradiation significantly increases the body's resistance to ionizing radiation. The method is simple and easy to use.

For achieving the set of goals has been studied the radioprotective effects of the vaccine "Grippol" on mice hybrids (NEA× C57B1)F1the males, weighing 18-23 g, in experimental acute radiation sickness, which is caused by irradiation of mice by gamma-quanta137Cs installation IGUR. The dose rate of 1.2 to 2.0 rad/sec. Monitoring of dose rate and its spatial distribution was performed using thermoluminescent dosimeter X-AND using thermoluminescence aluminophosphate glass IP-7 error ±10%. As the model used a single uniform radiative forcing that causes bone marrow form of acute radiation disease of varying severity. Were used doses with different biological activity: 6,0 (SD10/30); and 7.5 (SD10-70/30); and 8.5 (SD100/30) Gr. Observation of animals began immediately and lasted for 30 days after irradiation. Irradiation of mice was performed in a special plastic containers, divided into individual cells. The vaccine "Grippol" was administered to mice as a radioprotective funds once subcutaneously in doses of 0.5, 0.2 and 0.05 ml for 1, 7 or 14 days before irradiation. Along with groups protected irradiated mice were generated corresponding group irradiated control animals, which instead vaccine "Grippol" was injected with saline. The randomization of the control and experimental groups was carried out on the body weight of each group was 10-12 mice. Irradiation at the indicated doses led to development in mice typical bone marrow form of acute radiation sickness, which was manifested by decreased motor activity and food motivation. The mouse was untidy, the body weight they dropped. In the bone marrow and in peripheral blood decreased number of cells. The death of the irradiated animals took place in the midst of acute radiation sickness (7th - 20th day after irradiation). About the effectiveness of the vaccine "Grippol" judged on the basis of accounting as the lifespan of mice with a dose of 8.5 Gy; survival rates and body weight after irradiation at a dose of 7.5 G; the content of cells in the bone marrow and peripheral blood at a dose of 6.0 Gy. The obtained data were processed by standard methods of variation statistics using the computer program "patch BIOSTAT" [7].

In the course of the research found in the vaccine "Grippol" the new property, which is manifested in the increase of resistance of immunized mice to ionizing radiation.

To determine the optimal radioprotective doses of the vaccine "Grippol" mice were immunized by it in the amount of 0,5; 0,2 or 0.05 ml Dose was 7.5 Gr. However, in this experiment, the survival rate in the control group of animals was high and amounted to 91.6%. Therefore, to determine the optimal radioprotective doses of the vaccine used is ovali indicators of body weight of the mice (table 1).

Table 1

Change of body weight in mice protected by the vaccine "Grippol"entered in various amounts up to 7 days before irradiation by gamma-quanta137Cs at a dose of 7.5 Gy (M±m, g)
The vaccine dose, mlThe time of examination after exposure, day.
07101520
0,523,6±0,320,5±0,920,8±0,921,5±0,722,5±0,5
0,223,8±0,122,0±0,322,0±0,322,4±0,423,4±0,5
0,0524,1±0,221,9±0,421,1±0,719,7±0,921,7±1,0
Control exposure23,1±0,621,5±0,621,7±0,720,4±0,822,0±0,6

Before irradiation the body weight in the two groups was the same. After irradiation, the body weight of mice was decreased. Least this effect was pronounced in mice protected by the vaccine "Grippol" in a dose of 0.2 ml per 15 days of radiation sickness weight of their bodies was equal 22,4±0.4 g in control mice with 20.4±0,8 is. This difference is statistically significant. Student test t=2,2; p=0.04. The difference between the body weight of mice protected by the vaccine "Grippol" in a dose of 0.2 ml (22,4±0.4 g) or 0.05 ml (19,7±0.9 g)was statistically significant. Student test, t=2,6; p=0.01. Body weight of mice protected by the vaccine "Grippol" in a dose of 0.2 ml (22,4±0.4 g)was higher body weight of mice protected by the vaccine "Grippol" in a dose of 0.5 ml (21,5±0.7 g). However, this effect was less statistically significant than in the previous case. Student test, t=l,8; p=0,09. Thus, the optimal radioprotective dose of the vaccine "Grippol", judging by the dynamics of body weight, should be considered a dose of 0.2 ml.

When determining the optimal timing of administration of the vaccine "Grippol" to increase the radioresistance mice radioprotective dose of the vaccine of 0.2 ml was injected into mice for 7 or 14 days before irradiation. The results of the experiments are presented in table 2.

tr>
Table 2

The survival rate of mice protected by the vaccine "Grippol", administered at a dose of 0.2 ml for 7 or 14 days before irradiation by gamma-quanta137Cs at a dose of 7.5 G
A group of miceDates of vaccine administration before irradiation, dThe number of miceOf themSurvival,

(M±m,%)
Palosurvived
Experience14101990±10
Control-107330±15
Experience7123975±13
Control-105550±16

Table 2 shows that the maximum efficiency of the radioprotective action of the vaccine "Grippol" took place with the introduction of the vaccine at a dose of 0.2 ml for 14 days before irradiation at a dose of 7.5 Gy. In the protected group survived 90±10% of the animals in the control and 30±15%. The excess of the survival in the experimental group over the control was 60%. The effect is statistically significant, student test t=7,4; p=0.003. With the introduction of vaccines at least 7 days before irradiation in a protected group survived 75±13% of the animals in the control and 50±16%. The survival of protected mice is 25% higher survival rate in the control. However, this difference, according to the student test, statistically not significant.

Radioprotective effect of influenza vaccine "Grippol" was higher than that of previously investigated liquid inactivated vaccine Omutninsk chemical plant [3]. Radioprotective properties of the placenta is her were maximally expressed at the dose of 0.2 ml for 7 days before irradiation of 7.1 G (SD 24/30In this case survived 66,6±5.5% of mice in group-protected liquid inactivated vaccine, and 24.1±3.7% in the control group. The difference between the survival rate is 42.5 per cent. When using vaccine "Grippol" exceeding survival in the experimental group over the control was 60%. The results show that 14 days before irradiation, judging by the survival rate is the optimal time of administration of the vaccine "Grippol" for the manifestation of its radioprotective action.

Dose of 8.5 Gy resulted in the death of 100% of the control mice. Protection of mice with vaccine "Grippol" (0.2 ml of vaccine at least 14 days prior to the radiation damage) ensured the survival rate of 20% of the animals (table. 3).

Table 3

The average lifespan of mice protected by the vaccine "Grippol", administered at a dose of 0.2 ml for 14 days before irradiation by gamma-quanta137Cs dose of 8.5 Gy (SD100/30)
A group of miceThe number of miceOf themThe average lifespan, days (M±t)
Palosurvived
Experience108216,5±1,1
Control1010010,9�B1; 1,1

About the beneficial effects of the drug on the course of acute radiation sickness demonstrated by the increase in life expectancy protected mice. The average life expectancy of dead animals in the experimental group was equal to 16.5±1.1 days, and in control - 10,9±1.1 days. It follows that protection of mice with vaccine "Grippol" increased life expectancy of dead animals by 5.4 days. The difference between rates in the two groups is statistically significant. Student test, t=3,6; p<0,05.

Radioprotective effects of vaccines is manifested most clearly in the stimulation of the blood [8]. Radioprotective efficiency "Grippol" was evaluated by the effect on haematopoiesis in the bone marrow and peripheral blood cellularity in irradiated mice. Vaccine (0.2 ml) was administered subcutaneously for 14 days before irradiation by gamma-quanta137Cs at a dose of 6.0 Gy (SD10/30). Mice were scored on the 8th day after irradiation by decapitate under ether anesthesia. The study of the effects of the vaccine "Grippol" on the status of bone marrow hematopoiesis recorded the total number of cells in the bone marrow of the femur. On smears-prints of bone marrow stained Pappenheim [9], determined the content of the different generations of cells. The study of the effects of the vaccine "Grippol" on the state of the peripheral blood was determined containing the s of lymphocytes, neutrophils, platelets, reticulocytes, erythrocytes and hemoglobin level on the Hematology analyzer "Alson-6". The total number of leukocytes in blood were measured in the cell Goryaeva. Leukocyte counts were determined in smears stained Pappenheim. The results of the study status of hemopoiesis in mice are presented in tables 4 and 5.

0,6±0,1
Table 4

The impact of the vaccine "Grippol", administered at a dose of 0.2 ml for 14 days before exposure of mice at a dose of 6.0 Gy (SD10/30), the contents of the cells in the bone marrow of the femur (M±m×106)
The type of cells in the bone marrowThe group of animalsBefore irradiationAfter irradiation, the 8th day
Onlycontrol23,5±1,36,0±0,4
experience16,5±0,7*
erythroidcontrol5,2±0,41,0±0,4
experience7,1±0,6*
neutrophiliccontrol10,5±0,81,7±0,4
experience4,6±0,5*
lymphoidcontrol5,6±0,5
experience1,3±0,3*
Note: * -Statistically significant difference compared with control. Student test, p<0,05.

On the basis of the conducted researches it is established that in the process of development of acute radiation sickness in control mice, there is a sharp decrease in the content of cells in the bone marrow and peripheral blood. The introduction of the vaccine "Grippol" in a dose of 0.2 ml for 14 days before exposure had a significant effect on postradiation recovery of erythropoiesis, neytrofilopoeza and lymphopoiesis in the bone marrow, which was accompanied by increased levels of erythrocytes, reticulocytes, hemoglobin and neutrophils in peripheral blood. This is evidenced by statistically significant differences between experimental and control groups. It can be assumed that the basis of the radioprotective effect is the ability of the vaccine to provide a stimulating effect on post-radiation recovery of hemopoiesis.

Table 5

The impact of the vaccine "Grippol", administered at a dose of 0.2 ml for 14 days before exposure of mice at a dose of 6.0 Gy (SD10/30), the contents of the cells in the peripheral blood.
Blood cellsThe group of animalsTo irradiated what I After irradiation, the 8th day
neutrophils (×109/l)control0,9±0,10,1±0,02
experience0,3±0,04*
the reticulocytes (×109/l)control201,2±19,718,1±3,8
experience114,8±16,8*
erythrocytes (×1012/l)control9,9±0,38,3±0,1
experience9,3±0,2*
hemoglobin (g/l)control128,5±5,1105,4±2,2
experience119,1±1,8*
Note: * - Statistically significant difference compared with control. Student test, p<0,05.

Thus, our data strongly argue that the vaccine "Grippol" has a radioprotective effect - increases the body's resistance to radiation. It reduces the severity of acute radiation sickness and stimulate hematopoiesis. The data obtained indicate the possibility of using vaccine "Grippol" as drug double action:

anthropologi and radioprotective, for workers at the me industry and other groups at risk of radiation exposure.

Example 1

Radioprotective effects of the vaccine "Grippol" experience on mice hybrids (NEA×C57B1)F1. Are males weighing 18-23 g Vaccine "Grippol" injected mice as radioprotective funds once subcutaneously at a dose of 0.2 ml for 14 days before exposure. As a control use animals that are injected with saline. The randomization control and test groups were carried out by mass. In each group of 10 mice. Animals once are uniformly irradiated by gamma-quanta137Cs installation IGUR at a dose of 8.5 Gy (SD100/30). The dose rate of 1.2 rad/sec. Monitoring of dose rate and its spatial distribution is performed using thermoluminescent dosimeter X-AND using thermoluminescence aluminophosphate glass IP-7 error ±10%. Irradiation of mice is carried out in special plastic containers, divided into individual cells. The findings showed that the dose of 8.5 Gy results in the death of 100% of mice. The introduction of the vaccine "Grippol" in a volume of 0.2 ml for 14 days exposure ensured the survival rate of 20±12% of the animals. On the radioprotective effects of the vaccine "Grippol" demonstrated by the increase in life expectancy protected mice. The average life expectancy of dead animals in the experimental group equals 16.5±1.1 days, and in control - 10,9&x000B1; 1.1 day. Protection of mice with vaccine "Grippol" increased their life expectancy by 5.4 days compared with the control group. The difference between rates in the two groups is statistically significant, student test, t=3,6; p =0.003.

Example 2

The vaccine "Grippol" on haematopoiesis in the bone marrow and peripheral blood cellularity experience in mice hybrids (NEA×C57B1)F1. Are males, weighing 18-23 g Vaccine "Grippol" injected mice as radioprotective funds once subcutaneously at a dose of 0.2 ml for 14 days before exposure. As a control using mice, which are injected with saline. The randomization of the control and experimental groups is done according to body weight. In each group of 10 mice. Animals once are uniformly irradiated by gamma-quanta137Cs installation IGUR at a dose of 6.0 Gy (SD10/30). The dose rate of 1.2 rad/sec. Monitoring of dose rate and its spatial distribution is performed using thermoluminescent dosimeter X-AND using thermoluminescence aluminophosphate glass IP-7 error ±10%. Irradiation of mice is performed in special plastic containers, divided into individual cells. Mice score on the 8th day after irradiation by decapitate under ether anesthesia. When studying the state of kostovska the CSOs blood count the total number of cells in the bone marrow of the femur. On smears-prints of bone marrow stained Pappenheim, determine the content of myelokaryocytes and other cells of different generations. In the study of peripheral blood counts the number of cells (lymphocytes, neutrophils, platelets, reticulocytes, erythrocytes), determine the level of hemoglobin Hematology analyzer "Alson-6", and the total number of leukocytes in the camera Goryaeva. The leukocyte formula determined in smears stained Pappenheim. The observation results show that the dose of 6.0 Gy leads to a sharp decrease in the content of cells in the bone marrow of control mice. Before irradiation the total number of cells in the bone marrow of the femur was equal to 23.5±1,3 (×106); erythroid - 5,2±0,4 (×106); neutrophil - 10,5±0,8 (×106); lymphoid - 5,6±0,5 (×106). On the 8th day after irradiation in the control mice the total number of cells in the bone marrow of the femur is reduced to 6.0±0,4 (×106); erythroid - 1,0±0,4 (×106); neutrophil - to 1.7±0,4 (×106); lymphoid - 0.6±0,1 (×106). On the radioprotective effects of the vaccine "Grippol" indicates significantly less deterioration in the experimental group compared to the control animals. So, on the 8th day after irradiation in immunized mice, the total number of the notches in the bone marrow of the femur is equal to 16.5± 0,7 (×106); erythroid - 7,1±0,6 (×106); neutrophil - 4,6±0,5 (×106); lymphoid - 1.3=1:0,3 (×106). The difference between the compared groups are statistically significant, the criterion of Student, t=2,2-13,0; p<0,05-0,01.

The observation results show that the dose of 6.0 Gy leads to a sharp decrease in the content of cells in the peripheral blood of control mice. Before irradiation, the content of neutrophils is 0.9±0,1 (×109/l); reticulocytes - 201,2±19,7 (×109/l; erythrocytes - 9,9±0,3 (×1012/l); hemoglobin - 128,5±5,1(g/l). On the 8th day after irradiation in the control mice, the number of neutrophils is reduced to 0.1±0,02 (×109/l); reticulocytes - to 18.1±3,8 (×l09/l; erythrocytes up to 8.3±0; l(×l012/l); hemoglobin - up 105,4±2,2 (g/l). On the radioprotective effects of the vaccine "Grippol" shows a less pronounced decline in the experimental group compared to the control animals. So, on the 8th day after irradiation in immunized mice, the number of neutrophils was equal to 0.3±0,04 (×109/l); reticulocytes - 114,8±16,8 (×l09/l; erythrocytes - 9,3±0,2 (×1012/l); hemoglobin - 119,1±1,8 (g/l). The difference between the compared groups are statistically significant, the criterion of Student, t-4,4-5,6; p<0,01.

The sources of information is AI

1. Andrushchenko V.G., Ivanov A.A., V. Maltsev. Radioprotective action of substances of microbial origin (literature review) // Rad. biology. Radioecology. - 1996. - T.36. - B.2. - S-208.

2. Kamalan L.A. TO the question of the increase in radioresistance rabbits by immunization them vaccinia virus // Radiobiology. - 1966. - V.6. No. 6. - S-864.

3. Ivanov A.A., Ershov FI, Ulanova A. M., Kuzmina, ETC, Stavrakou NM, Tselakova AB, GA shalnova. Radioprotective and interferonogenna properties influenza vaccine // Rad. biology. Radioecology. - 1995. - T.35. - B.2.- S-235

4. RF patent №2164148 (IPC7A 61 K 39/145).

5. Instructions for use of influenza vaccine trivalent polymershapes liquid GRIPPOL. Registrazione ID No. 96.309.123. Approved by the Chief state sanitary doctor of the Russian Federation, Onishchenko 22.06. 2001

6. Khaitov R.M P.M., Nekrasov A.V., Gorbunov M.A. Vaccine "Grippol" the children's Vaccination. - 2001. - V.17. No. 5. - P.1-5.

7. Urbach VY Mathematical statistics for biologists and physicians // Publishing house of the USSR Academy of Sciences. - 1963. - 322 S.

8. Guzenko KK, Ivanov A.A., V. Maltsev. The effect of bacterial products on haematopoiesis irradiated animals // Rad. biology. Radioecology. - 1994. - T. - C.4-5. - S-586.

9. Coast E.A., the Handbook of clinical and laboratory research methods // M: - Medicine. - 1963. - 436 S.



 

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FIELD: organic chemistry, polymers, immunochemistry of high-molecular compounds.

SUBSTANCE: invention describes a synthetic hapten based on copolymers of N-vinylpyrrolidone, crotonic acid and p-crotonoyl aminophenol comprising covalently joined 2-naphthyl amine as a hapten of the following general formula:

wherein m = 88.5 mole%; n = 6.8-10.3 mole%, and l = 1.2-4.7 mole%. Invention provides specificity of antibodies raised against 2-naphthylamin as a hapten and possibility for preparing highly specific antibodies raised against carcinogen 2-naphthylamine as a hapten by immunization of rabbits with synthetic antigen comprising a copolymer of N-vinylpyrrolidone, crotonic acid and p-crotonoyl aminophenol as a macromolecular carrier. Also, invention provides possibility for practice using these antibodies for immunological recording 2-naphthylamine as a hapten in blood of industrial workers subjected for exposition with 2-naphthylamine. The proposed compound elicits antigenic features and can be used for detection of oncological risk among workers subjecting with contact with 2-naphthylamine.

EFFECT: valuable properties of antigen.

1 tbl, 1 dwg, 6 ex

FIELD: medicine.

SUBSTANCE: vaccine is high molecular weight protein conjugate with angiotensine II taken in high molecular weight protein : angiotensine II proportion of 1:12-55 in % by weight. The conjugate is modified with equilibrium quantity of immunocompetent polyelectrolyte like polyoxydonium.

EFFECT: stable physiological response within prolonged period of 6-12 months.

3 tbl

FIELD: immunology.

SUBSTANCE: the innovation deals with new immunogenic conjugates of beta-propionamide-bound polysaccharide and N-propionamide-bound oligosaccharide with protein, and the method to obtain these conjugates has been suggested, as well. Conjugates should be applied to obtain vaccines against infectious diseases and cancer that enables to broaden the number of preparations applied in treating the above-mentioned diseases.

EFFECT: higher efficiency.

1 dwg, 2 ex, 8 tbl

The invention relates to medicine, in particular to immunology, and can be used for the treatment of drug addiction, poisoning with various poisons in industry and everyday life, when man-made disasters, etc

The invention relates to medicine and relates to methods of treatment of tumors and metastases using a combination of antiangiogenic therapy and immunotherapy

The invention relates to the field of medicine and relates to a composition of recombinant proteins, the method of obtaining the composition, the pharmaceutical kit of reagents for immune therapy and preventive vaccination of tumor diseases anogenital areas and method of immunotherapy and preventive vaccination

FIELD: medicine, in particular immunogenic composition.

SUBSTANCE: claimed composition contains antigens of A and B influenza viruses, sorbed on Bifidobacterium bifidum bacterial cultures, immobilized on acceptable sorbent.

EFFECT: increased immunogenicity.

2 cl, 2 ex, 5 tbl, 2 dwg

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