RU2396978C1 - Method for prevention of cattle leukosis development - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology. The method involves vaccination of animals with the vaccine prepared by inactivation of cultural leukosis virus and polymerisation of its glycoproteides with 0.2% formalin at 39-40°C for 7-9 days and aluminium hydroxide sorption of the prepared suspension in amount 3-5 mg/ml. The vaccine is introduced subcutaneously, three times, every 14-15 days, in amount 3-5 ml. Herewith, 20-30 days prior to vaccination, tuberculosis anatoxin 4-5 ml and staphylococcal anatoxin-vaccine 4-5 ml are introduced subcutaneously. 8-9 months later, revaccination is performed as a single introduction of the leukosis vaccines preceded with introduction of tuberculosis anatoxin and staphylococcal anatoxin-vaccine in specified amount 20 days before.

EFFECT: method provides more effective protection of the animals vaccinated against leukosis.

2 cl, 3 ex

Description

The invention relates to virology, immunology and biotechnology, in particular to methods for the prevention of cattle leukemia.

Known methods for producing inactivated vaccines, toxoids, allergoids (tolerogens) include detoxification and polymerization of toxins, inactivation of viruses, bacteria with formalin, glutaraldehyde, hydrogen peroxide, etc. (Burchasov P.N. Guidelines for vaccine and serum production. M: Medicine, 1978. - P.72-87; Fradkin V.A. Diagnostic and therapeutic allergens. M: Medicine. 1990. - 256 s; Vorobev A .A. Microbiology and Immunology. M.: Medicine. 1999. - S.260-266).

Existing methods for producing an inactivated vaccine for the prevention of cattle leukemia include the cultivation of leukemia virus in cell culture and inactivation with formalin and others with the subsequent addition of adjuvant or the production of live vaccines based on the vaccinia virus (Syurin N.V., Samuilenko A.Ya. and Viral diseases of animals. M.: VNITIBL, 1998. - P. 400-406; Busol V.A., Shapovalova O.V., Nagaeva L.I. et al. Vibration of the overwhelming effect of the reaction. I have inactivated the vaccine against leukemia veliko horny thinness on vivc x i telits // Veterinary Medicine. // Mizhvidchy themed science zbirnik. - K .: 1995. - Issue 70. - P.3-9; Zaviryukha A.I., Zaviryuha G.A. et al. Results of vaccination vaccine “Leukemia” in prevention and combating leukemia with great horny thinness // Veterinary Medicine. Mid-sized thematic science zbirnik. - X., 2003. - Vol. 82. - P.240-246; Krikun VA, Gulyukin M.I. The problems of improving farms unsuccessful in leukemia and the prospects for developing specific prophylaxis of cattle leukemia // Tr. VNIIEV. - T.72. - M., 1999 .-- P.49-52).

However, the use of vaccines for the prevention of cattle leukemia to a certain extent has not received wide practical use, mainly due to the lack of means for the correction of immunological deficiency and the formation of antibodies with a low titer.

Therefore, improving the production of whole-virion and subvirion vaccines for the prevention of leukemia should be interconnected with the simultaneous use of specific immunomodulators. The solution to this problem was and remains relevant for both veterinary and humane medicine.

The prototype is “The way to get the vaccine against leukemia of great horny skin thin” (Busol V.A., Shapovalava O.V., Myagky N.V. et al. Patent on wines No. 35439, A61K 39/21, dated October 14, 1999 )

The disadvantage of this method of obtaining a vaccine against leukemia in cattle, as well as other similar vaccines: "Leukopol", "Leukozav" and others, is the lack of manufacturing technology for the vaccine, the process and means of polymerization of fragments of different virus structures, a control infectious dose for vaccinated animals, the period of revaccination and data of the immunological state of animals before vaccination, taking into account depressive environmental factors, the need to stimulate humoral or cellular immunity.

The effectiveness of a number of drugs that affect the immune system in cattle with leukemia, such as levomizole, unitiol, roncoleukin, lycopid, mixoferon, ASD, chitosan, catozal, etc., has not been studied and they have no practical application.

In our studies, the developed and tested toxoid from mycobacterium tuberculosis for the prevention of tuberculosis, the toxoid vaccine for the prevention and treatment of staphylococcosis subsequently showed effectiveness in oncogenic pathologies in humans and carnivores, both in a single application and in combination.

Based on the fact that the staphylococcal toxoid vaccine and tuberculous toxoid provide a normal dosage of the humoral and cellular parameters of the immune system at a certain dosage, studies were carried out in combination with the inactivated leukemia vaccine for the prevention of cattle leukemia.

To eliminate these disadvantages of the prototype, vaccination of animals is proposed by subcutaneous injection of 3-5 ml of the leukemia vaccine three times with an interval of 14-15 days with a preliminary, 20-30 days, subcutaneous injection of tuberculous toxoid and staphylococcal toxoid vaccine 4-5 ml as immunostimulants , and after 8-9 months, a single booster vaccine against leukemia with a preliminary, for 20 days, the introduction of the above immunostimulants in the previous volumes.

The preliminary combined use of tuberculosis toxoid and staphylococcal toxoid vaccine increases the effectiveness of the inactivated whole-virion vaccine against cattle leukemia and protects vaccinated animals from infection and prevents the development of pathologies characteristic of leukemia when vaccinated and leukemic cows are kept together. Moreover, the titer of precipitating antibodies for 15-16 months was at the level of 1:16.

The aim of the invention is to increase the effectiveness of prevention of the development of cattle leukemia by vaccinating it with an inactivated anti-leukemia vaccine and increasing its protective properties by simultaneously administering the staphylococcal toxoid vaccine and tuberculous toxoid as immunostimulants.

This goal is achieved by the introduction of a vaccine obtained by inactivation of the culture virus of leukemia and polymerization of structural fragments of the virus (glycoproteins) with 0.2% formalin solution at 39-40 ° C for 7-9 days, sorption on aluminum hydroxide (3-5 mg of aluminum hydroxide per 1 ml of suspension). The resulting vaccine is administered 3 times subcutaneously to heifers, cows in a volume of 3-5 ml with an interval of 14-15 days with a preliminary, 20-30 days before vaccination, subcutaneous injection of tuberculous toxoid and staphylococcal toxoid vaccine in a volume of 4-5 ml of each drug . After 8-9 months, revaccination is carried out using a single injection of immunomodulators and after 20 days of the vaccine in the above volumes. At the same time, purulent abscesses do not form at the injection site of immunomodulators and the vaccine, and the antibody titer in the RID is manifested in a 1:16 dilution.

It is also worth noting another positive effect of these drugs before or after removal of the prostate, breast, with bladder cancer and fibroma with subcutaneous administration with an interval of 5-7 days for 1.5-2.0 months.

The high protective efficacy in the prevention of cattle leukemia and the positive results of immunotherapy of certain forms of cancer in humans with a leukemia vaccine with a staphylococcal toxoid vaccine and tuberculous toxoid instead of BCG create the prospect of their use in veterinary and humane medicine.

In the patent and scientific and technical literature, no technical solutions were found that were similar to those declared using staphylococcal toxoid vaccine and tuberculous toxoid.

The use of an inactivated culture vaccine for the prevention of cattle leukemia with a staphylococcal toxoid vaccine and tuberculous toxoid allows to achieve the protective effect of the vaccine specified in the purpose of the invention.

The established possibility of inactivation of the culture virus of leukemia and the polymerization of structural fragments of the virus (glycoproteins) with 0.2% formalin solution at 39-40 ° C for 7-9 days, sorption on aluminum hydroxide (3-5 mg / ml) can be used for production cattle leukemia vaccines in an industrial setting.

The results of the preparation and use of an inactivated vaccine against leukemia in cattle with the preliminary administration of the staphylococcal toxoid vaccine and tuberculous toxoid previously developed and tested by us are presented in the following examples.

Example 1. A method of obtaining a vaccine against leukemia

The cattle leukemia vaccine was obtained by inactivating 1 liter of a suspension of culture leukemia virus and polymerizing the glycoproteins of the virus with 0.2% formalin solution at 39-40 ° C for 9 days, sorption on aluminum hydroxide (3-5 mg / ml), followed by packaging in sterile vials and sterilization at 75-80 ° C for 30-40 minutes.

Example 2. The vaccine test for harmlessness

The vaccine was tested harmlessly by administering the vaccine 3 times subcutaneously to 12 calves and 14 heifers of 4-6 months of age in a volume of 5 ml with an interval of 10 days.

In vaccinated animals, there was no increase in temperature and anxiety in the absence of the formation of purulent abscesses at the injection site.

Example 3. The test vaccine for protective properties

In studies, 25 heifers and 18 cows of a dysfunctional leukemia farm were used, in which the antibody titer in RID was 1: 2 and 1: 4, but there were no pronounced pathological changes in the organs and clinical signs characteristic of leukemia.

Before vaccination, all animals were injected subcutaneously with 5 ml of staphylococcal toxoid vaccine and tuberculous toxoid, and then after 20 days the animals were vaccinated with a leukemia vaccine in a volume of 4-5 ml by subcutaneous injection three times in the middle third of the neck with an interval of 14-15 days from subsequent revaccination after 9 months, in which the leukemia vaccine was injected subcutaneously with preliminary, 20 days before vaccination, subcutaneous administration of tuberculous toxoid and staphylococcal toxoid vaccine in the previous their volumes.

Over the course of 15 months, the vaccinated animals did not reveal hematological changes and pathological lesions in the lymph nodes, spleen, and clinical signs characteristic of leukemia. At the injection site, the formation of purulent abscesses was not noted. Moreover, in vaccinated animals, the titer of precipitating antibodies in the RID was at a constant high level of 1: 8-1: 16, and in patients with cows leukemia this titer was 1: 2-1: 4.

Claims (2)

1. A method for preventing the development of cattle leukemia, including the introduction of an inactivated vaccine against leukemia, characterized in that the animals are vaccinated with a vaccine obtained by inactivating the culture of leukemia virus and polymerizing its glycoproteins with a 0.2% formalin solution at 39-40 ° C in for 7-9 days and sorption of the resulting suspension on aluminum hydroxide 3-5 mg / ml, the vaccine is administered subcutaneously, three times, with an interval of 14-15 days, in a volume of 3-5 ml, while previously subcutaneously 20-30 days before vaccination introduce that erkulezny staphylococcal toxoid and toxoid vaccine on 4-5 ml and 8-9 months after revaccination performed by single administration of a vaccine with leukemic preliminary 20 day administration tuberculous staphylococcal toxoid and toxoid vaccine in the previous levels. 2. The method according to claim 1, characterized in that the vaccine is subjected to infected animals with a titer of precipitating antibodies of 1: 2-1: 4.