ДЕЙСТВИЕ СИНТЕТИЧЕСКОГО АДЪЮВАНТА НА ФОРМИРОВАНИЕ ИММУННОЙ РЕАКЦИИ

Научная статья
DOI:
https://doi.org/10.18454/IRJ.2016.50.013
Выпуск: № 8 (50), 2016
Опубликована:
2016/08/18
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Савина С.В.1, Иващенко С.В.2, Скорляков В.М.3, Муртаева В.С.4

1 Кандидат ветеринарных наук, 2 Кандидат биологических наук, 3 Доктор ветеринарных наук, ФГБОУ ВО Саратовский аграрный университет им. Н.И. Вавилова

ДЕЙСТВИЕ СИНТЕТИЧЕСКОГО АДЪЮВАНТА НА ФОРМИРОВАНИЕ ИММУННОЙ РЕАКЦИИ

Аннотация

Статья содержит экспериментальные материалы по действию синтетического адъюванта на формирование иммунных реакций.

В настоящее время продолжается поиск новых эффективных вакцинных препаратов на основе полимерных соединений, обладающих иммуностимулирующими свойствами (Петров Р.В. и др.1986 г. Schijns V.,2000 г.)

Важным условием в разработке определённых носителей (адъювантов) является создание препарата способного долговременно находится в организме, с его активным состоянием иммунной системы, без местных и общих отрицательных реакций в организме (Вольпина О.М.,1992 г. и др.).

Нами проведены эксперименты по изучению синтетического адъюванта (АД), исходным сырьем которого является полидиаллилдиметиламмоний хлористый - ПЭС.

Проведенные доклинические испытания по изучению острой и хронической токсичности на лабораторных животных показали отсутствие отклонений в физиологических и  токсических параметрах, т.е. его безвредность.

Результаты показывают положительное влияние синтетического адъюванта на формирование общих и иммунных реакций в организме животных, что позволяет его использовать как наполнитель в вакцинах.

Ключевые слова: адъювант, вакцина, иммунная система, антитела.

Savina S.V.1, Ivashchenko S.V.2, Skornyakov V.M.3Murtaevа V.S.4

1 PhD in Veterinary Medicine and Science, 2 PhD in Biology, 3 PhD in Veterinary Medicine and Science, Saratov agrarian University n.a. N.I. Vavilov

THE EFFECT OF SYNTHETIC ADJUVANT ON THE FORMATION OF THE IMMUNE RESPONSE

Abstract

The paper contains experimental materials on the effects of synthetic adjuvants on the formation of the immune responses.

Currently, the search continues for new effective vaccines based on polymeric compounds having immunostimulating properties (Petrov R. V. et al. 1986, V. Schijns,2000)

An important condition in the development of certain carriers (adjuvants) is the creation of a drug capable of long-term is in the body, with its active immune system and no local and General negative reactions in the body (Volpin O. M.,1992, etc.).

We carried out experiments to study the synthetic adjuvant (BP), the raw material which is polydiallyldimethyl chloride - PES.

Conducted preclinical trials to study acute and chronic toxicity in laboratory animals showed no abnormalities in physiological and Toxicological parameters, i.e. its harmlessness.

The results show positive effects of synthetic adjuvants on the formation of General immune responses in animals that allow it to be used as a filler in vaccines.

Keywords: adjuvant, vaccine, immune system.

Vaccination in modern conditions is considered as one of the most effective means of infection control (Vogel F. R., 2000).

Adjuvants (BP) are used in vaccines to enhance the immune response for more than 80 years. Development of adjuvants and adjuvant systems has passed a way from the first empirical experiments to create purposeful systems, which is caused mainly by the progress in the study of the immune system and improvement of analytical, chemical and immunological methods (Vogel F. R. ,1998).

When creating artificial vaccines need to be artificial Bioorganic complex, which would ensure the body's immune response to the antigen against his genetically overcome low otvechaete. Only in this case, an artificial vaccine will be a significant step forward compared to the natural, which is a preparations of weakened or killed infectious agents or drugs selected them antigenic substance. ( Petrov R. V., Khaitov R. M.,1979).

Currently, as HELL in vaccines most commonly used aluminium compounds. They demonstrated its efficacy and safety for millions of administered doses of vaccines. In addition to these, as HELL in the production of vaccines uses other substances such as emulsions, liposomes and virosome. However, not rarely marked local reaction (8-10 %) for the introduction of such a HELL.

Creation of new effective vaccines, is associated with the search for polymeric compounds having immunostimulating properties (Petrov R. V., Kabanov V. A., Khaitov R. M., 1986)

M. SELA, in 1974, was the first to describe the artificially produced peptide causing the formation of antibodies to the protein lysozyme.

An important condition in the development of certain carriers (adjuvants) is the creation of long-term drug is able to be in the body, with its active immune system and no local and General negative reactions in the body (Vorobiev A. A., Vasiliev A. A., 1969; Volpin O. M., 1992)

In subsequent decades, the research Institute laboratories has been a different approach to the problem of synthetic vaccines. Its essence lies in the fact that artificial vaccines need to carry on their macromolecules, not only the determinant fragments of antigens, but also a structure that would ensure the production of antibodies to diverse antigens.

 In the study of the mechanism of action of synthetic polyelectrolytes (PES) on immunogenesis it was found that carbon-chain polyelectrolytes poly-4-vinylpyridine and poly-acrylic acid (PVA, HSC) stimulate the proliferation and migration of hematopoietic stem cells, the resettlement of T-lymphocytes from the thymus and b-lymphocytes from the bone marrow into the peripheral lymphatic organs and causes a significant increase in output in emerging antiteloobrazovania (Petrov R. V., Khaitov R. M. R. V. 1979).

Thus, the introduction into the organism of the polyelectrolyte and antigen during its active interaction with component of the blood and lymphoid system contribute to the establishment of long intercellular, intermolecular contact, which leads to increased antigenstimulated answer.

When you conduct a comparative evaluation of different types of adjuvants : aluminum hydroxide (D), ISCOM and Montanide Gel on the antigenic activity of rabies vaccine from strain ERA-CB 20 M as in the experimental rabies of single or and as part of polyvaccine in small animals it is established that the maximum titer of rabies antibodies (14,58 IU/ml ) was observed after application of the ISCOM adjuvant in the rabies monovaccine, with the use of the ISCOM vaccine in multivalent antibody titre was 7.2 IU\ ml (Kahnovich M. A., etc.)

The object of our research is a synthetic adjuvant (BP), the raw material which is polydiallyldimethyl chloride - PES.

Conducted preclinical trials to study acute and chronic toxicity HELL on laboratory animals showed no abnormalities in physiological and Toxicological parameters, i.e. safety.

Intramuscular adjuvant for 5 days did not cause any changes in the condition of the animals, hematological indices and internal organs (autopsy).

Conducted research locally irritating and allergenic (sensitising) steps in the formulation of conjunctival and skin tests. When visual assessment of the conjunctiva, cornea and eyelids found that AD does not cause irritation of mucous membranes of eyes and do not possess allergenic (sensitising) effect.

Conducted skin tests did not reveal damage in the form of erythema or edema. The application of the preparation did not cause redness, desquamation, allergic eczema and dermatitis.

The results showed that the polyelectrolyte is not mutagenic according to AMES test results, not teratogenic (does not cause malformations and deformities), NOEL= 25 mg/kg.

Tests performed at the research Institute for veterinary Virology and Microbiology (Moscow region, Cover) on inaktivirovannoj the antigen of rabies according to the one-HUNDRED 00495549-0020-2006 the standard scheme. According to the technical regulations revealed a high immunogenic activity in groups of 3 (Test vaccine + AD) 4-th (Reference vaccine + BP) and 5th (Inactivated antigen without GOA + AD) respectively of 1.82 IU/cm3; of 2.18 IU/cm3; of 2.26 IU/cm3

The experiments on the evaluation of the activity of the production of antibodies in the serum of rabbits immunized with disintegrated membranes (DM) Y. pseudotuberculosis by using the HELL. DM received successive destruction of the bacterial mass of Yersinia pseudotuberculosis microbe with an ultrasonic disintegrator, centrifugation, treatment with a solution of sodium dodecyl sulphate and dialysis. The preparation was concentrated with a rotary evaporator, the protein content 2mg in 1ml.

Rabbits had immunizable subcutaneously along the back of 3-4 points in a volume of 1 ml of a mixture of DM and BP in a 1:1 ratio. The antigenic dose of the drug was 1 mg protein per rabbit. Held 3 immunization at intervals of 2 weeks. Blood for research was taken from the auricular vein in a volume of 5-10ml per day before the introduction of the antigen. As control was used a group of rabbits, which were injected: DM + complete Freund adjuvant (AF), physiological solution (FR) + HELL, the DF + AF.

The antibody activity of the obtained sera was studied in indirect solid-phase enzyme immunoassay on plates (ELISA) with a DM of Yersinia pseudotuberculosis microbe in a concentration of 1мкг/ml (table.1).

The antibody titer of the blood serum of rabbits immunized with DM + AD made: after the 1st immunization 1:400 after 2nd - 1:51200, after the 3rd - 1:51200 (table.1). Thus, there was a sharp increase in the number of antibodies to the 2nd immunization in 128 times.

Table 1 - Results of determination of antibody activity of the sera obtained in ELISA with disintegrated membranes of Yersinia pseudotuberculosis microbe (1мкг/ml) after the third immunization

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Thus, the preclinical testing acute, chronic toxicity, hematological, biochemical processes and determination of antibody activity have shown positive effects of synthetic adjuvants on the formation of General immune responses in animals that allow it to be used as a filler in vaccines.

References

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