" RMZH, Volume 9 № 16-17, 2001
Study of the immunomodulating activity of Arbidol
K.m.n. YE.P. Of sel'kova, I.YU. Gracheva,
k.m.n. T. P g.otvyan.chk.aya, Danilin's k.m.n. G.A.,
k.m.n. M.P. Amaryan, d.m.n. T.A. Semenenko
GKB im. S.P. of Botkin, GKB № 3, g. Zelenograd
At present there does not exist vaccine preparations against the urgent agents of sharp respiratory virus infections (ORVI) (with exception of influenza vaccines), which contributes to the maintenance of morbidity at the high level. In connection with this to completely regularly accentuate attention to the approval and application of new effective means of unspecific preventive maintenance ORVI. One of the methods of solution of the problem indicated is the search for the preparations, which combine antiviral activity with the immunostimulating properties. Unspecific preventive maintenance is shown to people with the clinical signs of the decreased resistance of the organism: _ frequent respiratory disease with protracted or complicate flow; by the presence of the chronic centers of infections; by severe chronic somatical illnesses [ 1 ]. Not is less urgent unspecific preventive maintenance ORVI among the medical workers, in view of the specific character of its activity who are the group of professional risk.
The purpose of the present investigation was the study of the immunomodulating activity of the domestic preparation of arbidol, created in the Center for chemistry of medicines - All-Russian scientific research chemicopharmaceutical institute (VNIKHFI - ALL-UNION SCIENTIFIC RESEARCH CHEMICAL AND PHARMACEUTICAL INSTITUTE IM. SERGO ORDZHONIKIDZE); and permitted Farmkomitetom MZ RF to the clinical application in adult [ 2 ].
Materials and the methods
The study of the influence of arbidola on immune status was conducted on the limited contingent of medical workers TSKB №y g. of zelenograda. By the method of the random sampling were formed 2 equivalent in every respect groups of 38 people in each: the 1st - obtained arbidol (experimental); 2-4 - placebo according to the same diagram (control group). Arbidol in the form of tablets on 0.2 g used orally 2 times a week during 3 weeks. Research were conducted in the dynamics: prior to the beginning of the application of preparation (I test), in 7 days (II test) and in 21 days (III test) from the beginning of the application of arbidola.
The immunomodulating activity of preparation was determined in accordance with the systematic recommendations regarding the estimation of immune status of man, the developed colleagues of the institute of immunology MZ RF under the management by Acad.. WOUNDS R.V. Of Petrov [ 3 ]. All persons in the experimental and control groups were inspected according to the united diagram, which included the determination of the number of leukocytes and lymphocytes, T-lymphocytes (CD3) and their subpopulations (CD4 and CD8), immunoregulator index (CD4/CD8), content of B-lymphocytes (CD19), NK- cells (natural killers - S.DY'), immunoglobulins of basic classes, TSIK and indices of phagocytosis, i.e., the parameters, which reflect the functioning of basic it was sectional immune system (cellular, humoral and phagocytic).
For the evaluation of the influence of preparation on the indices of specific immunity a serological study of the models of blood serum is carry ouied. By the randomized method were formed 2 groups: experimental (96 people) obtained arbidol, control room (107 people) - placebo. Blood serum were investigated to the presence of specific antibodies to the viruses of influenza A (H1N1 and H3N2) and in, to respiratory-syncytial (PC) virus, to adenovirus and to the viruses of parainfluenza of the 1st and 3rd types in the immunoenzyme analysis (IFA). With setting of reaction was used the test- system, which is the 13- component collection of the antigens of the respiratory viruses, utilized for the absorption on the surface of the holes of polystyrene plane-tables, and also the conjugates of monoclonal antibodies to IgM and IgG man (production OF NII - SCIENTIFIC RESEARCH INSTITUTE influenza IT WAS FRAME). The detection of specific complex antigen-antibody was produced with the aid of the marked by peroxidase monoclonal antibodies to IgM and IgG man.
Results and the consideration
The conducted investigations showed that arbidol renders the immunomodulating action, leading to an increase in the total quantity of T-lymphocytes and T- helpers, which was determined with the aid of the monoclonal antibodies to the antigens of the lymphocytes (see table).
It should be noted that in this case the discussion deals with the normalization of indices in the persons with that initially decreased clean CD3 and CD4 cells, in the absence of influence on the total quantity of T-lymphocytes and T- helpers in the persons with the normal functioning of the cellular component of immunity. Egi given will agree with the results, obtained earlier in the limited experiences by other authors [ 4 ].
At the same time the application of arbidola did not lead to substantial changes of the absolute number of T- suppressor cells as a whole on the groups, which confirms the opinion of the number of researchers about the fact that arbidola stimulating activity is not connected with the oppression of the function of suppressor cells [ 5 ].
Determination of immunoregulator index (relationship of T- helpers and T -supressorov) also gives important information about the state of the immunity of man, since his decrease can testify about development or presence of immunodeficient state. Obtained by us given testify about the absence of the action of arbidola on the immunoregulator index, which is located within the limits of standard, and its normalizations in 11 people with the initially decreased indices.
The important property of arbidola is its stimulating influence on the phagocytosis. In the experiments carried out earlier on animals it was shown that the preparation not only increases the total number of macrophages with the absorbed bacteria, but also increases the phagocytic number [ 6 ]. Strengthening phagocytosis is noted in patients, who suffer chronic bronchitis, which was accompanied by an improvement in the clinical state of the sick and in a number cases it made it possible to forego the method of antibiotics [ 7,8 ]. In the conducted by us investigations confirmed stimulating action arbidola on the phagocytic component of immunity; the ensuring increase in the stability of macroorganism. The determination of the phagocytic activity of neutrophils with the aid of the microscopic calculation of the phagocytizing cells and phagocytized units made it possible to establish a statistically reliable increase in the phagocytic index in those, who obtained arbidol (from 49,2% to 67,8% and 76,1% when II and III inspection, correspondingly, of r<0,0shch).
These data have important significance, since precisely phagocytosis is the condition of the realization of intracellular cytolysis, most effective with the protection from the infectious agents. In all likelihood, the activating stimuli for the phagocytic cells in the persons of experimental group were tsitokiny and, in particular, interferon, whose production is strengthened under the action of arbidola [ 6 ].
In the persons of experimental group the increase in the content of natural killers, especially expressed with the second inspection, was noted. In contrast to the control room in the experimental group with the specific fluctuations of the initial levels NK- cells the difference in the indices to (11±0,9%) and after the method of arbidola with II inspection (16±1,1%) and III inspection (13±1,2%) was significant and statistically reliable (p<0,05), which makes it possible to characterize preparation as the inductor of the activity of natural killers.
In the process of experiments it was impossible to establish the essential influence of arbidola on the content of B-lymphocytes and the levels of the immunoglobulins of basic classes. Absence in arbidola of the immunosuppressing effect on the production of specific antibodies to the respiratory viruses was confirmed the results OF IFA before the method of preparations and 3 months later, i.e., after the end of the period of the seasonal lift of morbidity. The application of arbidola did not exert a substantial influence on level and dynamics of the formation of the specific antibodies: the statistically reliable differences between the indices of experimental and control room of groups it is not revealed. The uniform appearance of antibodies in the blood serum of the investigated persons depending on circulation in the association of different viruses is established. In connection with this it is not possible to make a conclusion about the suppressing influence of preparation on the production of specific antibodies with the infection by agents ORVI. These data will agree with the opinion also of a number of other authors [ 6,8 ].
The simultaneous determination of the diagnostic captions of antibodies to two or more infectious agents, which were being evaluated at a study, it is noted in 43,7±5,1% (experimental group) and 52,2±5,4% of cases (control group). The specific weight of the separate exciters, which enter in the association, was unequal and depended on the intensity of their circulation in the period of inspection. The most frequent "partners" with the formation of the mixed infections both in the experimental and control group were the viruses of 1 types parainfluenza (27,8±5,5% and 25,6±4,1%) and adenoviruses (18,8±3,6% and 18,4±2,3%). Frequently they were encountered in the association the virus of influenza A (H3N2) (9,5±2,6% and 10,3±1,7%) and the virus of 3 types parainfluenza (6,9±2,6% and 7,8±1,2%), respectively.
Thus, the results of the conducted investigations attest to the fact that arbidol possesses the immunomodulating activity, which is manifested in an increase in the total quantity of T-lymphocytes and T- helpers, stimulation of the phagocytic function of neutrophils and induction of the activity of natural killers. Preparation does not render the immunosuppressing effect on the production of specific antibodies to the respiratory viruses, which profitably distinguishes it from the majority of the antiviral means, which are adapted for preventive maintenance and treating of influenza and ORVI.
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