Abstract

Purpose: In recent decades, the activity and points of influence of probiotics, microelement compounds and vitamins, which play a significant role as cofactors in maintaining the immune system, both local and systemic, specific and innate, at the level of stable homeostasis, have been intensively studied. Given the insufficient scientific information on research in this direction, the aim of the research was to determine the effect of prebiotics on the production of anamnestic antibodies to viruses and microbes of the respiratory group by cells of the palatine tonsils of patients with chronic tonsillitis in an in vitro experiment.

Material and methods: The study was conducted on tonsil cell cultures obtained from 20 patients with chronic tonsillitis who underwent tonsillectomy according to indications. The age of the examinees ranged from 16 to 40 years, of which 14 were female. Cells in the control and experiment were cultivated at 37⁰C for 24 hours. Various prebiotic preparations were added to the experimental samples before cultivation. To the culture of tonsil cells to each one separately was added Vit C in a dose of 50 μg, D₃-aquadetrim – 25 units, zinc solution – 50 μg. 2 μg of normal human serum protein was added to the control samples. After 24-hour cultivation, supernatants were ob tained, in which antibodies to respiratory viruses were determined: to influenzaviruses A- and B- by the reaction of inhibition of hemagglutination of erythrocytes of the AB group, to N- and S – antigens of the SARS-CoV-2 virus, to the capsid antigen of the EBV virus (Euriimmen), to Human orthopneumovirus, to herpes simplex virus type 1 (HSV-1) by ELISA method, to streptolysin-O of Streptococcus pyogenes (ASL-O) by latex method and proteglycans Klebsiella pneumoniae by reaction of indirect hemagglutination according to Boyden. The statistical processing of the obtained results was carried out using the one-sided non-parametric “U” test of Mann-Whitney, and the parametric t-Student method was used in the analysis of log₂ titers of AB. Calculations were made using WIN PEPI and Biostat open access software packages

Results: It was established that no significant reactions of tonsil cells were detected to all the drugs used in the content of antibodies to antigens of the SARS-CoV-2 and VEB viruses. Hemagglutinins of the influenza virus were detected in significant titers (1:16 or more) in 17% of studies, under the influence of zinc and vitamin D₃ preparations – in 26% of supenatants. ASL-O was actively produced under the influence of vitamin C in 33%, D₃ – in 25%, and zinc in 50% of all examined tonsilocyte cultures. In addition, the content of antibodies to Human orthopneumovirus and HSV-1-antigens in the culture fluid increased under the influence of the trace element zinc. According to the sum of positive effects, this trace element was the most active among the studied prebiotics. The low level of stimulation of the content of antibodies to K. pneumonia by these prebiotics may partially indicate a low level of infection and the low-active formation of immunological memory cells in the palatine tonsils.

Discussion: It can be assumed that the palatine tonsils have a rather variable potential in terms of an antibody-type reaction with the presence of antigens competition, and in this case, the studied antibody-producing cells prevail in the anamnestic reaction to antigens of influenza and S. pyogenes , the anamnestic humoral reaction to VEB and N- and S- antigens of the SARS-CoV-2 virus was much less pronouncedThe obtained data indicate the possibility of stimulation of cells of the palatine tonsils of patients with chronic tonsillitis with regard to antibody production against antigens of respiratory viruses with the use of vitamins D₃, C, trace element zinc.

In terms of the perspective of the work, there are two directions of research. The first is the determination of the effect on antibodyogenesis against viruses and microbes of an additional amount of prebiotics and mixed complexes of various prebiotic preparations. The second is the determination of the effectiveness of prebiotics during intranasal vaccination using nanoparticles, which is a new approach in changing the existing paradigm of parenteral vaccination. Further conducting of such studies will contribute to the creation of preclinical models of vaccines against viruses of the respiratory group of a new type for local (intranasal and oral) use in the near future.

Conclusions: In vitro experimental studies on determining the effect of viral antigens and prebiotics on the immunological potential of cells of the palatine tonsils are a reliable and quick method of approximate assessment of the degree of immunotropy of drugs of different groups. Cells of the palatine tonsils in vitro responded by increasing the level of antibodies to the antigens of influenza A and B viruses, Human orthopneumovirus and HSV-1, to streptolysin-O of S. pyogenes under the influence of the drugs introduced into the cultures. The use of the trace element zinc appears to be the most active. Positive changes regarding the level increase of antibodies to virus antigens were also noted under the action of vitamins D₃ and C.

Key words: palatine tonsils, immunological potential, prebiotics, respiratory infections.