Evaluation of immune response of BALB/c to homeopathic solutions
Keywords:Biotherapic, Influenza, flow cytometry, immunehistochemistry, animal model.
AbstractIntroduction: Biotherapics are medicines prepared from etiologic agents, following Brazilian Homeopathic Pharmacopeia. Influenza is a disease that affects thousands of people worldwide every year, motivating the development of new therapies. Aim: In this study, we developed two biotherapics from live/active influenza A virus, at 12x and 30x, and verified some immune response parameters in mice. Methodology: The biotherapic was administered to male SPF 4 weeks old Balb/c mice. The protocol was approved by the UFRJ Ethics Committee of Animal Use (Protocol DFBCICB 040). Animals were stimulated daily, blindly, with different homeopathic medicines, at 1% (V/V) for a maximum period of 42 days. Three homeopathic medicines were tested: biotherapic 30x containing active influenza A virus; biotherapic 12x containing active influenza A virus; and thymulin 5cH. The experimental groups were: Group A (5 animals) ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ administration of thymulin 5cH, Group B (5 animals) ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ administration of biotherapic 30x, Group C (5 animals) ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ administration of biotherapic 12x, Group D (5 animals) - administration of a water 30x, Group E (5 animals) - administration of a water 12x, Group F (5 animals) - control (without treatment). After 21 days of treatment, all animals were challenged subcutaneously with the viral hemagglutinin antigen at the concentration of 7 ÃƒÂ¯Ã‚ÂÃ‚Âg/200ÃƒÂ¯Ã‚ÂÃ‚ÂL and monitored by further 21 days. After euthanasia, all animals were autopsied and the spleen was collected for weight and immunehistochemistry analyses. Additionally, peritoneal washing was done and a ÃƒÂ¢Ã¢â€šÂ¬Ã…â€œpoolÃƒÂ¢Ã¢â€šÂ¬Ã‚Â of samples from each group was prepared to be analyzed by flow citometry. Results: Mice treated with biotherapic 30x and thymulin 5cH showed similar profile, different from controls, in which a switch of lymphocytes/phagocytes proportion in the peritoneum was seen, followed by predominance of B1b cells in relation to conventional B and T cells (X2, p=0.005). Regarding to T cell population, in the contrary to control, CD4+ cells were predominant in relation to CD8+ cells (X2, p=0.0001). The immunehistochemistry revealed increase in the number of activated CD11b+ macrophages in spleen (p<0.05), but no difference were detected in the spleen lymphocytes profile. Conclusion: The results show that the action of biotherapic 30x and thymulin 5cH have similar immune modulation effects, improving the innate immune response.
Copyright (c) 2021 Camila Siqueira, Leoni Bonamin, Priscila Motta, Thayná Cardoso, Cideli Coelho, JosÃƒÆ’Ã‚Â© Nelson Couceiro, Carla Holandino
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