Evaluation of immune response of BALB/c to homeopathic solutions

Abstract

Introduction: 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) – administration of thymulin 5cH, Group B (5 animals) – administration of biotherapic 30x, Group C (5 animals) – 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 g/200L 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 “pool” 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