Banca de DEFESA: JOAO PEDRO MONTEIRO CAVALCANTE

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : JOAO PEDRO MONTEIRO CAVALCANTE
DATE: 28/06/2022
TIME: 14:00
LOCAL: Sala 206 do PPGCF
TITLE:

IMMUNOMODULATORY ACTIVITY OF APITOXIN AND MELIT IN MACROPHAGES ACTIVATED BY LIPOPOLYSACCHARIDE IN VITRO

KEY WORDS:

Bee venom, Apitoxin, Melittin, Immunomodulation and Murine macrophages

PAGES: 54
BIG AREA: Ciências Biológicas
AREA: Imunologia
SUBÁREA: Imunologia Aplicada
SUMMARY:

Resumo em inglês: Apitoxin is a substance produced by bees that has numerous biological activities, being melittin the main biologically active peptide. Among the immune response cells, macrophages stand out, which can act in pro- or anti-inflammatory responses, being important target cells for the search for substances capable of modulating the immune response. Thus, the aim of this study was to evaluate the immunomodulatory activity of apitoxin (API) and melittin (MEL) in macrophages activated by lipopolysaccharide (LPS) in vitro. For this, murine macrophages (J774.A1 cell line) were initially incubated for 48h with API or MEL to determine the maximum non-toxic concentration (CMNT) by using cell viability assays, whose results were 3.125 μg/mL and 0.625 μg/mL, respectively. To evaluate the immunomodulatory activity of the compounds, macrophages were activated in vitro with LPS (200 ng/mL) and treated with API or MEL. The immunomodulatory activity was investigated by flow cytometry immunophenotyping, with the evaluation of the expression of markers F4/80 and CD86 and parameters of relative size (FSC) and complexity/granularity (SSC). The production of pro- and anti- inflammatory cytokines was also evaluated using the Cytometric Bead Array (CBA) methodology. It was possible to observe that MEL decreased the expression of CD86, as well as the size and complexity/granularity of activated macrophages, while API increased the expression of CD86 and F4/80, and increased the parameters of size and relative cellular complexity. Furthermore, it was also detected that MEL decreased the percentage of intracellular TNF-α producing cells and the levels of IL-6 and TNF-
α cytokines in the cell culture supernatant. On the other hand, apitoxin increased the percentage of intracellular TNF-α producing cells, as well as the levels of IL-6 and TNF- α in the supernatant. There was no statistically significant difference in IL- 10 levels in both treatments, with API or MEL, when compared to the untreated cells stimulated with LPS. In conclusion, this study verified the immunostimulatory activity of API and the immunosuppressive/anti-inflammatory activity of MEL in macrophages in an inflammatory context stimulated by LPS in vitro thus contributing to the development of new bioactive agents with immunomodulatory activity.

 

BANKING MEMBERS:
Interno - 3182336 - EDEILDO FERREIRA DA SILVA JUNIOR
Interno - 1612086 - ENIO JOSE BASSI
Externa ao Programa - 1790572 - REGIANNE UMEKO KAMIYA