Banca de QUALIFICAÇÃO: SUZANNE NERY SERRATH

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
STUDENT : SUZANNE NERY SERRATH
DATE: 13/12/2023
TIME: 08:00
LOCAL: Sala Virtual Remota https://bit.ly/Qualif_MSc_SuzanneNery_PGBIOEXP
TITLE:

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KEY WORDS:

Cr-LAAO, extracellular vesicle, snake venom, inflammation

PAGES: 71
BIG AREA: Ciências Biológicas
AREA: Biologia Geral
SUMMARY:

Animal venoms are an important source of proteins with high adaptability to the environment. Among these proteins is L-amino acid oxidase (LAAO), which has been studied due to its involvement in various physiopathological effects such as hemolysis, edema, hemorrhage, apoptosis induction, cytotoxicity, microbicidal activities, effects on platelets, and activation of human neutrophils. Exosomes, cellular nanovesicles, play vital roles in intercellular communication, including immune responses, and participate in some inflammatory physiopathological effects such as oxidative stress, which is the main response to neutrophil activation. Thus, this study investigates the impact of LAAO isolated from the venom of the snake Calloselasma rhodostoma (Cr-LAAO) on the release of exosomes from human neutrophils, including activation patterns, proteins identified in the vesicles, and their signaling pathways. Isolated human neutrophils were incubated with RPMI (negative control), PMA (500 ng/mL positive control), and Cr-LAAO (100 μg/mL). After 3 hours of incubation, the supernatant was collected, and exosomes were isolated by filtration followed by ultracentrifugation at 100,000 xg for 3 hours. Size characterization was performed using dynamic light scattering with the Zetasizer Nano ZS system (Malvern® Instrument). Exosome surface markers were characterized by Western Blot with the detection of CD63 and CD81. Data obtained through mass spectrometry proteomics allowed the identification of exosome proteins. Analysis of the supernatant from neutrophils stimulated by Cr-LAAO showed the presence of particles equivalent to exosomes, ranging in size from 8 to 70 nm. Western blot revealed the expression of CD63 and CD81 markers on exosomes released by neutrophils stimulated by Cr-LAAO. Proteomic analysis of exosomes identified approximately 722 proteins in all experimental groups and their interactions, and these proteins are involved in biological processes, molecular functions and cellular component. The results of this study provide a better understanding of how Cr-LAAO induces the activation of human neutrophils, leading to the release of exosomes and facilitating intercellular communication, providing important insights into possible therapeutic approaches for inflammatory and immune disorders.

COMMITTEE MEMBERS:
Externa à Instituição - BRUNA CAVECCI-MENDONÇA
Externa à Instituição - LUCIANA LYRA CASAIS E SILVA - UFBA
Presidente - ***.482.422-** - NAJLA BENEVIDES MATOS - UNIR