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Doctoral Thesis
Full name
Narciso Junior Vieira
Knowledge Area
Date of Defense
São Paulo, 2016
Mendes, Joao Gustavo Pessini Amarante (President)
Bargieri, Bruna Cunha de Alencar
Câmara, Niels Olsen Saraiva
Denapoli, Priscila Martins Andrade
Ferrari, Merari de Fatima Ramires
Title in Portuguese
Papel da RAB2A, RAB5A, RAB17 e RAB18 na função efetora de células citotóxicas.
Keywords in Portuguese
Células citotóxicas
Células natural killer
Tráfego intracelular
Abstract in Portuguese
Linfócitos T CD8 e células NK atuam no combate à infecções por bactérias intracelulares, vírus e células tumorais, provocando a morte dessas células por meio da secreção de grânulos citotóxicos. Proteínas RAB GTPase têm se destacado em estudos de tráfego intracelular, porém, são escassos dados sobre o papel destas proteínas em células citóxicas. Um estudo prospectivo de proteômica realizado por nosso grupo identificou a RAB2A, RAB5A, RAB17 e RAB18 em grânulos citotóxicos. Análises mais aprofundadas revelaram que a RAB2A está associada a proteínas como LAMP-1 e LAMP-2, enquanto que RAB5A, RAB17 e RAB18 estavam presentes na mesma linhagem em um contexto não contemplado neste estudo. Desenvolvemos ainda uma abordagem de silenciamento gênico da RAB2A, e por fim, adaptamos uma série de protocolos de simples execução e baixo custo para avaliar funções efetoras de células NK. O conhecimento da maquinaria secretória é fundamental, uma vez que defeitos nas vias de tráfego intracelular constituem a base de um grande número de doenças que desencadeiam quadros fatais.
Title in English
Role of RAB2A, RAB5A, RAB17 andRAB18 in effector functions of cytotoxic cells.
Keywords in English
Cytotoxic cells
Intracellular trafficking
Natural killer cells
Abstract in English
CD8 T lymphocytes and NK cells fight against infections by intracellular bacteria, viruses and tumor cells by killing those cells through the secretion of cytotoxic granules. RAB GTPase has been highlighted in studies of intracellular trafficking, however there are scarce reports regarding the role of these proteins in cytotoxic cells. A proteomic study performed by our group identified RAB2A, RAB5A, RAB17 and RAB18 in cytotoxic granules. Further analysis revealed that RAB2A is associated with LAMP-1 and LAMP-2, while RAB5A, RAB17 and RAB18 were present in the same cell line, but in a context not included in this study. We also have developed a gene silencing approach for RAB2A and adapted a number of protocols, simple and low-cost, that can be used to evaluate effector functions of natural killer cells The knowledge of secretory machinery involved in the movement cytotoxic granules of cytotoxic cells is critical, since defects in intracellular trafficking pathways constitute the basis for a large number of diseases which trigger death.
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  • CLEMENTE, T., et al. In vivo assessment of specific cytotoxic T lymphocyte killing [doi:10.1016/j.ymeth.2013.02.007]. Methods (San Diego, Calif., Print) [online], 2013, p. 105-109.
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