In vitro T lymphocyte/NK cell cytotoxicity against enhanced green fluorescent protein expressing targets (2005)
- Authors:
- Autor USP: MENDES, JOAO GUSTAVO PESSINI AMARANTE - ICB
- Unidade: ICB
- Assunto: IMUNOLOGIA
- Language: Inglês
- Abstract: Introduction and Objectives: Cell-mediated cytotoxicity plays an important role in immune surveillance to tumors and is also responsible for destruction of allografts in vivo. The major cells involved in this process are cytotoxic T lymphocytes and Natural Killer (NK) cells. Our aim was to study cell-mediated cytotoxicity against multiple targets including NK- or CTLsensitive cells expressing EGFP and comparing the results with other protocols. Methods and Results: Non-adherent lymphocytes were used as effector cells in cytotoxic assays. Two different approaches were used to discriminate effectors cells from targets: labeling the latter ones with DiOC18 or using EGFP-expressing targets cells. First of all, we developed the following EGFP-expressing cells: a) L929, a fibroblast cell line derived from C3H/An mouse; b) B16F10, a NK-sensitive melanoma derived from C57Bl/6; c) RMA/RMAS, which are derived from C57Bl/6 lymphoma T cell and are NK-insensitive and -sensitive targets, respectively, and d) K562, a human erythroleukemia cell line. Best results were obtained after 16 hours of incubation for cytotoxic assays, using 2 microM DiOC18 at 40:1 ratio. L929, K562 and YAC-1, showed higher fluorescence which is important to our aim, since higher gain of fluorescence allows better discrimination between effector and target cells. On the other hand, incubation with digitonin to simulate necrosis does not interfere with DiOC18 staining, but results in lower fluorescence ofthe EGFP-positive cells. As expected the killing was greater against L929 (80%) than B16F10 (12%) possibly due to an allogeneic response. Conclusion: We showed that according to the target cell line different methods should be addressed. We also showed that EGFP-expressing targets are a useful tool to study cellmediated cytotoxicity in vitro. Finally, this tool could be used for in vivo studies to understand antitumoral cytotoxicity as well as graft rejection
- Imprenta:
- Publisher: Comissão de Cultura e Extensão Universitária do ICB/USP
- Publisher place: São Paulo
- Date published: 2005
- Source:
- Título: Resumos
- Conference titles: Congresso do Instituto de Ciências Biomédicas
-
ABNT
CASTRO, Monica Alexandra Yon e AMARANTE-MENDES, João Gustavo Pessini. In vitro T lymphocyte/NK cell cytotoxicity against enhanced green fluorescent protein expressing targets. 2005, Anais.. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP, 2005. . Acesso em: 28 dez. 2025. -
APA
Castro, M. A. Y., & Amarante-Mendes, J. G. P. (2005). In vitro T lymphocyte/NK cell cytotoxicity against enhanced green fluorescent protein expressing targets. In Resumos. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP. -
NLM
Castro MAY, Amarante-Mendes JGP. In vitro T lymphocyte/NK cell cytotoxicity against enhanced green fluorescent protein expressing targets. Resumos. 2005 ;[citado 2025 dez. 28 ] -
Vancouver
Castro MAY, Amarante-Mendes JGP. In vitro T lymphocyte/NK cell cytotoxicity against enhanced green fluorescent protein expressing targets. Resumos. 2005 ;[citado 2025 dez. 28 ] - Modulation of fasl and activation-induced cell death (aicd) of do11.10 T-cells hybridoma by imiquimod
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