Prolonged axenic cultivation and interaction with MDCK cells on pathogenicity properties of a clinical isolate of Acanthamoeba.
Name: GUILHERME PINHO DO PRADO
Publication date: 21/07/2017
Advisor:
Name |
Role |
|---|---|
| ALOÍSIO FALQUETO | Advisor * |
Examining board:
| Name |
Role |
|---|---|
| ALOÍSIO FALQUETO | Advisor * |
| BLIMA FUX | Internal Examiner * |
Summary: The free-living amoeba (FLA) of the genus Acanthamoeba are protozoan found in all environments of the world, and can cause diseases such as Keratitis and Granulomatous Encephalitis. Advances do not know the amoebae and their invasive process, their phenotypic and genetic variability, as well as the development of diagnostic and therapeutic methodologies, are only being fulfilled with the microorganisms cultivation tool. However, their prolonged cultivation and their passage into hosts (or cells) can change a cellular machinery and modify mechanisms related to pathogenicity, making them amoebas more or less virulent. In the present work, we investigated in vitro characteristics associated with the pathogenicity of Acanthamoeba castetellanii (Genotype T4), using four samples from a clinical isolate, originating from corneal scraping, in order to investigate its virulence. Epithelial cells of the MDCK lineage lengths for checking the cytopathic effect. It was also evaluated the environment WHERE as amebas were cultured (conditioned medium) to verify the activity of proteases in gel and the cytotoxic effect generated in cultures of MDCK. The percentages of binding and response of the amoeba culture from chlorhexidine exposure were verified. The data obtained in the work confirm the pathogenic potential of the samples, especially those that underwent passage in MDCK. The environment did not produce a significant cytotoxic effect on a cell monolayer. The zimographic profile showed a very similar pattern in all the samples tested, differentiating only one sample from a long period of axenization, with a band of approximately 133 kDa increasing. It was verified that the binding percentage of the newly axenized samples was higher than the development period, and that the latter test obtains higher binding rates after interchange with MDCK. The chlorhexidine test demonstrated that, despite the sample from the same source, as sample maintenance conditions led to different behaviors. In this work was observed in two years of axenic cultivation, an amoeba loses virulence, yet it recovers it after passage in cells, as an MDCK. Collaborate for studies in Acanthamoeba advance demonstrating a plastic amoeba's potential in managing its pathogenic profile.
