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Characterization of tick cell lines and their use as a substrate for the growth, maintenance and protein expression profile of Anaplasma marginale


Ticks are arthropods of great medical and veterinary importance, with many species responsible for transmitting viruses, bacteria and protozoa to their hosts. Some of the tick-borne pathogens are difficult to grow in vitro, and do not grow in artificial media. Tick cell culture has been an excellent tool to isolate and study pathogens, allowing the production of antigenic material for diagnostic tests, production of antibodies and vaccines, and for studies of host-vector-pathogen relationships. In this sense, this study aims to characterize tick cell lines and use them as a substrate for the growth, maintenance and protein expression profile of Anaplasma marginale. For this, we will use primary cultures and established lineages of tick cells from Amblyomma sculptum (Berlese) and Rhipicephalus (Boophilus) microplus (Canestrini) eggs. The primary cultures will be prepared with egg masses and kept at 30 ° C in L-15B medium (Leibovitz) supplemented with Bovine Fetal Serum, subcultured when they reach the necessary confluence and cryopreserved in several passages. For the characterization of cells in culture, morphological characterization with histological techniques and karyotyping will be performed. The cells inoculated with Anaplasma marginale canis will be analyzed by slide preparations and counts of infected cells under optical microscopy, by real-time PCR and by Transmission Electron Microscopy. In addition, a differential proteomic analysis of infected and uninfected cells will be carried out, aiming at the search for proteins that may be used as vaccine antigens in the future. (AU)

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