Obtaining cells from colon of dog with leishmaniasis for flow cytometric analysis
In dogs, a parasite burden is evident throughout the gastrointestinal tract but not is observed pathological alterations. Our aim was isolating lamina propria cells from dogs infected with leishmaniasis to further investigate if the parasite in the gut is correlated with the pathogenesis of the visceral infection. The biopsies were obtained of dogs and incubated with collagenase II. In the cells suspension was added antibodies against superficial and intracellular markers. Few studies of the markers used in this work have been done in the lamina propria cells in leishmaniasis. It is of great importance to determine markers in this tissue especially those who are described as being involved with the persistence of Leishmania infection within cells due to the fact that the infection in the gut does not induce lesions. Furthermore, work is ongoing in terms of investigating other segments of the GIT, a systematic study, from dogs with Leishmania
A pdf of the full version of this protocol can be found "here":http://www.nature.com/protocolexchange/system/uploads/2040/original/Fullprotocol-_protex.2011.273.pdf?1323950759
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Full Protocol Obtaining cells from colon of dog with leishmaniasis for flow cytometric analysis
Posted 15 Dec, 2011
Obtaining cells from colon of dog with leishmaniasis for flow cytometric analysis
Posted 15 Dec, 2011
In dogs, a parasite burden is evident throughout the gastrointestinal tract but not is observed pathological alterations. Our aim was isolating lamina propria cells from dogs infected with leishmaniasis to further investigate if the parasite in the gut is correlated with the pathogenesis of the visceral infection. The biopsies were obtained of dogs and incubated with collagenase II. In the cells suspension was added antibodies against superficial and intracellular markers. Few studies of the markers used in this work have been done in the lamina propria cells in leishmaniasis. It is of great importance to determine markers in this tissue especially those who are described as being involved with the persistence of Leishmania infection within cells due to the fact that the infection in the gut does not induce lesions. Furthermore, work is ongoing in terms of investigating other segments of the GIT, a systematic study, from dogs with Leishmania
A pdf of the full version of this protocol can be found "here":http://www.nature.com/protocolexchange/system/uploads/2040/original/Fullprotocol-_protex.2011.273.pdf?1323950759
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