Staining O3OD cell line and primary dog macrophages by means of immunocytochemical and immunofluorescence staining using a CD14 marker to eventually compare healthy and Leishmania infected dogs

Abstract

Canine Leishmaniosis (CanL) is caused by the zoonotic protozoan Leishmania infantum and is transmitted by the bites of female phlebotomine sandflies. Via the phlebotomine sand fly, infection can be transmitted from dogs to humans, resulting in visceral or cutaneous Leishmaniasis. Due to global warming, which favors the spread of the sand fly, and the long-distance importation of dogs, CanL is expanding to new locations in Europe. Until now it is unknown how many dogs are infected with Leishmania and other vector-borne diseases like Ehrlichiosis and Dirofilariasis. The target cell of Leishmania is the macrophage, this is where replication takes place. M1 and M2 macrophages induce a different immunological response, either beneficial or unfavorable for disease progression. Therefore, distinguishment between M1 and M2 macrophage subtypes could be important for predicting the development of Leishmaniosis. We hypothesized that there is a difference between the M1 and M2 macrophages of healthy and Leishmania infected dogs. During this study we stained O3OD cell line and primary dogs macrophages by means of respectively immunocytochemical and immunofluorescence staining, using a CD14 marker. However, the initial goal of this study to compare the macrophages staining in DNAB samples obtained from healthy and Leishmania infected dogs was not achieved. Therefore, the drafted H0 and H1 hypotheses cannot yet be answered with the findings of this study and future research is necessary. These future answers can give us more insight in the role of macrophages in the pathology of Leishmania and may contribute to proper monitoring and treatment of the disease in dogs.