A method for the isolation and analysis of leucocytic cells from Leishmanial ear lesions in mice

The standard model of experimental cutaneous leishmaniasis involves infection of mice with Leishmania major in a single footpad or the rump, and analysis of the subsequent immune response in draining lymph nodes. Relatively few studies have examined the lesion directly. Here, we describe a method for the isolation of cells from established leishmanial lesions in mouse ears. After physical disruption of lesion tissue and isolation of cells on density gradients, a variety of leucocytic cell phenotypes were identified by flow cytometry and cytology. The phenotypes of the viable cells obtained were similar, in proportion, to those observed in histologic sections of ear lesions. This technique may be useful for studying lesion-specific cell function within the first weeks after infection with Leishmania parasites. (C) 1999 Elsevier Science B.V. All rights reserved.