Using cutaneous leishmaniasis of mice, the existence of so-called T helper (Th) cells type 1 and type 2 had been identified more than 20 years ago. Nowadays, it is well accepted that additional T cell populations as well as B cell-mediated immunity is required for immunity against Leishmania major. Finally, using inbred mouse strains, the relevance of genetical factors that influence anti-pathogen immunity as well as elements of the skin-immune system have been identified. This protocol describes a model for murine experimental leishmaniasis that tries to mimic natural parasite transmission by several means: (1) utilization of only infectious-stage parasites that are found in sand fly saliva, (2) intradermal inoculation, and (3) infection with only 1,000 parasites similar to the numbers inoculated by an infected sand fly.
Tissue macrophages and inflammatory neutrophils represent important cells of the innate immune system responsible for various important tasks, i.e., elimination of pathogens and/or granuloma formation. Isolation of large numbers of primary phagocytes is vital for research with these cells. Within this protocol, we present a strategy for isolation of large numbers of inflammatory neutrophils and macrophages from murine skin that allows for follow-up in vivo or in vitro studies.
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