Neutrophil-Derived CCL3 Is Essential for the Rapid Recruitment of Dendritic Cells to the Site of Leishmania major Inoculation in Resistant Mice

Abstract

Neutrophils are rapidly and massively recruited to sites of microbial infection, where they can influence the recruitment of dendritic cells. Here, we have analyzed the role of neutrophil released chemokines in the early recruitment of dendritic cells (DCs) in an experimental model of Leishmania major infection. We show in vitro, as well as during infection, that the parasite induced the expression of CCL3 selectively in neutrophils from L. major resistant mice. Neutrophil-secreted CCL3 was critical in chemotaxis of immature DCs, an effect lost upon CCL3 neutralisation. Depletion of neutrophils prior to infection, as well as pharmacological or genetic inhibition of CCL3, resulted in a significant decrease in DC recruitment at the site of parasite inoculation. Decreased DC recruitment in CCL3 ^−/− mice was corrected by the transfer of wild type neutrophils at the time of infection. The early release of CCL3 by neutrophils was further shown to have a transient impact on the development of a protective immune response. Altogether, we identified a novel role for neutrophil-secreted CCL3 in the first wave of DC recruitment to the site of infection with L. major, suggesting that the selective release of neutrophil-secreted chemokines may regulate the development of immune response to pathogens.

Author Summary

When infectious agents enter our body, neutrophils are the first cells recruited to the scene. In addition to their capacity to kill microbes, neutrophils secrete molecules that attract other cells also involved in immune defense, such as dendritic cells (DCs). Here, we investigate the secretion of DC-attracting chemokines by neutrophils after inoculation of mice with Leishmania major, a protozoan parasite that can cause long-lasting, skin ulcers in man. Following parasite inoculation, most inbred strains of mice (e.g.C57BL/6) develop self-resolving lesions, while in a few strains (e.g. BALB/c) lesions fail to heal. We report here that in “healer” C57BL/6 mice, higher numbers of DCs were attracted at the site of infection than in “non-healer” BALB/c mice. DC recruitment correlated with secretion by neutrophils of the chemokine CCL3, as indeed a markedly decreased DC recruitment was observed in C57BL/6 mice depleted of neutrophils or deprived of the capacity to produce CCL3. DC recruitment was restored upon transfer of normal neutrophils to CCL3 deficient mice. Our results reveal a crucial role for neutrophil-secreted CCL3 in early recruitment of DCs in L. major-infected “healer” mice, and suggest that the type of chemokine secreted by neutrophils will have consequences in the launching of pathogen-specific immune response.