Sorting of Leishmania-bearing dendritic cells reveals subtle parasite-induced modulation of host-cell gene expression
- Lecoeur, Hervé
- de La Llave, Emilie
- Osorio y Fortéa, José
- Goyard, Sophie
- Kiefer-Biasizzo, Helene
- Balazuc, Anne-Marie
- Milon, Geneviève
- Prina, Eric
- Lang, Thierry
DOIAbstract
International audienceOnce in the mouse skin, Leishmania (L) amazonensis amastigotes are hosted by professional mononuclear phagocytes such as dendritic cells (DCs). When monitored after parasite inoculation, the frequency of amastigote-hosting DCs is very low (<1%) in both the skin and skin-draining lymph nodes. Therefore, we designed and validated an efficient procedure to purify live amastigotes-hosting DCs with the objective to facilitate quantitative and qualitative analysis of such rare cells. To this end, a L. amazonensis transgenic parasite expressing DsRed2 fluorescent protein was generated and added to mouse bone marrow-derived DC cultures. Then, a high speed sorting procedure, performed in BSL-2 containment, was setup to pick out...
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