Gene expression of mouse lung CD11b+ dendritic cells before and after ex vivo culture

Conventional dendritic cells (cDCs) in the lung drive effector T cell differentiation, and initiate allergic responses upon inhalation of allergens. However, CD11b+ cDCs are composed Ly-6C+ and Ly-6C– subpopulations, and the specific function of each subpopulation has been elusive. Since Ly-6C+ CD11b+ cDCs lost the surface display of Ly-6C upon ex vivo culture, we hypothesized that Ly-6C+ cDCs are precursors of Ly-6C– cDCs. To determine whether 2 CD11b+ cDC subpopulations become the same cell types after maturation, we compared their transcriptomes after ex vivo cultue. As controls, we also compared transcriptomes of freshly isolated Ly-6C+ and Ly-6C– cDCs. Their transcriptomes were also compared with another CD103+ cDCs and monocytes. C57BL/6 mice were inhaled house dust extract together with ovalbumin. Ly-6C+CD11b+, Ly-6C– CD11b+, and CD103+ cDCs and monocytes were purified by flow cytometric sorting. Purified lung cDCs and monocytes were cultured for 2 days. mRNA expression isolated from freshly isolated or cultured cellswas examined by the NanoString platform.