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Phenotypic and Functional Characterization of Mouse Hepatic CD8⁺ Lymphoid-Related Dendritic Cells¹

Peta J. O’Connell^*, Adrian E. Morelli^*, Alison J. Logar^* and Angus W. Thomson^2,*,

^* The Thomas E. Starzl Transplantation Institute and Department of Surgery, and Department of Molecular Genetics and Biochemistry, University of Pittsburgh, Pittsburgh, PA 15213

Recently, attention has focussed on phenotypic and functional differences between classic myeloid dendritic cells (DC), and DC that reportedly develop from an early, committed lymphoid precursor. In mice, DC from these separate hemopoietic lineages differ by their surface expression of CD8. We undertook a comparative study of CD8⁺ (CD11b^low; lymphoid-related) and CD8^- (CD11b^high; myeloid) DC isolated from mouse liver. CD8⁺ and CD8^- DC each constituted 1.0% of the freshly isolated, normal nonparenchymal cells (NPC). Both populations were enriched 10–15% by overnight culture and metrizamide density centrifugation. Flt3 ligand (Flt3L) potently induced equal expansion of both subsets in vivo. Tissue-resident CD8⁺ DC, freshly isolated from Flt3L-treated mice, existed primarily as immature cells (CD11c⁺, CD11b^low, CD40^-/low, CD80^low, CD86^low, MHC class II^low), consistent with previous observations regarding bulk DC freshly isolated from nonlymphoid tissues. Following overnight culture in GM-CSF, CD8⁺ DC underwent phenotypic and functional maturation equivalent to that observed for CD8^- DC. CD95 ligand (FasL) mRNA was detected in both immature and mature DC of each subset. In vitro analysis confirmed that flow-sorted, mature CD8⁺ and CD8^- DC were strong and equally efficient stimulators of allogeneic T cell proliferation in primary MLR. Both immunohistochemical and genomic DNA analysis revealed that in vivo, sorted CD8⁺ DC trafficked from s.c. sites to T cell areas of allogeneic lymphoid tissue and were equally efficient at priming naive T cells compared with CD8^- DC. This is the first comparative study of lymphoid-related DC isolated from nonlymphoid tissue.

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