Human XCR1+ Dendritic Cells Derived In Vitro from CD34+ Progenitors Closely Resemble Blood Dendritic Cells, Including Their Adjuvant Responsiveness, Contrary to Monocyte-Derived Dendritic Cells

Abstract

Human monocyte-derived dendritic cell (MoDC) have been used in the clinic with moderately encouraging results. Mouse XCR1+ DC excel at cross-presentation, can be targeted in vivo to induce protective immunity, and share characteristics with XCR1+ human DC. Assessment of the immunoactivation potential of XCR1+ human DC is hindered by their paucity in vivo and by their lack of a well-defined in vitro counterpart. We report in this study a protocol generating both XCR1+ and XCR1− human DC in CD34+ progenitor cultures (CD34-DC). Gene expression profiling, phenotypic characterization, and functional studies demonstrated that XCR1− CD34-DC are similar to canonical MoDC, whereas XCR1+ CD34-DC resemble XCR1+ blood DC (bDC). XCR1+ DC were strongly activated by polyinosinic-polycytidylic acid but not LPS, and conversely for MoDC. XCR1+ DC and MoDC expressed strikingly different patterns of molecules involved in inflammation and in cross-talk with NK or T cells. XCR1+ CD34-DC but not MoDC efficiently cross-presented a cell-associated Ag upon stimulation by polyinosinic-polycytidylic acid or R848, likewise to what was reported for XCR1+ bDC. Hence, it is feasible to generate high numbers of bona fide XCR1+ human DC in vitro as a model to decipher the functions of XCR1+ bDC and as a potential source of XCR1+ DC for clinical use.

GM-CSF Mouse Bone Marrow Cultures Comprise a Heterogeneous Population of CD11c+MHCII+ Macrophages and Dendritic Cells

“…Accordingly, GM-DCs might constitute a useful in vitro model to study the properties of migratory DCs. To evaluate the relationship between GM-DCs and the two major classical DC subsets, we analyzed the expression of genes specific to the CD8a + and CD8a À DC families (Balan et al, 2014;Mayer et al, 2014). GM-DCs did not appear to belong to either DC subset ( Figure S6).…”

Dynamics and Transcriptomics of Skin Dendritic Cells and Macrophages in an Imiquimod-Induced, Biphasic Mouse Model of Psoriasis

“…The chips were washed and stained in the GeneChip Fluidics Station 450 (Affymetrix) and scanned with the GeneChip Scanner 3000 7G (Affymetrix) at a resolution of 0.7 mm. Microarray analyses were performed as previously described (18)(19)(20). Hierarchical clustering and principal component analysis (PCA) were performed on the 5558 probes showing a fold change $2 and a p value ,0.05 for at least one cell type between day 5 or 11 versus day 0 using the LIMMA statistical package of the R program.…”

“…The most impressive enrichment observed in GSEA corresponded to gene sets associated with cell proliferation and denoted as "CCNB2" and "Cell cycle" (20) (Supplemental Fig. 2F).…”

CD1c+ blood dendritic cells have Langerhans cell potential

“…25 Although the cultures contained serum, as in other recent publications describing DC differentiation, 24,26 the factors GM-CSF, TGFb, and BMP7 are expressed by epithelial tissues under physiologic conditions. 23 Previous experiments on CD34 Finally, these data explain the puzzling claim that there are CD1a…”