11/20/2022 0 Comments Rising kingdoms human 34![]() ![]() Shaded histograms, isotype controls gray line, CD1a and langerin double-negative cells black line, CD1a + langerin + cells. (C) Extended phenotype of LC-like cells (CD1c +) compared with primary Langerhans cells (Langerhans). (B) Comparison of the number of Birbeck granules per cell section between CD14 + monocytes and CD1c + DC cultures. The experiment was performed twice with 2 different donors only with GM-CSF and BMP7. Insets in the top left panel are displayed beneath and to the right. (A) EM images of Birbeck granules in CD1c + DCs cultured for 7 days showing classical “tennis racket” morphology, pentalaminar structure, and formation by endocytosis. Differences in langerin induction between monocyte subsets and pDCs for a given culture condition were not significant.įormation of Birbeck granules in cultures of CD1c + DCs. ![]() 01 compared with corresponding CD14 + monocyte culture. There were no statistically significant differences between each condition for a given subset of cells. Gating of langerin + and langerin high cells is illustrated using CD1c + DCs incubated with GM-CSF and BMP7 as an example. Mean ± SEM of 5 experiments with different donors. Lower plot: percentage of langerin high cells derived from GM-CSF+TGFβ (open bars), GM-CSF+BMP7 (gray bars), and GM-CSF+TGFβ and BMP7 (black bars) after 3 days of culture. ![]() (D) Upper plot: percentage of langerin + cells derived from GM-CSF+TGFβ (open bars), GM-CSF+BMP7 (gray bars), and GM-CSF+TGFβ and BMP7 (black bars) after 3 days of culture. Ten-thousand cells were added to each well as counted by the sorter but typically resulted in 6000 to 8000 viable cells at the start of the culture. (C) Recovery of viable cells under each condition at 7 days of culture, estimated by the total number of 4,6 diamidino-2-phenylindole (DAPI)–negative cells recorded when the culture was analyzed and run to dryness on the cytometer. ![]() (B) Time course of expression of CD1a and langerin double-positive cells showing a peak at 3 days and gradual decline in the percentage of positive cells up to 7 days of culture. Four subsets of cells were collected from 1 donor, different in each experiment. The experiment was repeated 5 times except for the panels with X-Vivo, which were repeated 3 times. (A) Sorted cells cultured for 3 days in conditions as indicated showing expression of CD1a and extracellular langerin. © 2015 by The American Society of Hematology.Įxpression of langerin and CD1a by monocytes and DCs. These data highlight a new potential precursor function of CD1c(+) DCs and demonstrate an alternative pathway of LC differentiation that may have relevance in vivo. Here we show that, although monocytes are able to express langerin, when cultured with soluble ligands granulocyte macrophage colony-stimulating factor (GM-CSF), transforming growth factor β (TGFβ), and bone morphogenetic protein 7 (BMP7), CD1c(+) dendritic cells (DCs) become much more LC-like with high langerin, Birbeck granules, EpCAM, and E-cadherin expression under the same conditions. Recent data in mice also show long-term repopulation of the LC compartment with alternative myeloid precursors. However, differentiation experiments with human lineage-negative cells and CD34(+) progenitors suggest that there is an alternative monocyte-independent pathway of LC differentiation. Human monocytes give rise to langerin-positive cells in vitro, suggesting a potential precursor role. Langerhans cells (LCs) are self-renewing in the steady state but repopulated by myeloid precursors after injury. ![]()
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