Limits...
Synovial fluid myeloid dendritic cells display important differences compared to monocyte-derived dendritic cells prepared in vitro.

Moghaddami M, James M, Whittle SL, Cleland LG - Clin Transl Immunology (2014)

Bottom Line: LPS reduced 1,25D-induced expression of PGDS in MDDCs.SFDCs and MDDCs display similar basal characteristics but differ in PGDS expression and responsiveness to LPS and 1,25D.MDDCs have limitations as a model of SFDCs which have differentiated in vivo.

View Article: PubMed Central - PubMed

Affiliation: Arthritis Research Laboratory, Hanson Institute, SA Pathology , Adelaide, South Australia , Australia ; Department of Medicine, University of Adelaide , Adelaide, South Australia, Australia.

ABSTRACT
The object of this study was to characterise synovial fluid dendritic cells (SFDCs) with regard to morphology, phenotype and responses to 1,25hydroxy-cholecalciferol (1,25D) and lipopolysaccharide (LPS), and to compare these characteristics with those of peripheral blood (PB) monocyte-derived DCs (MDDCs). SF was aspirated from knees with inflammatory effusions. PB samples were obtained contemporaneously. SFDCs were separated by flow cytometry. Morphology was determined on cytosmears. Expression of accessory molecules, cytokines and prostaglandin synthases mRNA was quantified by reverse transcription PCR. Analyses were performed on freshly prepared DCs and after incubation with 1,25D and LPS, separately and in combination. SFDCs and MDDCs displayed broadly similar morphology. Expression of accessory molecules, cytokines, cyclooxygenase-2 (COX-2) and prostaglandin E-synthase (PGES) was similar. SFDCs, but not MDDCs, expressed prostaglandin D-synthase (PGDS). PGDS was lost on incubation with SFDCs, but was induced by 1,25D in MDDCs. LPS in the presence or absence of 1,25D, induced interleukin 23 (IL23), IL1β and tumour necrosis factor-α in SFDCs and MDDCs, with SFDC showing stronger expression of these cytokines. 1,25D in combination with LPS induced PGES and enhanced LPS induction of IL6 in SFDCs and MDDCs. LPS reduced 1,25D-induced expression of PGDS in MDDCs. SFDCs and MDDCs display similar basal characteristics but differ in PGDS expression and responsiveness to LPS and 1,25D. MDDCs have limitations as a model of SFDCs which have differentiated in vivo.

No MeSH data available.


Related in: MedlinePlus

Four colour flow cytometric analysis of cells prepared from SF aspirates. (a) the selected gate based on forward and side scatter of light; (b) selection of HLADR+ and CD11b+ cells; (c) shows majority of HLADR+ CD11b+CD11c+ cells are CD14+ monocytes and 15% are CD14− myeloid DCs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4232072&req=5

fig1: Four colour flow cytometric analysis of cells prepared from SF aspirates. (a) the selected gate based on forward and side scatter of light; (b) selection of HLADR+ and CD11b+ cells; (c) shows majority of HLADR+ CD11b+CD11c+ cells are CD14+ monocytes and 15% are CD14− myeloid DCs.

Mentions: SFDCs freshly prepared using flow cytometry (Figure 1) and MDDCs both displayed the cytoplasmic and cell surface morphology of immature DCs, that is, medium size with smooth or ruffled borders (Figures 2a and b). However, the feature of eccentric location of the nucleus, which is regarded as a typical feature of immature DCs, was seen in MDDCs (Figure 2b), whereas the indented nucleus was often more centrally located in many SFDCs (Figure 2a). Culture of both SFDCs and MDDCs with the TLR-4 ligand LPS yielded the morphology characteristic of mature DCs (that is, variably located nuclei and veiled cytoplasm; Figures 2c and d).


Synovial fluid myeloid dendritic cells display important differences compared to monocyte-derived dendritic cells prepared in vitro.

Moghaddami M, James M, Whittle SL, Cleland LG - Clin Transl Immunology (2014)

Four colour flow cytometric analysis of cells prepared from SF aspirates. (a) the selected gate based on forward and side scatter of light; (b) selection of HLADR+ and CD11b+ cells; (c) shows majority of HLADR+ CD11b+CD11c+ cells are CD14+ monocytes and 15% are CD14− myeloid DCs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232072&req=5

fig1: Four colour flow cytometric analysis of cells prepared from SF aspirates. (a) the selected gate based on forward and side scatter of light; (b) selection of HLADR+ and CD11b+ cells; (c) shows majority of HLADR+ CD11b+CD11c+ cells are CD14+ monocytes and 15% are CD14− myeloid DCs.
Mentions: SFDCs freshly prepared using flow cytometry (Figure 1) and MDDCs both displayed the cytoplasmic and cell surface morphology of immature DCs, that is, medium size with smooth or ruffled borders (Figures 2a and b). However, the feature of eccentric location of the nucleus, which is regarded as a typical feature of immature DCs, was seen in MDDCs (Figure 2b), whereas the indented nucleus was often more centrally located in many SFDCs (Figure 2a). Culture of both SFDCs and MDDCs with the TLR-4 ligand LPS yielded the morphology characteristic of mature DCs (that is, variably located nuclei and veiled cytoplasm; Figures 2c and d).

Bottom Line: LPS reduced 1,25D-induced expression of PGDS in MDDCs.SFDCs and MDDCs display similar basal characteristics but differ in PGDS expression and responsiveness to LPS and 1,25D.MDDCs have limitations as a model of SFDCs which have differentiated in vivo.

View Article: PubMed Central - PubMed

Affiliation: Arthritis Research Laboratory, Hanson Institute, SA Pathology , Adelaide, South Australia , Australia ; Department of Medicine, University of Adelaide , Adelaide, South Australia, Australia.

ABSTRACT
The object of this study was to characterise synovial fluid dendritic cells (SFDCs) with regard to morphology, phenotype and responses to 1,25hydroxy-cholecalciferol (1,25D) and lipopolysaccharide (LPS), and to compare these characteristics with those of peripheral blood (PB) monocyte-derived DCs (MDDCs). SF was aspirated from knees with inflammatory effusions. PB samples were obtained contemporaneously. SFDCs were separated by flow cytometry. Morphology was determined on cytosmears. Expression of accessory molecules, cytokines and prostaglandin synthases mRNA was quantified by reverse transcription PCR. Analyses were performed on freshly prepared DCs and after incubation with 1,25D and LPS, separately and in combination. SFDCs and MDDCs displayed broadly similar morphology. Expression of accessory molecules, cytokines, cyclooxygenase-2 (COX-2) and prostaglandin E-synthase (PGES) was similar. SFDCs, but not MDDCs, expressed prostaglandin D-synthase (PGDS). PGDS was lost on incubation with SFDCs, but was induced by 1,25D in MDDCs. LPS in the presence or absence of 1,25D, induced interleukin 23 (IL23), IL1β and tumour necrosis factor-α in SFDCs and MDDCs, with SFDC showing stronger expression of these cytokines. 1,25D in combination with LPS induced PGES and enhanced LPS induction of IL6 in SFDCs and MDDCs. LPS reduced 1,25D-induced expression of PGDS in MDDCs. SFDCs and MDDCs display similar basal characteristics but differ in PGDS expression and responsiveness to LPS and 1,25D. MDDCs have limitations as a model of SFDCs which have differentiated in vivo.

No MeSH data available.


Related in: MedlinePlus