Heparin Induces Differentiation of CD1a+ Dendritic Cells from Monocytes: Phenotypic and Functional Characterization

Abstract

Dendritic cells (DCs) play important roles in initiation and regulation of immune responses. DCs derived from human monocytes can be classified according to presence of CD1a molecules. Although CD1a+ DCs can be prepared from monocytes in media containing GM-CSF, IL-4, and FCS, it has been reported that CD1a+ DCs could not be easily obtained from monocytes using media containing human serum or plasma. In this study, we demonstrate for the first time that heparin can reliably induce differentiation of CD1a+ DCs from monocytes with or without autologous serum or plasma. The development of CD1a+ DCs is heparin concentration dependent (0–50 U/ml). Comparing with CD1a− DCs developed without heparin, CD1a+ DCs express higher CD40 and CD80 and lower CD86. Both CD1a+ and CD1a− DCs express similar levels of HLA-DR. CD80, CD86, HLA-DR, and CD40 are proportionally up-regulated when both types of DCs are stimulated with LPS or LPS plus IFN-γ. The effect of heparin is neutralized by heparin-binding proteins, such as protamine sulfate, platelet factor-4, and β-thromboglobulin. Functionally, heparin-treated DCs respond to LPS or LPS plus IFN-γ with higher IL-10 and less IL-12 production than heparin-untreated DCs. Heparin-treated DCs are more potent in priming allogeneic and autologous CD4+ T cells to proliferate and to produce both type 1 and type 2 cytokines. The results of our study show that CD1a+ DCs can be prepared from monocytes ex vivo without using xenogeneic serum and may be used for immunotherapy.

Small interference RNA modulation of IL‐10 in human monocyte‐derived dendritic cells enhances the Th1 response

“…This finding underscores the basis for the ineffectiveness of anti-IL-10 Ab to enhance Th1 induction via binding and neutralization of secreted IL-10. A recent study suggested the reverse relationship between the production of IL-10 and IL-12 in activated DC [30], a finding also observed in the current study. Neutralization of the secreted IL-10 by Ab and IL-10 antisense oligodNTP could enhance the production of IL-12 [31].…”

Glycosaminoglycans are a potential cause of rheumatoid arthritis

“…Cellular Effects. GAGs display complex biological activities toward various cells, including lymphocytes, monocytes, dendritic, and stromal cells (10,17,(20)(21)(22)(23). We examined cellular responses to GAGs to identify a correlation with disease development in mice.…”

Intranodal injection of semimature monocyte-derived dendritic cells induces T helper type 1 responses to protein neoantigen

“…40,41 Our monocyte-derived DCs generated in culture with GM-CSF and IL-4 in the presence of FCS were CD1a ϩ and showed a typical phenotype of intermediate maturity expressing low levels of CD83. 42,43 In addition, these DCs showed a strong tendency for further maturation as shown by the rapid increase in CD83 ϩ DCs after a 4-hour pulsing with KLH. Lipopolysaccharides (LPS), which are notably present in FCS, may contribute, through the engagement of pattern recognition receptor TLR-4, to drive resting monocyte-derived DCs into the DC1 development pathway.…”