Differentiation of blood monocytes to dendritic cells following reverse transmigration across the blood-brain barrier (94.6)

Abstract

Abstract Dendritic cells (DC) are absent in the normal human central nervous system (CNS) but their presence has been documented in neuroinflammation. The origin of CNS DC is still controversial. The objective of this study was to determine whether monocytes that cross the blood-brain barrier (BBB) differentiate into DC. Primary human brain microvessel endothelial cells (HBMEC) were cultured on collagen gels. Purified human CD14+ monocytes were incubated with confluent HBMEC for 48h and the reverse transmigrated (RT) cells were characterized by FACS analysis. Approximately 50% of the monocytes reverse transmigrated across HBMEC monolayers and 70% of the RT cells expressed DC-SIGN. Monocytes incubated on gels in the absence of HBMEC retained their phenotype. The percentage of RT cells expressing CD83 increased from 25% to 70% when human CNS myelin was incorporated into the gels, and there was significant increase in CCR7 expression. Analysis of culture supernatants showed high levels of IL-6, TNF-α, PGE2 and IL-1β when HBMEC were cultured on myelin gels suggesting that these cytokines may contribute to the differentiation of monocytes to mature DC. There was an associated increase of MIP-3β, MCP-1, MIP-1α, and MIP-1β in the supernatants. Mixed lymphocyte reactions by co-culturing CFSE-labeled T cells and RT cells showed that RT cells were capable of stimulating allogeneic T cell proliferation. In conclusion, the results suggest that monocytes which reverse-transmigrate across HBMEC differentiate into mostly mature DC capable of inducing T cell proliferative responses. (Supported by MSS, Canada)