Antigen presentation function of brain-derived dendriform cells depends on astrocyte help

In mouse brain primary culture, supplementation with granulocyte macrophage colony-stimulating factor (GM-CSF) induces development of dendriform cells emerging on the astroglia monolayer, As revealed by flow cytofluorimetric analysis, >70% of isolated cells are CD11c(+) and express the dendritic cell (DC) marker 33D1, Additional expression of F4/80 and CD11b suggests a myeloid origin of these cells, The lymphoid DC marker CD8 alpha is lacking while DEC-205 has been detected on similar to 10% of the cells, When freshly isolated, such brain-derived DC-like cells are excellent antigen-presenting cells (APC) but their functional capability is lost during subculture with GM-CSF. In contrast, their antigen presentation function remains stable in the presence of GM-CSF plus astrocytes or astrocyte-conditioned medium. The responsible astrocytic activity co-fractionates with macrophage colony-stimulating factor (M-CSF), Neutralization of the activity with anti-M-CSF antibody and substitution with recombinant M-CSF provide evidence that, in addition to GM-CSF, M-CSF is required to preserve the functional capability of these brain-derived APC, Responsiveness of the isolated cells to M-CSF is substantiated by the expression of c-fms/M-CSF receptor gene, Consistently, GM-CSF proves stimulatory for astrocytes by up-regulating their secretion of M-CSF. Furthermore, depletion or blocking of endogenous M-CSF in primary brain cell culture prevents the development of functionally active APC regardless of exogenous GM-CSF, In sum, these findings ascribe an immature DC phenotype to GM-CSF-grown myeloid brain cells and indicate a role for astrocytic M-CSF in maintaining their antigen presentation function.