Comparison of the Accessory Activity of Murine Peritoneal Cavity Macrophage Derived Dendritic Cells and Peritoneal Cavity Macrophages in a Mixed Lymphocyte Reaction.

Abstract

ABSTRACT. A detailed comparison of the accessory cell activities was carried out among murine peritoneal cavity macrophages (PEC-MΦ), peritoneal cavity macrophages stimulated with granulocyte-macrophage colony stimulating factor (GM-CSF) plus interleukin 4 (IL-4), the most popular cytokine combination widely used to generate dendritic cells (DC) and peritoneal cavity macrophage-derived DC (PEC-DC) using a two-way mixed lymphocyte reaction (MLR). All the cell types used efficiently induced statistically significant naïve T cell proliferation at all culture time points and responder:stimulator ratios used. However, marked differences were noted in the magnitude of the proliferative responses. These variations may be attributed to the intensity of expression of MHC class II glycoproteins, as well as the actual numbers of MHC class II + cells. KEY WORDS: mixed lymphocyte reaction, peritoneal cavity macrophage, peritoneal cavity macrophage-derived dendritic cell.

Phenotype and Function of Murine Discrete Peyer's Patch Macrophage Derived-Dendritic Cells

“…mAbs were obtained from either Pharmingen (Pharmingen International, Becton Dickinson Company) for anti-(CD11c, CD80, CD86, CD16/CD32 and MHC class II) or Serotech (Serotec Ltd., Oxford, U.K.) for, anti-(CD34, F4/ 80 and DEC-205). Macrophage and dendritic cell generation: Single cells suspensions from DPP of mice were obtained using a modification of previously described methods [18,19,23,24]. Immediately after slaughter, the skin covering the abdomen was removed.…”

“…In parallel studies [22][23][24][25][26] have shown that Mø can differentiate into functional DC. Exposure to macrophage colony-stimulating factor (M-CSF) induces only macrophage formation, whereas granulocyte-macrophage colony-stimulating factor (GM-CSF) also induces DC formation [28,49].…”

“…Peyer's Patches (PP) are the major organized lymphoid structures involved in the induction of mucosal immune responses in the intestine [22][23][24]. Joseph Conrad Peyer in 1677 provided the first accurate description of these discrete lymphoid aggregates in the intestinal wall, which have come to bear his name [14,35].…”

“…Although the initial transformation into a DC form is reversible, exposure to pro-inflammatory cytokines such as tumor necrosis factor-α (TNFα) induces an apparently irreversible maturation to DC [28,49]. GM-CSF, TNFα, and interleukin-4 are widely used to induce DC development from blood monocytes, and this is currently the favoured procedure for generating DC for immunotherapy [19,[22][23][24][25][26]49]. Since DC constitute about 1% of the total body cell population, they are very difficult to isolate.…”

“…The strategic location of DPP along the intestinal wall and the rich presence of cell types involved in the induction of mucosal immune responses coupled with the need to isolate or generate DC in large numbers for in vivo and in vitro immunological responses, analysis of DC function and molecular mechanisms involved in antigen processing and presentation prompted us to investigate the suitability of using DPP macrophages as DC progenitors. Thus the present study was aimed at developing a protocol for the isolation of a functional murine discrete Peyer's patch macrophage-derived DC population using growth factors, as well as studying the effects of early cytokines IL-4 and GM-CSF that have been shown to stimulate the proliferation of myeloid progenitors into DC in our previous work [22][23][24][25][26] and having generated the DC, characterize them using three well established criteria: morphology; phenotype; and function.…”