Phenotype and Function of Murine Peritoneal Cavity Macrophage Derived-Dendritic Cells.

Abstract

ABSTRACT. The accessory activity was reported in murine peritoneal cavity macrophage derived dendritic cells (PEC-DC) in a mixed lymphocyte reaction (MLR). Here we continue the characterization of the generated PEC-DC using the criteria of morphology, phenotype and other accessory function. We have demonstrated that murine peritoneal cavity macrophages can be induced to differentiate in vitro into cells exhibiting typical dendritic cell (DC) morphology, phenotype and function. The proliferative capacity of the progenitors was amplified in the first step of the culture (day 0-7) using a combination of early cytokines: interleukin 4 and granulocyte-macrophage colony-stimulating factor. The second step of the culture started at day 7 with the removal of early growth factors to allow differentiation and final maturation of DC during 2 days of culture with interferon gamma plus either Toxoplasma lysate antigen (TLA) or lipopol ysaccharide (LPS), a bacterial agent as a DC maturing agent. The resulting DC population exhibited typical DC morphology and expressed higher levels of MHC class II and the co-stimulatory molecules CD80 and CD86 compared to the untreated peritoneal cells. The generated DC cells efficiently presented soluble protein antigen to CD3 + spleen T cells.

Dendritic Cells: A Specialized Complex System of Antigen Presenting Cells.

“…Great progress has been achieved in the last decade by several groups that have succeeded in the establishment of procedures for the in vitro generation of murine and human DC from progenitors in the bone marrow, placental and umbilical cord blood, and cytokine mobilized peripheral blood [16, 38, 51, 52,79,80,91,126,151,155,156,180]. Methods have also been developed to generate DC from blood monocytes [8,112,117,119,121] and murine peritoneal cavity macrophages [88]. Long-term DC lines have also recently been generated from fetal tissues using either granulocyte macrophage colony stimulating factor (GM-CSF) [173] or stromal cell culture supernatants [157].…”

“…In our previous work we have demonstrated the effectiveness of metrizamide gradients [86,87,89]. Recent studies have shown great progress in the establishment of several procedures for the in vitro generation of DC from progenitors in the bone marrow, placental and umbilical cord blood [16,19,38,51,52,79,80,91,126,151,155,156,180]; monocytes [117,119] and peritonealcavity macrophages [88]. This is likely to be the right way forward, since many experimental protocols devised to study the in vivo stimulatory potential and the molecular mechanisms involved in antigen processing of DC require large numbers of DC, approximately (10 8 -10 9 ).…”

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

“…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].…”

“…PP comprise transmucosal clusters of lymphoid follicles in the small intestine. In rats, PP are usually 3-4 mm long [14,18,26] but in large animals, for example cattle, PP may extend up to 4 m along the intestine [9]. Studies in pig, sheep, cattle have shown that there are two separate types of PP, which are morphologically different: discrete Peyer's Patches (DPP) scattered throughout the jejunum and a large continuous patch in the terminal ileum [9,20,27,38].…”

“…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.…”