Shapira L, Soskolne W A, Van Dyke T E
Department of Periodontology, Hebrew University-Hadassah Faculty of Dental Medicine, Jerusalem, Israel.
J Periodontol. 1996 Mar;67(3):224-8. doi: 10.1902/jop.1996.67.3.224.
Previous studies from our laboratory have demonstrated that freshly isolated peripheral blood monocytes from localized juvenile periodontitis (LJP) patients secrete more prostaglandin E2 (PGE2) after stimulation by lipopolysaccharide (LPS) than do monocytes from healthy subjects. However, it is not clear if the altered function of LJP monocytes is intrinsic to the cells or is induced by the persistent infection of the periodontium. The present study was designed to compare PGE2 secretion by freshly-isolated peripheral blood monocytes (FIM) from LJP and control subjects to in vitro monocyte-derived macrophages (MDM) from the same subjects. We also examined monocyte maturation into macrophage-like cells and the cell-surface expression of the LPS, receptor, CD14 during the culture period. FIM from LJP patients and controls were stimulated by different concentrations of LPS (O to 30 micrograms/ml) for 24 hours. These experiments were performed immediately after cell separation and after 10 days in culture, which allowed differentiation of monocytes into MDM. PGE2 levels in the culture media were determined using a radioimmunoassay. Cell surface expression of CD71, a cell maturation marker, and CD14 were assayed by cell-ELISA in relation to beta-2-microglobulin. LPS-stimulated FIM from LJP patients secreted 3 to 4 times more PGE2 than control FIM at all LPS concentrations tested. After 10 days in culture, the LJP MDM secretion of PGE2 reduced to control MDM level of PGE2 secretion. These levels of PGE2 secretion were comparable to PGE2 secretion from FIM of controls. Cell maturation, as verified by CD71 expression, was found not to differ between the groups. However, the expression of CD14 by LJP FIM was much lower than on control FIM (approximately equal to 50%). After 5 or 10 days in culture, MDM from both control and LJP subjects expressed comparable amounts of CD14. The results suggest that in vitro conditions reverse the hypersensitivity of LJP monocytes to LPS into control levels and CD14 expression is not correlated to the hyper-responsiveness of the cells to LPS.
我们实验室之前的研究表明,与健康受试者的单核细胞相比,从局限性青少年牙周炎(LJP)患者新鲜分离的外周血单核细胞在受到脂多糖(LPS)刺激后分泌更多的前列腺素E2(PGE2)。然而,尚不清楚LJP单核细胞功能的改变是细胞固有的,还是由牙周组织的持续感染所诱导。本研究旨在比较LJP患者和对照受试者新鲜分离的外周血单核细胞(FIM)与来自相同受试者的体外单核细胞衍生巨噬细胞(MDM)的PGE2分泌情况。我们还检测了培养期间单核细胞向巨噬细胞样细胞的成熟情况以及LPS受体CD14的细胞表面表达。LJP患者和对照的FIM用不同浓度的LPS(0至30微克/毫升)刺激24小时。这些实验在细胞分离后立即进行,并在培养10天后进行,这使得单核细胞分化为MDM。使用放射免疫测定法测定培养基中的PGE2水平。通过细胞ELISA检测细胞成熟标志物CD71和CD14相对于β2-微球蛋白的细胞表面表达。在所有测试的LPS浓度下,LJP患者的LPS刺激FIM分泌的PGE2比对照FIM多3至4倍。培养10天后,LJP MDM的PGE2分泌降至对照MDM的PGE2分泌水平。这些PGE2分泌水平与对照FIM的PGE2分泌相当。通过CD71表达验证,发现两组之间的细胞成熟情况没有差异。然而,LJP FIM的CD14表达远低于对照FIM(约为50%)。培养5天或10天后,对照和LJP受试者的MDM表达的CD14量相当。结果表明,体外条件可将LJP单核细胞对LPS的超敏反应逆转至对照水平,且CD14表达与细胞对LPS的高反应性无关。