Kennedy M S, Stobo J D, Goldyne M E
Prostaglandins. 1980 Jul;20(1):135-45. doi: 10.1016/0090-6980(80)90013-1.
This report focuses on the identification of the human peripheral blood mononuclear cells that do or do not produce prostaglandins (PGs) and related arachidonic acid metabolites. Our results, using two different assay systems, indicate that the monocyte/macrophage (Mphi) is the major and possibly sole source of thromboxane (TXB2) and prostaglandin E2 (PGE2) among peripheral blood mononuclear cells. Adherent peripheral blood monocytes (> 95% esterase positive) produced substantial amounts of these compounds. Quantitation of products which had incorporated exogenous 14C-arachidonic acid and radioimmunoassay of adherent cell culture fluids demonstrated that the amount of TXB2 produced by these cells was appreciably greater than the amount of PGE2 produced. Additional confirmation of TXB2 synthesis was shown by abolishing the TXB2 peak on TLC and TXB2 activity detected by RIA by treating cells with a specific inhibitor of thromboxane synthetase. In contrast, non-adherent T cells failed to synthesize either PGE2 or TXB2. Non-adherent B cells (95% Ig positive) incubated with 14C-arachidonic acid produced a small peak of radioactivity co-chromatographing with TXB2, and no PGE. All three cell populations incorporated similar amounts of 14C-arachidonic acid into hydroxy-fatty acids. We were unable to detect 6-keto-F1 alpha, the hydrolysis product of prostacyclin (PGI2) in any of the cell types tested. The absence of PG synthesis among normal peripheral blood T and B cells was also noted among established human lymphoid cell lines. Neither a human T (CCRF), nor a human B-cell line (GM-130), produced PGE2 or TXB2. Three murine macrophage cell lines, P388D1, J774.2, and WEHI-3 produced PGE2 and the latter TXB2 as well.
本报告重点关注产生或不产生前列腺素（PGs）及相关花生四烯酸代谢产物的人外周血单个核细胞的鉴定。我们使用两种不同检测系统得出的结果表明，在外周血单个核细胞中，单核细胞/巨噬细胞（Mphi）是血栓素（TXB2）和前列腺素E2（PGE2）的主要且可能是唯一来源。贴壁外周血单核细胞（酯酶阳性率>95%）产生大量此类化合物。对掺入外源性14C - 花生四烯酸的产物进行定量以及对贴壁细胞培养液进行放射免疫测定表明，这些细胞产生的TXB2量明显大于PGE2量。通过用血栓素合成酶特异性抑制剂处理细胞，使薄层层析（TLC）上的TXB2峰消失以及放射免疫分析（RIA）检测到的TXB2活性消失，进一步证实了TXB2的合成。相比之下，非贴壁T细胞无法合成PGE2或TXB2。用14C - 花生四烯酸孵育的非贴壁B细胞（Ig阳性率95%）产生了一个与TXB2共色谱的小放射性峰，且未产生PGE。所有三种细胞群体将相似量的14C - 花生四烯酸掺入羟基脂肪酸中。在任何测试的细胞类型中，我们均未检测到前列环素（PGI2）的水解产物6 - 酮 - F1α。在已建立的人淋巴细胞系中也观察到正常外周血T细胞和B细胞中缺乏PG合成。无论是人T细胞系（CCRF）还是人B细胞系（GM - 130），均不产生PGE2或TXB2。三种鼠巨噬细胞系P388D1、J774.2和WEHI - 3产生PGE2，后者还产生TXB2。
