Sárváry Attila, Szucs Sándor, Balogh Imre, Becsky Aron, Bárdos Helga, Kávai Mária, Seligsohn Uri, Egbring Rudolf, Lopaciuk Stanislaw, Muszbek László, Adány Róza
Department of Preventive Medicine, School of Public Health, Faculty of Medicine, Medical and Health Science Center, University of Debrecen, Debrecen H-4012, P.O. Box 9, Hungary.
Cell Immunol. 2004 Apr;228(2):81-90. doi: 10.1016/j.cellimm.2004.04.007.
Besides its traditional role in hemostasis, factor XIII subunit A (FXIII-A) is supposed to function as a cellular transglutaminase and to be involved in certain intracellular processes, including cytoskeletal remodeling. To investigate its intracellular role, the aim of the present study was to follow changes in FXIII-A production in combination with the receptor-mediated phagocytic activities of monocytes/macrophages and to examine the phagocytic functions of monocytes in patients with FXIII-A deficiency. Human blood monocytes were isolated from the buffy coats of healthy volunteers and cultured for 4 days. The FcgammaR-mediated phagocytosis of sensitized erythrocytes (EA) and the complement receptor (CR)-mediated phagocytosis of complement-coated yeast particles were studied during monocyte/macrophage differentiation. Changes in the gene expression of FXIII-A were detected by real-time quantitative RT-PCR. FXIII-A protein production was investigated with fluorescent image analysis at single cell level and Western immunoblot analysis. Both the FcgammaR and CR-mediated phagocytosis increased during culturing, which peaked on day 3. The phagocytic activity of the cells could be markedly inhibited with monodansylcadaverine, an inhibitor of the transglutaminase-induced crosslinking of proteins. The phagocytosis of EA, complement-coated and uncoated yeast particles was found to be strongly diminished in monocytes of FXIII-A deficient patients. The phagocytic functions of cultured cells showed a change in parallel with the alterations in FXIII-A mRNA expression, as well as with that in FXIII-A in protein synthesis detected by image and Western immunoblot analyses in concert. Our results suggest that FXIII-A plays a role in the Fcgamma and complement receptor-mediated phagocytic activities of monocytes/macrophages.
除了在止血方面的传统作用外，凝血因子 XIII A 亚基（FXIII-A）被认为具有细胞转谷氨酰胺酶的功能，并参与某些细胞内过程，包括细胞骨架重塑。为了研究其在细胞内的作用，本研究的目的是追踪 FXIII-A 产生的变化，并结合单核细胞/巨噬细胞的受体介导吞噬活性，同时检查 FXIII-A 缺乏患者单核细胞的吞噬功能。从健康志愿者的血沉棕黄层中分离出人类血液单核细胞，并培养 4 天。在单核细胞/巨噬细胞分化过程中，研究了 FcγR 介导的致敏红细胞（EA）吞噬作用以及补体受体（CR）介导的补体包被酵母颗粒吞噬作用。通过实时定量 RT-PCR 检测 FXIII-A 的基因表达变化。在单细胞水平上用荧光图像分析和 Western 免疫印迹分析研究 FXIII-A 蛋白的产生。在培养过程中，FcγR 和 CR 介导的吞噬作用均增加，在第 3 天达到峰值。转谷氨酰胺酶诱导的蛋白质交联抑制剂单丹磺酰尸胺可显著抑制细胞的吞噬活性。发现 FXIII-A 缺乏患者的单核细胞中，EA、补体包被和未包被酵母颗粒的吞噬作用明显减弱。培养细胞的吞噬功能变化与 FXIII-A mRNA 表达的改变以及图像分析和 Western 免疫印迹分析检测到的 FXIII-A 蛋白合成的改变平行。我们的结果表明，FXIII-A 在单核细胞/巨噬细胞的 Fcγ 和补体受体介导的吞噬活性中起作用。
