Isobe K, Nakashima I
Department of Immunology, Nagoya University School of Medicine, Japan.
J Cell Biochem. 1993 Nov;53(3):198-205. doi: 10.1002/jcb.240530304.
The indirect stimulation of macrophages to produce nitrite was examined by using the macrophage cell line J774.J774 spontaneously produced nitrite, when cultured at high concentration. J774 cultured in low concentration (< 10(4) cells in 100 microliters) barely produced nitrite. J774 cultured in low concentration produced a large amount of nitrite by the co-culture of nonadherent spleen cells or nonadherent peritoneal exudate cells, which were stimulated with con A, anti-CD3, or staphylococcal enterotoxin A. J774 (BALB/c derived: H-2d) cultured with either syngeneic (BALB/c) or allogeneic (B6; H-2b B10BR; H-2k) nonadherent lymphocytes, which were stimulated with conA or anti-CD3, produced nitric oxide. However, J774 produced nitric oxide by stimulation with SEA only when co-cultured with SEA-reactive T lymphocytes. Peritoneal exudate cells from mice, which did not proliferate by the stimulation of conA or anti-CD3, proliferated well by the addition of L-arginine homologue, NG-monomethyl-L-arginine. The proliferation of nonadherent peritoneal exudate cells stimulated with conA or anti-CD3 was suppressed by the addition of peritoneal macrophages. This suppression was abolished by the addition of NG-monomethyl-L-arginine.
利用巨噬细胞系J774研究了间接刺激巨噬细胞产生亚硝酸盐的情况。J774在高浓度培养时会自发产生亚硝酸盐。在低浓度（100微升中<10⁴个细胞）培养的J774几乎不产生亚硝酸盐。在低浓度培养的J774通过与经刀豆蛋白A（Con A）、抗CD3或葡萄球菌肠毒素A刺激的非贴壁脾细胞或非贴壁腹腔渗出细胞共培养，会产生大量亚硝酸盐。与经Con A或抗CD3刺激的同基因（BALB/c）或异基因（B6；H-2b B10BR；H-2k）非贴壁淋巴细胞共培养的J774（源自BALB/c：H-2d）会产生一氧化氮。然而，J774仅在与SEA反应性T淋巴细胞共培养时通过SEA刺激产生一氧化氮。经Con A或抗CD3刺激不增殖的小鼠腹腔渗出细胞，通过添加L-精氨酸同系物NG-单甲基-L-精氨酸后增殖良好。添加腹腔巨噬细胞会抑制经Con A或抗CD3刺激的非贴壁腹腔渗出细胞的增殖。添加NG-单甲基-L-精氨酸可消除这种抑制作用。
