Ohosone Y, Akizuki M, Hirakata M, Satoh M, Yamagata H, Homma M
Arthritis Rheum. 1986 Nov;29(11):1343-50. doi: 10.1002/art.1780291106.
Antibodies to U1 ribonucleoproteins produced by peripheral blood mononuclear cells from patients with connective tissue diseases were measured by a sensitive enzyme-linked immunosorbent assay and by immunoblotting. Mononuclear cells from patients with high-titer serum antibodies spontaneously started secreting IgG anti-U1 RNP antibodies on the second day after culture. The amount of anti-U1 RNP in the culture supernatants reached maximum level on day 8. The most effective production of anti-U1 RNP by B cells was observed when they were cultured in the presence of T cells and adherent cells. Mononuclear cells from patients without anti-U1 RNP antibodies or from normal subjects did not produce a measurable amount of anti-U1 RNP. Treatment of mononuclear cells by cycloheximide resulted in complete inhibition of anti-U1 RNP secretion, which indicates that antibody in the culture supernatants reflects the active biologic phenomenon in vitro. The methods described should be useful in the study of the cellular mechanisms involved in antinuclear antibody production of connective tissue diseases.
通过灵敏的酶联免疫吸附测定法和免疫印迹法，检测结缔组织病患者外周血单个核细胞产生的抗U1核糖核蛋白抗体。来自血清抗体滴度高的患者的单个核细胞在培养后的第二天开始自发分泌IgG抗U1 RNP抗体。培养上清液中抗U1 RNP的量在第8天达到最高水平。当B细胞在T细胞和贴壁细胞存在的情况下培养时，观察到B细胞产生抗U1 RNP的效率最高。来自无抗U1 RNP抗体患者或正常受试者的单个核细胞不产生可测量量的抗U1 RNP。用环己酰亚胺处理单个核细胞会导致抗U1 RNP分泌完全受到抑制，这表明培养上清液中的抗体反映了体外活跃的生物学现象。所描述的方法应有助于研究结缔组织病抗核抗体产生所涉及的细胞机制。
