Klaesson S, Tammik L, Markling L, Lundkvist I, Ringdén O
Department of Transplantation Surgery, Karolinska Institute, Huddinge Hospital, Stockholm, Sweden.
Scand J Immunol. 1996 May;43(5):574-82. doi: 10.1046/j.1365-3083.1996.d01-72.x.
Antibody-secreting B cells were measured as plaque-forming cells (PFC) in a modified haemolysis-in-gel assay, using protein A coupled sheep erythrocytes as targets. Human lymphocytes from blood (PBL), bone marrow or spleen were stimulated in vitro by various polyclonal B-cell activators and incubated with intravenous immunoglobulin (IVIG) or peptide fragments of IVIG. IgG and IgM production from PBL and bone marrow cells, measured as PFC, was inhibited more than 50% by IVIG 2.5 mg/ml, compared to controls without IVIG. Inhibition of the IgG and IgM response of spleen cells by IVIG varied depending on the stimuli. Using Staphylococcus aureus protein A (SPA), inhibition was almost 90% (P < 0.001). The inhibition of the IgG and IgM responses to lipopolysaccharide from Escherichia coli (LPS) were 70% (P < 0.01) and 28% (P < 0.05), respectively. IgG stimulation by pokeweed mitogen (PWM) was inhibited by 57% (P < 0.01), but the IgM response was inhibited only by the higher IVIG concentration of 5.0 mg/ml. In mixed lymphocyte cultures of spleen cells, IgG and IgM production were inhibited by more than 60% (P < 0.05). The effect of IgG, IgG-F(ab')2 and IgG-Fc on LPS or PWM-stimulated spleen cells were compared, using equimolar concentrations of the various preparations. IgG- and IgM-producing PFC were significantly (P < 0.05) inhibited in a dose-dependent fashion by IgG and F(ab')2, but not by Fc. LPS-induced IgG and IgM production was inhibited also when IgG and F(ab')2 were added up to 48 h after the stimulator. A comparison of IgG, F(ab')2 and Fc products from different companies showed that all IgG and F(ab')2 preparations significantly inhibited IgG and IgM production of LPS-stimulated spleen cells. No significant inhibition was obtained with any of the purified Fc products.
在改良的凝胶内溶血试验中，以形成噬斑细胞（PFC）来测定分泌抗体的B细胞，使用与蛋白A偶联的绵羊红细胞作为靶标。来自血液（外周血淋巴细胞，PBL）、骨髓或脾脏的人淋巴细胞，通过各种多克隆B细胞激活剂在体外进行刺激，并与静脉注射免疫球蛋白（IVIG）或IVIG的肽片段一起孵育。与未添加IVIG的对照组相比，2.5 mg/ml的IVIG可使PBL和骨髓细胞产生的IgG和IgM（以PFC衡量）的生成受到超过50%的抑制。IVIG对脾脏细胞IgG和IgM反应的抑制作用因刺激物而异。使用金黄色葡萄球菌蛋白A（SPA）时，抑制率近90%（P<0.001）。对来自大肠杆菌的脂多糖（LPS）的IgG和IgM反应的抑制率分别为70%（P<0.01）和28%（P<0.05）。美洲商陆丝裂原（PWM）刺激的IgG生成受到57%的抑制（P<0.01），但只有在IVIG浓度较高的5.0 mg/ml时，IgM反应才受到抑制。在脾脏细胞的混合淋巴细胞培养中，IgG和IgM的生成受到超过60%的抑制（P<0.05）。使用等摩尔浓度的各种制剂，比较了IgG、IgG-F(ab')2和IgG-Fc对LPS或PWM刺激的脾脏细胞的影响。IgG和F(ab')2以剂量依赖性方式显著（P<0.05）抑制产生IgG和IgM的PFC，但Fc则无此作用。当在刺激物加入后长达48小时添加IgG和F(ab')2时，LPS诱导的IgG和IgM生成也受到抑制。对不同公司的IgG、F(ab')2和Fc产品进行比较表明，所有IgG和F(ab')2制剂均显著抑制LPS刺激的脾脏细胞的IgG和IgM生成。任何纯化的Fc产品均未获得显著抑制作用。
