Lipsky P E, Ziff M
J Clin Invest. 1980 May;65(5):1069-76. doi: 10.1172/JCI109759.
The effect of d-penicillamine (Pen) and mixtures of Pen and copper sulfate on the capacity of normal human peripheral blood mononuclear cells (PBM) to generate immunoglobulin-secreting cells (ISC) in response to the T-cell-dependent polyclonal B-cell activators pokeweed mitogen (PWM) and staphylococcal protein A (SPA) was examined. PBM obtained from normal individuals were incubated for 1-2 h at 37 degrees C with medium alone, Pen, CuSO(4), or a mixture of Pen and CuSO(4). After washing, the cells were incubated for 6-7 d with PWM or SPA and then, with a reverse hemolytic plaque assay, assayed for the number of ISC generated. Preincubation of PBM with either Pen (100 mug/ml) or CuSO(4) (2 mug/ml) did not alter the subsequent capacity of the cells to generate ISC in response to PWM or SPA. In contrast, responsiveness to both mitogens was nearly abolished when PBM were similarly preincubated with a mixture of Pen and CuSO(4). Inhibition of responsiveness could not be ascribed to cell death, carry-over of the inhibitors, or an alteration in the concentration of PWM or the length of incubation yielding maximum responses. Co-culture experiments demonstrated that Pen and CuSO(4) preincubation had not caused augmented suppressor cell function. Experiments in which PBM were separated into adherent and nonadherent populations indicated that Pen and CuSO(4) preincubation inhibited the responsiveness of the nonadherent cells but did not alter the accessory cell function of monocytes. To determine whether Pen and CuSO(4) preincubation effected T- or B-cell function, PBM were separated into B- and T-cell-enriched populations, individually preincubated with Pen and CuSO(4), and then co-cultured with PWM. The results indicated that Pen and CuSO(4) markedly inhibited helper T-cell function and had little effect on the capacity of B cells to generate ISC. The observation that in the presence of CuSO(4) Pen inhibits helper T-cell activity may, in part, explain the therapeutic efficacy of Pen in rheumatoid arthritis and especially the capacity of Pen therapy to decrease antiglobulin titers in treated patients.
研究了d-青霉胺（Pen）以及Pen与硫酸铜的混合物对正常人外周血单个核细胞（PBM）在T细胞依赖性多克隆B细胞激活剂商陆有丝分裂原（PWM）和葡萄球菌蛋白A（SPA）刺激下产生免疫球蛋白分泌细胞（ISC）能力的影响。从正常个体获取的PBM在37℃下分别与单独的培养基、Pen、CuSO₄或Pen与CuSO₄的混合物孵育1 - 2小时。洗涤后，细胞与PWM或SPA孵育6 - 7天，然后通过反向溶血空斑试验检测产生的ISC数量。PBM预先用Pen（100μg/ml）或CuSO₄（2μg/ml）孵育，并不改变细胞随后对PWM或SPA产生ISC的能力。相反，当PBM用Pen和CuSO₄的混合物进行类似的预先孵育时，对两种有丝分裂原的反应性几乎完全丧失。反应性的抑制不能归因于细胞死亡、抑制剂的残留、PWM浓度的改变或产生最大反应的孵育时间的改变。共培养实验表明，Pen和CuSO₄预先孵育并未导致抑制性细胞功能增强。将PBM分为贴壁细胞群和非贴壁细胞群的实验表明，Pen和CuSO₄预先孵育抑制了非贴壁细胞的反应性，但未改变单核细胞的辅助细胞功能。为了确定Pen和CuSO₄预先孵育是否影响T细胞或B细胞功能，将PBM分为富含B细胞和T细胞的群体，分别用Pen和CuSO₄预先孵育，然后与PWM共培养。结果表明，Pen和CuSO₄显著抑制辅助性T细胞功能，而对B细胞产生ISC的能力影响很小。在CuSO₄存在下Pen抑制辅助性T细胞活性这一观察结果，可能部分解释了Pen在类风湿性关节炎中的治疗效果，尤其是Pen治疗降低治疗患者抗球蛋白滴度 的能力。
