Jandl R C, George J L, Silberstein D S, Eaton R B, Schur P H
Clin Immunol Immunopathol. 1987 Mar;42(3):344-59. doi: 10.1016/0090-1229(87)90023-7.
The role of adherent cells in the regulation of anti-DNA and immunoglobulin synthesis was investigated in patients with systemic lupus erythematosus (SLE). Peripheral blood mononuclear cells were cultured for 7 days, or were washed after 20 hr of incubation and recultured in fresh media. This washing resulted in a marked decrease in total IgG, IgM, and anti-DNA synthesis compared with unwashed cultures. Reculturing the washed cells in their original supernatant reconstituted Ig and anti-DNA synthesis. Hence it appeared that a supernatant factor, or factors, present in the first 20 hr of mononuclear cell cultures, was required for maximal Ig synthesis. Adherent cells were found to be the source of this Ig-stimulating activity. Moreover, adherent cell supernatants had no direct Ig-stimulating effect on B cells. Ig synthesis was stimulated, however, when T cells where present with the B cells at a 3:1 ratio. Autologous SLE mononuclear cell supernatants reconstituted Ig synthesis to a greater degree than did autologous normal supernatants. SLE adherent cell supernatants were fractionated on an HPLC sizing column. The fractions were tested for their ability to stimulate IgG synthesis by SLE mononuclear cells that had been washed after 20 hr of culture. A single peak of IgG-stimulating activity was found at approximately 14,000 Mr. A rabbit antiserum to interleukin-1 (IL-1) neutralized the Ig-stimulating activity in adherent cell supernatants. No correlations were found, however, between supernatant IL-1 levels assayed by C3H-HeJ mouse thymocyte proliferation and IgG stimulation in mononuclear cell cultures, suggesting that the effects of IL-1 on cell proliferation may not accurately reflect its effects on Ig synthesis. These observations suggest that in normal individuals and in patients with SLE in vitro polyclonal Ig and anti-DNA synthesis requires the presence of soluble adherent cell factors. The Ig-stimulating effect is facilitated by T cells and appears to be mediated at least in part by IL-1. This culture technique provides a new way of analyzing the role of soluble factors in autoantibody synthesis and suggests that IL-1 may be an important contributor to lupus B-cell hyperactivity.
在系统性红斑狼疮（SLE）患者中，研究了贴壁细胞在抗DNA和免疫球蛋白合成调节中的作用。外周血单个核细胞培养7天，或在培养20小时后洗涤，然后在新鲜培养基中重新培养。与未洗涤的培养物相比，这种洗涤导致总IgG、IgM和抗DNA合成显著减少。将洗涤后的细胞在其原始上清液中重新培养可恢复Ig和抗DNA合成。因此，似乎在单核细胞培养的最初20小时内存在的一种或多种上清液因子是最大Ig合成所必需的。发现贴壁细胞是这种Ig刺激活性的来源。此外，贴壁细胞上清液对B细胞没有直接的Ig刺激作用。然而，当T细胞与B细胞以3:1的比例存在时，Ig合成受到刺激。自体SLE单个核细胞上清液比自体正常上清液在更大程度上恢复了Ig合成。SLE贴壁细胞上清液在HPLC尺寸排阻柱上进行分级分离。测试各组分刺激培养20小时后洗涤过的SLE单个核细胞合成IgG的能力。在约14,000 Mr处发现了一个IgG刺激活性的单峰。抗白细胞介素-1（IL-1）的兔抗血清中和了贴壁细胞上清液中的Ig刺激活性。然而，通过C3H-HeJ小鼠胸腺细胞增殖测定的上清液IL-1水平与单核细胞培养中的IgG刺激之间未发现相关性，这表明IL-1对细胞增殖的作用可能无法准确反映其对Ig合成的作用。这些观察结果表明，在正常个体和SLE患者中，体外多克隆Ig和抗DNA合成需要可溶性贴壁细胞因子的存在。T细胞促进了Ig刺激作用，并且似乎至少部分由IL-1介导。这种培养技术为分析可溶性因子在自身抗体合成中的作用提供了一种新方法，并表明IL-1可能是狼疮B细胞过度活跃的重要促成因素。
