Lyme disease spirochetes induce human and murine interleukin 1 production.
Lyme disease spirochetes induce human and murine interleukin 1 production.
复制标题
莱姆病螺旋体诱导人和鼠白细胞介素 1 的产生。
作者：
G. Habicht;G. Beck;J. Benach;J. Coleman;K. Leichtling
IL 1是巨噬细胞产生的一种主要的免疫调节分子，它似乎是非特异性宿主防御机制对各种环境损伤的分子协调者。许多研究者已经使用人工试剂刺激巨噬细胞产生IL-1。我们现在报道在生理刺激后产生大量的IL-1。莱姆病螺旋体，最近分离和适应体外生长，用于刺激P388 D1细胞或人外周血单核细胞。将螺旋体以10：1的比例加入到无血清RPMI中的融合巨噬细胞培养物中。IL 1的释放呈剂量依赖性。用胸腺细胞ConA共分裂试验测定24小时培养上清IL 1活性。活性不是由于螺旋体上的内毒素或由螺旋体产生的。多粘菌素B亲和柱不能去除活性，螺旋体-巨噬细胞培养物中的多粘菌素B对IL 1的产生没有影响。收集上清液，浓缩，并进行尺寸排阻HPLC。在P388 D1细胞上清液中发现三个活性区域（Mr大于60，000、40，000和20，000），而在人单核细胞上清液中发现两个峰（Mr 23，000和13，000）。分别对来自鼠和人细胞上清液的Mr 20，000和13，000峰进行SDS-PAGE，显示为单一条带（小鼠IL 1的Mr 12，400和人IL 1的Mr 13，500）。柱纯化的IL 1制备物的等电聚焦显示人IL 1（pI 7.25和4.4至5）和鼠IL 1（pI 7.25和5.55）的两种不同pI。与鼠或人IL 1制剂一起培养的成纤维细胞表现出分泌的胶原酶增加和细胞增殖增加。因此，生理刺激和简单的生物化学技术产生大量非常纯的小鼠或人IL 1。这种IL 1是由莱姆病螺旋体刺激的巨噬细胞产生的，这可能解释了莱姆病的一些临床表现。
IL 1 is a major immunoregulatory molecule produced by macrophages, and it appears to be the molecular orchestrator of nonspecific host defense mechanisms against a variety of environmental insults. Many investigators have used artificial agents to stimulate macrophages to produce IL 1. We now report production of large quantities of IL 1 after a physiologic stimulus. The Lyme disease spirochete, recently isolated and adapted for growth in vitro, was used to stimulate P388D1 cells or human peripheral blood monocytes. Spirochetes were added to confluent macrophage cultures in serum-free RPMI at a ratio of 10:1. The release of IL 1 was dose-dependent. The 24-hr supernatant IL 1 activity was determined by using the thymocyte Con A co-mitogenesis assay. Activity was not due to an endotoxin on, or produced by, the spirochete. A polymyxin B affinity column failed to remove activity, and polymyxin B in the spirochete-macrophage culture had no effect on IL 1 production. Supernatants were collected, were concentrated, and were subjected to size exclusion HPLC. Three areas of activity were found in P388D1 cell supernatants (Mr greater than 60,000, 40,000, and 20,000), whereas two peaks (Mr 23,000 and 13,000) were found in human monocyte supernatants. The Mr 20,000 and 13,000 peaks from murine and human cell supernatants, respectively, were subjected to SDS-PAGE and were shown to be single bands (Mr 12,400 for the mouse IL 1 and Mr 13,500 for the human IL 1). Isoelectric focusing of column-purified IL 1 preparations showed two different pI in both human (pI 7.25 and 4.4 to 5) and murine (pI 7.25 and 5.55) IL 1. Fibroblasts cultured with murine or human IL 1 preparations demonstrated both an increase in secreted collagenase and increased cell proliferation. Thus, a physiologic stimulus and simple biochemical techniques produce large amounts of very pure mouse or human IL 1. That this IL 1 is produced by Lyme disease spirochete-stimulated macrophages may explain some of the clinical manifestations of Lyme disease.