Montgomery R R, Nathanson M H, Malawista S E
Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06510.
J Immunol. 1993 Feb 1;150(3):909-15.
The macrophage is a known reservoir for a number of infectious agents, and is therefore a likely candidate site for persistence of Borrelia burgdorferi, the Lyme spirochete. We report that unopsonized B. burgdorferi enter macrophages rapidly, resulting mainly in degradation but occasionally in apparent intracellular persistence. We studied uptake of spirochetes by macrophages by simultaneously labeling infected cells with antibodies to B. burgdorferi and with sequential components of the endocytic pathway, and we examined optical sections (0.5-1.0 micron in thickness) of these cells by confocal fluorescence microscopy at multiple time points after infection. We found that only 5 min of incubation at 37 degrees C were required for nearly 100% of B. burgdorferi to enter a lysosomal glycoprotein-positive compartment, whereas 60 min were required for 90% of the spirochetes to appear in a cathepsin L-positive compartment under the same conditions. We also labeled infected living cells with acridine orange to distinguish live from killed intracellular organisms. Although the large majority of spirochetes within a given cell were dead, we saw occasional live ones up to 24 h (the longest interval examined) after all extracellular organisms had been lysed in distilled water. Moreover, we can reculture spirochetes from macrophages after infection. Persistence of spirochetes within macrophages provides a possible pathogenetic mechanism for chronic or recurrent Lyme disease in man.
巨噬细胞是多种感染因子的已知储存库,因此是莱姆螺旋体——伯氏疏螺旋体持续存在的可能候选位点。我们报告称,未被调理素包被的伯氏疏螺旋体可迅速进入巨噬细胞,主要导致其降解,但偶尔也会出现明显的细胞内持续存在现象。我们通过用抗伯氏疏螺旋体抗体和内吞途径的连续成分同时标记感染细胞,研究了巨噬细胞对螺旋体的摄取,并在感染后的多个时间点通过共聚焦荧光显微镜检查了这些细胞的光学切片(厚度为0.5 - 1.0微米)。我们发现,在37℃孵育仅5分钟,近100%的伯氏疏螺旋体就会进入溶酶体糖蛋白阳性区室,而在相同条件下,90%的螺旋体需要60分钟才会出现在组织蛋白酶L阳性区室。我们还用吖啶橙标记感染的活细胞,以区分活的和死亡的细胞内生物体。尽管给定细胞内的绝大多数螺旋体已死亡,但在所有细胞外生物体在蒸馏水中被裂解后,我们偶尔能看到长达24小时(所检测的最长时间间隔)的活螺旋体。此外,感染后我们可以从巨噬细胞中重新培养出螺旋体来。螺旋体在巨噬细胞内的持续存在为人类慢性或复发性莱姆病提供了一种可能的致病机制。