AN EXPERIMENTAL STUDY OF THE HISTOGENESIS OF THE MILIARY TUBERCLE IN VITALLY STAINED RABBITS.
AN EXPERIMENTAL STUDY OF THE HISTOGENESIS OF THE MILIARY TUBERCLE IN VITALLY STAINED RABBITS.
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DOI：
10.1084/jem.19.3.283
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发表时间：
1914-03-01
期刊：
影响因子：
--
通讯作者：
Winternitz MC
中科院分区：
文献类型：
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作者：
Evans HM;Bowman FB;Winternitz MC
在我们对接种牛结核病时被活体染色的动物肝小叶内发育的粟粒结节的组织发生进行的研究中，对照使我们能够认识到活体染色影响肝脏的方式。因此，不可能将生物体的影响与染料的影响混淆。然而，有趣的是，这些影响是密切相关的。仅活体染色就能逐渐产生某些变化，这些变化在实验性结核病中以快得多的速度发生。虽然结核动物的枯否细胞在几小时内开始对疾病产生反应，但正常动物的枯否细胞在连续每日注射第三或第四剂后才出现活性染色。然而，在许多天后，仅活体染色就引起枯否细胞的增大、增殖和分离，使得这些枯否细胞转化为可能具有一个或多个核的大的游离吞噬细胞。这些是慢性染色动物的巨大巨噬细胞。在我们所有的实验中，我们只使用了急性染色的动物，因此染料本身的影响永远不足以产生变化。事实上，没有证据表明染料在实验期间明显加重了这些变化。然而，染料却能向我们显示进入结核性肉芽肿的细胞类型，因为当大量的台盼蓝进入体液时，它就能进入所有能接受它的细胞，活体染色可以说是对细胞的一种生理测试。无论台盼蓝和联苯胺染料产生的活体染色的基本性质如何，重要的是，这种反应不会在任何可感知的程度上发生在单核血细胞上，而且它确实会在肝内皮细胞上明显发生。通过这种活体试验，当结核菌悬液进入门静脉血流时，会出现以下现象。这些生物被血流扫过，最后停留在门静脉的末端分支，在那里它们堵塞血管并继续繁殖。它们损伤血管壁并在其周围引起渗出性炎症过程，最终导致不仅位于这些区域而且位于肝小叶内的结节的形成。血管壁的损伤在早期阶段通过其结构中存在有生命力的染色区域来表现。在半小时结束时，发现细菌在较大血管中成团地存在于细胞外，但在一定程度上已经存在于整个肝脏的染色的枯否细胞体内。渗出性炎症表现为多核白细胞在细菌团块周围的短暂聚集，这可能早在接种后半小时就可以看到。它们继续存在于门静脉周围区域的较大细胞团中许多天，但它们很快被其他单核细胞取代，因此在一天内门静脉栓的组织学外观发生了根本性变化。因此，最活跃地进入反应的单核细胞是内皮细胞，而不是血原细胞，活体染色使我们能够明确区分。这一事实在门静脉栓塞中是明显的，在肝小叶内结节形成的病例中也是明确的。这种结节很可能是由库普弗细胞内的单个生物体的定位引起的，因为我们已经发现了这种可能的循环的初始阶段。它们包括在某些枯否细胞中发生有丝分裂，其中Ziehl-Nielson方法显示一个或几个杆菌被吞噬（图4）。此时，受感染的细胞迅速生长，36小时后，产生的多核巨细胞大部分与血管壁的其他内皮细胞分离。许多杆菌存在于这些细胞的原生质中（图5），它们对活体染色的强烈反应特别明显。他们接受了过量的台盼蓝，以至于在36小时阶段的肝脏切片的低倍视图显示这些细胞为深蓝色斑点（图8）。巨细胞起源于库普弗细胞，这一点不仅从上述序列和选择性染色中可以明显看出，而且从这样一个事实中也可以明显看出，即使在完全形成时，原生质束仍然将其连接到其母组织--正常的血管内皮。这些丝缠绕在其他细胞的网眼中，特别是单核血细胞，其中一种是多胚型的，具有同质的原生质，在有丝分裂中并不少见。这些细胞在结核病的损害中无疑是重要的。我们在接种后不久就在毛细血管中大量看到它们，它们也单独出现或在枯否细胞和肝柱之间的巢中出现。一般来说，它们不含生命染料。他们继续关注结节的进一步生长，并与结缔组织细胞一起使老年结节的结构相对复杂。另一方面，年轻的小叶内肿块的结构几乎不复杂。36小时后形成的粟粒性结节由一个巨细胞组成，周围是上皮样细胞和上述多胚型血细胞。巨细胞及其所谓的上皮样细胞被选择性染色，并且仅来源于肝内皮。
In our study of the histogenesis of the miliary tubercle developing inside the liver lobule in animals that have been stained vitally while inoculated with bovine tuberculosis, the controls enable us to recognize the manner in which the vital stain affects the liver. There is therefore no possibility of confusing the effects due to the organism with the effects due to the dye. It is, however, of interest to note that the effects are closely related. The vital stain alone is able to produce gradually some of the same changes that occur with far greater rapidity in experimental tuberculosis. Although in a few hours the Kupffer cells of tuberculous animals begin to react to the disease, in the case of normal animals stained vitally they do not do this until after the third or fourth dose of successive daily injections. After many days, nevertheless, the vital stain alone produces enlargement, proliferation, and separation of Kupffer cells so that these are converted into large free phagocytes which may possess one or several nuclei. These are the gigantic macrophages of chronically stained animals. In all our experiments we have used only acutely stained animals, so that the effects of the dye itself are never sufficient to produce the changes. In fact there is no evidence that the dye accentuates the changes appreciably during the time involved in the experiment. The dye, however, shows us the type of the cells entering into the tuberculous granuloma, for when fed to the body fluids in abundance trypan blue finds its way into all cells capable of receiving it. The vital stain is, as it were, a physiological test for the cells. Whatever the fundamental nature of the vital stain produced by trypan blue and the benzidine dyes may be, it is important that this reaction does not occur to any appreciable extent with mononuclear blood cells, and that it does occur emphatically in the case of the hepatic endothelium. By means of this vital test, then, the following phenomena occur when suspensions of tubercle bacilli are let into the portal blood stream. The organisms, swept on by the blood stream, finally lodge in the terminal branches of the portal vein, where they plug the vessels and continue to multiply. They injure the vessel wall and cause around them an exudative inflammatory process, and finally lead to the formation of tubercles situated not only in these areas but also within the liver lobule. The injury to the vessel wall is manifested in the early stages by the presence of vitally stained areas in its structure. The bacteria at the end of half an hour are found to be extracellular in clumps in the larger vessels, but already to some extent in the bodies of vitally stained Kupffer cells throughout the liver. Exudative inflammation manifests itself by the presence of a transitory accumulation of polynuclear leucocytes about the bacterial clumps, which may be seen as early as half an hour after the inoculation. They continue to be present in the larger cell clumps of the periportal areas for many days, but they are rapidly replaced by other cells, mononuclear in type, so that within a day the histological appearance of the portal plug has changed radically. The mononuclear cell thus entering most actively into the reaction is endothelial and not hematogenous in origin, the vital stain enabling us to make a clear distinction. This fact, evident in the portal plugs, is decisively shown in the case of tubercles developing within the liver lobule. Such tubercles probably result from the localization of individual organisms within the Kupffer cells, for the initial stages of such a probable cycle have been found by us. They consist of the occurrence of mitoses in certain Kupffer cells where the Ziehl-Nielson method shows a bacillus or several bacilli to have been phagocytized (figure 4). Rapid growth of the infected cell now takes place, and at thirty-six hours the multinucleated giant cell produced is largely separated from the other endothelium of the vessel wall. Many bacilli exist within the protoplasm of these cells (figure 5), which are especially distinguishable by their intense reaction to the vital stain. They have received trypan blue to such an excess that low power views of liver sections at the thirty-six hour stage show these cells as deep blue spots (figure 8). The origin of the giant cell from the Kupffer cell is evident not only from the above sequence and from the elective stain, but also from the fact that even when fully formed, protoplasmic strands still join it to its mother tissue,—the normal endothelium of the vessel. The strands entangle other cells in their meshes, especially mononuclear blood cells, one of which, of the polyblastic type, has homogeneous protoplasm and is not infrequently encountered in mitosis. These cells are unquestionably of importance in the lesion of tuberculosis. We have seen them abundantly in the capillaries soon after the inoculation and they also occur singly or in nests between the Kupffer cells and liver columns. They are, as a rule, free from the vital dye. They continue to be concerned in the further growth of the tubercle and with the connective tissue cells make the structure of older tubercles relatively complex. On the other hand, little complexity occurs in the structure of the young intralobular masses. The miliary tubercle formed at the end of thirty-six hours is composed of a giant cell, surrounded by epithelioid cells and by blood cells of the above polyblastic type. The giant cell and its so called epithelioid cells are electively stained and are exclusively derived from the hepatic endothelium.