Hospital of The Rockefeller Institute for Medical Research.
J Exp Med. 1931 Mar 31;53(4):471-92. doi: 10.1084/jem.53.4.471.
During the course of a study of different strains of influenza bacilli, fifteen strains have been found to form colonies of a different appearance from that usually considered typical of influenza bacilli. These colonies are smooth, more opaque, and are iridescent in oblique transmitted light. Most of these strains were isolated from patients in whom these organisms seemed to play a pathogenic rô1e. When these strains were grown repeatedly on blood agar, other colonies appeared which were smaller, less smooth, less opaque, and not iridescent, and when subcultures were made from these rough colonies, all of the colonies were of this character. Further study of the cultures obtained from these smooth and rough colonies have shown that one is a variant of the other. The strain from the smooth colony has been called an S strain, that from the rough colony an R strain. Certain differences in the morphology of the organisms in the R and S strains have been observed. Of special importance is the fact that the bacteria of the S strains are possessed of capsules. It has also been found that the S strains are somewhat more virulent for laboratory animals than are the R strains. In the supernatant fluid of broth cultures of S strains, and in the washing fluid of S bacteria grown on agar, there is present a soluble substance which, in the presence of homologous immune serum, gives rise to a precipitate. No reaction of this kind, however, occurs with the cultures of the R strains. By means of cross precipitin reactions, employing antisera against the different S strains, it has been found that the fifteen strains studied may be divided into two distinct immunological groups. Three of these strains belong in one group, Type a, and the remaining twelve in another group, Type b. Seven of the strains studied were isolated from the spinal fluid in cases of meningitis, and all of these strains are of Type b. Agglutination tests performed at 37 degrees C. with these fifteen S strains have revealed the same specific type relationships among the organisms as did the precipitin tests. The R strains on the other hand, exhibit no similar type agglutinations. If the agglutination tests are made at a higher temperature, 47 degrees C., the S strains also fail to show the specific type reactions which occur at 37 degrees C. Certain differences between other biochemical reactions exhibited by the two types of strains have been noted, but it is not believed that they are sufficiently constant to be of great significance. When S strains are grown on artificial media outside the animal body, they tend to be converted into the R form. The rapidity and the readiness with which this conversion occurs depend on certain conditions, such as the kind of media employed, the temperature at which the cultures are kept, and the atmospheric conditions under which they are cultivated. The rate of conversion is increased when the S strains are grown in media containing anti-S immune serum of the homologous type. On the other hand, conversion of R strains into the S form occurs with much less readiness, and then only if very particular conditions are present. On one occasion conversion occurred when an R strain was grown in a medium containing anti-R immune serum. On two other occasions this same strain changed from the R to the S form during passage through animals. With other R strains it has so far been impossible to bring about this transformation. These studies indicate that the bacteria belonging in the group Hemophilus influenzae exhibit changes in pathogenicity and immunological specificity, which are analogous to those shown by the bacteria of the pneumococcus group. It is important to continue this study, with the technique which has been developed, to include a much larger number of strains. On account of the readiness with which the S strains of influenza bacilli lose their type specificity when grown on artificial culture media, it is important that the organisms be studied as soon as possible after removal from their pathological sources. It is not impossible that many strains lose their specificity immediately after removal from the host, and that the specific immunological differentiation of many strains may, for that reason, be very difficult, if not impossible.
在研究不同流感杆菌菌株的过程中，发现有 15 株形成的菌落与通常认为的流感杆菌典型菌落不同。这些菌落光滑、更不透明，在斜射光下呈虹彩。这些菌株大多数是从似乎具有致病性的患者中分离出来的。当这些菌株在血琼脂上反复生长时，会出现其他更小、更粗糙、不透明、无虹彩的菌落，而从这些粗糙菌落进行亚培养时，所有的菌落都是这种特征。进一步研究从这些光滑和粗糙菌落中获得的培养物表明，一种是另一种的变体。来自光滑菌落的菌株被称为 S 株，来自粗糙菌落的菌株被称为 R 株。已经观察到 R 和 S 菌株中生物体的形态存在某些差异。特别重要的是，S 菌株的细菌具有荚膜。还发现 S 菌株对实验动物的毒力比 R 菌株略高。在 S 菌株肉汤培养物的上清液中，以及在琼脂上生长的 S 细菌的洗涤液中，存在一种可溶性物质，在同源免疫血清存在的情况下，会产生沉淀。然而，用 R 菌株的培养物不会发生这种反应。通过交叉沉淀反应，用针对不同 S 株的抗血清，发现研究的 15 株菌株可以分为两个不同的免疫组。其中三株属于一组，A型，其余 12 株属于另一组，B 型。研究的 7 株菌株是从脑膜炎患者的脊髓液中分离出来的，所有这些菌株均为 B 型。在 37°C 进行的凝集试验表明，这些 15 株 S 株之间的生物体具有与沉淀试验相同的特定类型关系。另一方面，R 株没有表现出类似的类型凝集。如果在较高的温度 47°C 下进行凝集试验，S 株也不会显示在 37°C 下发生的特定类型反应。已经注意到两种类型的菌株表现出的某些生化反应之间的差异，但据信它们不够稳定，没有太大意义。当 S 株在动物体外的人工培养基上生长时，它们往往会转化为 R 形式。这种转化的速度和准备程度取决于某些条件，例如所使用的培养基类型、培养物保持的温度以及培养物所处的大气条件。当 S 株在含有同源型抗 S 免疫血清的培养基中生长时，转化率会增加。另一方面，R 株转化为 S 株的速度较慢，只有在特定条件下才会发生。有一次，当 R 株在含有抗 R 免疫血清的培养基中生长时，这种转化就发生了。在另外两次，同一种菌株在通过动物时从 R 型转变为 S 型。对于其他 R 株，目前还不可能实现这种转化。这些研究表明，属于流感嗜血杆菌组的细菌表现出与肺炎球菌组细菌相似的致病性和免疫特异性变化。使用已经开发的技术继续这项研究，包括更多数量的菌株非常重要。由于流感杆菌的 S 株在人工培养物上生长时很容易失去其类型特异性，因此重要的是尽快从其病理来源中研究这些生物体。许多菌株在从宿主中取出后立即失去特异性并非不可能，许多菌株的特异性免疫分化可能因此非常困难，如果不是不可能的话。
