It is a well-known fact that there are discrepancies between titers of the neutralizing antibody in the serum of immunized mice and their protective ability against the intracerebral infection with neurotropic viruses.
In experiments of mice with a fixed strain of the rabies virus, the authors titrated successively the neutralizing antibodies in the serum and the brain tissue and the protective ability against the intracerebral infection following intraperitoneal immunizations with living virus. Antibodies in the brain tissue were measured as follows: mice were thoroughly irrigated with 20ml of 1% citratesaline from the left ventricle of heart after bleeding under ethernarcosis and being cut in the right ventricle of heart. The brain was removed, washed with the saline, and homogenized with broth after weighing. Following centrifugation, the supernatant, after heating at 50°C for 20 minutes, was mixed with the virus suspension, and titrated of its neutralizing property by the intracerebral inoculation to normal mice.
The antibody in the serum was sustained at high titers during 7 weeks after the immunization, whereas that in the brain tissue decreased rather rapidly proportionately to the decrease of protective ability. The seeming parallelism between the latter two factors suggests an important role of the brain tissue antibody in the protection against the intracerebral infection of rabies. virus.

View full abstract

The inhibitory effect of homosulfamin on the intracellular phage growth was investigated here, with respect to the determination of the sensitive part of the phage multiplication course. The latent period of T4r phage was divided, at least, into three distinct stages as to homosulfamin-sensitivity, i.e., the first insensitive stage (first 6 minutes), the second complete sensitive stage (6 to 15 or 16 minutes), and the third or final insensitive stage (16 to 20 minutes), and concluded that this drug may attack some essential reactions relating to either synthesis of virus DNA or the synthesis process of the virus particles from their precursors.
The drug action was analysed from chemical kinetic point of view, and 50 per cent inhibition does on bacterial and virus growth were 0.18 and 0.29 per cent, respectively.

View full abstract

The mode of bacterial responses against a temperate phage infection was investigated with respect to the physiological conditions of the host bacterial cell. β [A34] phage -infected Komagome A strain (Shigella flexineri type 5) showed 4 response types, i.e., productive-, reversible-, lethal-, and lysogenic responses, and these fraction were affected by the bacterial conditions as follows:
And concluded the establishment of phage multiplication process was controlled markedly by the host cell whereas that of lysogenization process was relatively unaffected by the physiological conditions of the host suggesting that lysogenization process was due to the heterogeneity of the phage particles.

View full abstract

Similarities in various characteristics between the ectromelia and the vaccinia viruses pointed out by Burnet and Fenner were also re-established by the authors experiments. However, it was revealed that, while mice immunized with live vaccinia virus resisted the intraperitoneal infection of the ectromelia (“Hampstead” strain), they showed same local lesions as the control animals when challenged with the ectromelia virus plantarly. On the other hand, mice surviving from the ectromelia infection could perfectly get rid of even the plantar reinfection.
Similar relations concerning the injection route are observed also in the neutralization test. Thus, although the ectromelia-neutralizing ability of anti-vaccinia or anti-ectromelia mouse sera could be demonstrated when the virus-serum mixture was injected intraperitoneally into mice, it was not observed when the mixture was injected plantarly.
The authors are tempted to assume the necessity that the systemic general infection and the local disease must be considered separately each other in the study of mouse ectromelia.
An epidemic of ectromelia, from which a new virus strain, “Tama”, was isolated, was observed and investigated in a large group of mice purchased from an animal dealer in Tokyo.

View full abstract

The partial purification of influenza virus was recognized to be given to a certain degree, by absorption and elution using such absorbents as ion-exchange resins, clay, kaoline, diatom earth, or chicken red cells. Furthermore, purification was obtained in considerably high degree, by combining precipitation method to the materials, preliminarily purified by these absorbents, or by combining methanol precipitation method as well as a few absorbents.
In a experiment in which methanol precipitation method was combined with the purification procedure by chicken red cells, applied to the supernatant fluid obtained by the absorption to and elution from ionexchange resin, the following result was obtained: In the final purified material, the concentration of the virus, designated by the hemagglutination titer, increased by 4 times and the nitogen content decreased to 99.4% compared with those of the original material.

View full abstract

Bacterial virus strain A active against Pseudomonas solanacearum E. F. Smith strain B19 was purified by the differential centrifugation at 26, 000g for 60 min., 59, 000g 60 min., and 20, 000g 120 min. According to the electron-microscopic observations, the purified virus appeared uniformly as the spherical particles approximately 70-80mμ in diameter (Fig. 1). The surface of virus particle was not smooth, and assumed to have some internal structures within the particle.
The cell contents of host bacteria, infected with the virus, were filled with very small protoplasmic particles and the new virus particles. After the latent period, the membranes of the infected bacterial cells were strongly ruptured and the proliferated new virus particles were liberated from the bursting cells (Fig. 3).

View full abstract

In January, 1953, there was an outbreak of swine influenza-like disease in Shizuoka Prefecture, and in July, 1953, another outbreak of the same disease in our institute.
In our laboratory, the virus was isolated from three materials of lungs with haemorrhagic lesions and consolidation by conducting intranasal inoculation into mice, and from another two materials by practicing amniotic method. As regards the elevation of antibody in a swine serum against the virus, the complement-fixation reaction and the haemagglutination-inhibition test were proved effective.
Our finding of the swine inoculated this virus is as follows: Observed were pathologically, oedem and haemorrhage of lung, follicle swelling of spleen, haemorrhage of urine bladder, swelling and congestion of each lymphnodes, and chinically, slight rise of temperature, (40-50.5°C), chilliness, decrease of appetite and coughing. Beside this, what were observed in circulating blood are such as the appearance of myelocyte and metamyelocyte, and the increase of rod nuclear cells in neutrophiles and of monocyte.
As a result of making tests in vitro and vivo on the newly isolated virus, we learned of its main properties as follows:
1. All the mice inoculated the virus through intranasal route, died after 4 to 8 days, presenting such a symptom as consolidation of lung, similar to that of influenza. Microscopically observed was catarrharic broach-pneumonia with severe infiltration of neutrophile leucocytes and necrosis of bronchial epithelcell. Virus multiplication was observed in spleen, lung and lymphnode.
2. As to the isolation of the virus from the swine, the amniotic method was successful, while the allantoic method was not. In case the virus was demonstrated by haemagglutination in the second or third serial amniotic passage, the cultivation of the virus in allantoic cavity and yolk sack of embryo could be made possible and the serial passage of the virus, made easy to develop through any route.
3. When the virus was inoculated into yolk sack of 6 to 8 day old embryo, the embryo, as a rule, was killed 48 hours after the inoculation; but it survived when the virus was inoculated on chorioallantoic membrane or in allantoic cavity.
Regardless of its route, the virus developed positively in 48 to 96 hours after the inoculation.
4. The virus titer in allantoic fluid usually indicated 10^-5 to 10^-7 and haemagglutination titer, 1: 1, 000-1:2, 000. And the haemagglutinative potency of the virus was demonstrated in the red cells of fowl, guinea-pig, sheep, goat, pig, mouse, pigeon, human (o type), rabbit and cattle, but it turned out negative in the red cell of horse.
5. The virus was completely adsorbed into fowl red cell at 4°C in 30 minutes and was eluted at 22°C in 10 hours.
6. The haemagglutinin was destroyed by heating at 50°C in 5 minutes, while the infectivity was entirely destroyed at 22°C in 30 days, and by heating at 50°C in 10 minutes.
7. In the process of isolation of the swine virus, the variation of phases “O” to “D”, which was described by Dr. Burnet et al. in the process of isolation of human influenza A virus, was observed. Outline of the variation of strain S was schematically shown in Chart No. 8. Also, in the case of isolation of strain 153, HME and 3471, the similar phenomenon was observed.
8. The filtrability of the virus, going through Seitz EK, Berkefeld w. and Chamberland L3, was proved.

View full abstract

In view of the recent progress in the study of hemagglutination by neurotropic viruses, it is necessary to exert caution regarding the non-specific hemagglutinin of normal mouse brain. In my studies on Japanese B encephalitis virus hemagglutinin it was observed the hen erythrocytes were agglutinated by normal mouse brain under condition to be described.
In 1949 Olitsky & Yager recognized some non-specifichemagglutination in normal and Columbia-SK infected mouse brain, but many of these non-specific hemagglutinins including the presently studied hen red cell hemagglutinin did not ocur constantly and for this reason the properties of the hen red cell hemagglutinin were investigated and compared with those of the specific hemagglutinin of Japanese B encephalitis virus.
The hen red cell hemagglutinin of normal and infected (J. B. E., St. Louis or Columbia-SK) mouse brain could be recognized after along reaction-time. Thus, if the erythrocytes pattern was negative following the first sedimentation, subsequent reshaking and sedimentation for one to two times would certainly result in hemagglutination to a titer or 1:100 to 1:200. The best results are obtained at ph 6.8 to 7.5 at room temperature (under 20°C). The non-specific hemagglutinin was inhibited by normal sera (either untreated or treated with chloroform) and skimmed milk. This hemagglutinin was not inactivated by trypsin or formalin at low temperature but almost completely removed by treatment with lipid solvent such as chloroform at low temperature. Specific Japanese B encephalitis virus hemagglutinin was distinguished by being inactivated by the former treatment but was unaffected by the later treatment.
The possibility that the non-specific lipid hemagglutinin in normal mouse brain for hen red cell and the non-specific lipid hemagglutinin for fowl's red cell described by Stone in 1946 are similar was discussed.

View full abstract

For the production of highly specific complement fixing antigen of neurotropic viruses the extraction of non-specific substance by lipid solvents has often been used. These extraction procedures were investigated for their effect upon the hemagglutinin (for chick erythrocyte) of Japanese B encephalitis virus. Extraction of mouse brain was performed by 11 different lipid solvents: petroleum ether, ligroine, ethylether, amylalcohol, toluene, pyrigine, acetone, benzene, chloroform, carbon disulfide, and carbon tetrachloride. When extraction were carried out at low temperature (not above 10°C) either with or without previous lyophilization of the virus suspension, the difference between the specific Japanese B encephalitis virus hemagglutinin and the non-specific hen red cell hemagglutinin were as previously reported.
Specifically, on successive extraction by benzene, chloroform, acetone, and ethylether after lyophilization the non-specific hen red cell hemagglutinin was almost completely removed while the specific Japanese B encephalitis virus hemagglutinin and the specific complement fixing antigen were decreased only slightly inpotency. In this extracted samples with a relatively high specific hemagglutinin titer but without significant amounts of non-specific hemagglutinin (for hen red cell), inactivation of the specific hemagglutinin was attempted by treatment with carbolic acid, formalin, and trypsin. In addition by chick erythrocytes was tried. While specific Japanese B encephalitis virus hemagglutinin was inactivated by the above mentioned reagents and adsorbed by chick red cells, the complement fixing antigen potency was undiminished. Extraction by toluene resulted in a similar decrease in specific hemagglutinin titer without effecting complement fixing titer. Direct extraction of virus suspension by chloroform had same effect as the successive extraction described above. Ultracentrifugation of this sample at 23, 000r.p.m. (average g: 33, 000) resulted in almost complete sedimentation of the specific hemagglutinin, while the complement fixing antigen remained almost undiminished in the supernatant fluid.
These experiments confirm the difference between the specific hemagglutinin of Tapanese B encephalitis virus and the non-specific hen erythrocyte hemagglutinin, and present evidence that the specific hemagglutinin, and the complement fixing antigen are seperate entities.

View full abstract

Normal cow serum gamma globulin (exactly, γ-globulin and β-globulin fraction) was fractionated by the low temperature ethanol method. Experiments on the control and treatment of poliomyelitis infection (of Lansing strain) was carried out, using mouse as experimental animal. The results were as follows.
Gamma globulin of normal cow serum was effective, when administered for the purpose of the prevention of the disease.
The influence or effectivity of γ-globulin administration depends largely on the quantitative difference of the titers of the neutralizing antibody against the virus.
To start γ-globulin administration shortly before the virus challenge seems to be most effective to satisfy the preventive purpose.
As for the treatment after the virus invasion, the mouse group which had been passively immunized with γ-globulin before the time of the virus challenge was divided into two groups, one of which had received the second treatment of γ-globulin, while the other group did not. The group that had the second treatment showed the prolongation of the incubation period and also the decrease in mortality, compared with the other group and the non-treated group. Based on the present experiment, it may be said that not only the prevention but also the therapy of poliomyelitis infection is not without hope, if adequate γ-globulin is properly administered.

View full abstract