In the previous paper the authors reported on the possibility of so called local tissue immunity of guinea-pigs inoculated with the virus of Japanese B encephalitis. After that the authors could demonstrate the local concentration of neutralizing antibody in the brain tissue of the animal immunized intracerebrally with the active or inactivated virus. In the animals immunized subcutaneously, and particularly in those immunized passively with homologous immune serum, no antibody concentration could be confirmed.
The intracerebral antibody concentration of this kind may depend upon the intracerebral local antibody production or upon the increase of permeability of the so called blood liquor barrier of brain dueing to the intracerebral inoculation. The both may be possible and await the further scrutiny in the future.

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Most of the inactivated chicken sera immune to the virus of Japanese encephalitis do not fix complement by the usual technic, and unheated chicken sera have anticomplementary property so strong that specific complement fixation test can not be obtained.
Complement fixation inhibition test is successfully applied to the chicken sera using horse immune serum as an indicator serum.
When the immune chicken sera heated at 45°-75°C for 30 minutes were tested by common complement fixation and complement fixation inhibition test, in the former the reaction was heat-labile, and in the latter that was heat-stable remarkably.
Testing chicken sera immune to the influenza virus (PR 8, FM 1), the results of the reaction resembled extremely to that of Japanese encephalitis virus.

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Up to this time all the complement fixation inhibition tests reprted by many workers i.e. Rice, Hilleman, Wolfe, Downie et al. were based on the serum dilution method, but our method described in this paper was carried out by the method of complement dilutioon, modifing the Nakamura's technic of C-F-T run on bovine sera with antigen prepared from rinder-pest virus.
In this experiments guinea pigs sera obtained by immunization with guinea pig adapted fowl plague virus (infected brain) were employed as complement fixing sera. And complement fixation inhibition tests (C-F-I-T) were successfully applied to chiken sera obtained by immunization with the attenuated virus, using the guinea pig sera.
In this complement dilution method positive results were obtained even though anti-complemetary properties of sera and antigens or Forsman's and normal tissue antibodies were present.

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As the results of a prelininary test it was found that the specificity in the direct complement fixation test run on pigeon sera obtained by immunization with the pigeon adapted newcastle disease virus (infected pigeon brain) was observed by the complement dilution method described already in our previous paper.
In this experiment the antigen was prepared from infected embryo and chorioallantoic fluid purified by simple centrifugation (4000 r.p.m. 20 minutes). Then using the pigeon sera and the antigen, specific C-F-I-T was carried out on chicken sera obtained by the vaccination with the living attenuated virus.
As regard to the results of these tests it was found that the complement fixation inhibitory antibodies were not always paralleled to that of hemagglutination inhibitory antibodies, and that the former was demonstrated generally earlier than the latter.
The fowl plague virus were distinguished clearly from the newcastle disease virus by means of the C-F-T, using the method of the direct complement dilution.

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A further observation of inclusion bodies in monocytes drawn out from the peritoneal cavity at varied intervals after intraperitoneal inoculation of ectromelia virus was made availing phase contrast microscope. A. 24 hours after the inoculation, young inclusion bodies appeared distinctly as round or oval ones in slightly deeper tone than the surrounding cytoplasm. Elementary bodies with the definite size were not distributed inside these inclusion bodies. This fact shows that in early stage two components of the inclusion body, namely, the matrix and elementary bodies develop from different bases, the former from cellular constituent, while the latter only remaing on the surface of this matrix. In the following stages, elementary bodies begin to invade the matrix membrane and subsequently fill up the inclusion body.
B. From this new finding, we were obliged to seek the initial structure for these inclusion bodies. As the young inclusion bodies located in general amidst the mitochondrial zone more or less at the periphery of the cytoplasm, indifferent from the Golgi zone, so we set focus to the relationship between the initial inclusion body and mitochondrias. The initial form found 5 to 6 hours after the virus inoculation starts from the thickening of one or several agglomerated mitochondrias, which develop after 10 to 12 hours as a distinct form of the inclusion body.
Observing the various modi of infected tissues, we can classify two types of the cellular infection; one, like liver cells, being damaged till to rapid necrosis without formation of inclusion body in general, whereas the other for instance monocytes and epidermis, being relative resistant with the formation of the inclusion body in the same animal. The auther is tempted to consider the inclusion as a buffer structure developed from the cellular organella against virus.

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For about two months from March, 1952, seventeen newborns suffered from one to another of the following symptoms and eleven out of them died. The clinical characteristics of this pneumonitis of newborns were as follows: high contagiousness, high mortality up to 64.7%, slight symptoms in the premature infants, sudden onset without any prodromal symptoms; high fever (38.5-41.8°C); respiratory distress, cyanosis, increased jaundice, rhinopharyngitis, sweating, marked leucocytosis (20, 000-40, 000), especially due to neutrophilia (60-80%) from the beginning of the illness, high grade of anemia in the reconvalescence stadium; slight pathological findings on the chest, though diffuse shadows were obviously found in the lung fields on the roentgenogram; and no inclusion bodies showed in throat smears. Both the Hirst- and cold hemagglutination test were negative. All kinds of antibiotics were ineffective.
This newborn virus pneumonitis is diagnosed differentially from the pneumonitis described by Nitschke, Adams and others. This pneumonitis can be clinically concluded to be a new type of newborn pneumonitis and is termed as “Newborn Virus Pneumonitis (Type Sendai)”.

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In the spring of 1952, an outbreak of newborn Pneumonitis occurred in this University Hospital.
In our laboratory, the isolation of virus from 5 autopsic materials was carried out by nasal instillation of lung suspensions to mice. All of the mice died between 3 to 7 days and presented a consolidation of lungs, resembling that of influenza. The pathological findings of lungs were similar to that of newbornes and no inclusion bodies have been smeared. Except pernasal instillation also intracerebral and intraperitoneal injection to mice caused the death of mice and consolidation of lungs.
After adaptation to mice, they could be cultivated in amniotic and allantoic cavity of fertile eggs, and after subsequent passage, hemagglutinin (HA) was proved in both cavities. HA was adsorbed to and eluted from chicken erythrocytes. Heat stability of HA was so feeble when compared to another virus, belonging to MNI. Receptor gradient of the virus was examined and it was arranged between NDV and FMI.
HA-inhibition test was performed by several specimens of monovalent rooster serum, i.e. of this virus (Fushimi), Mumps (Enders), Newcastle (Fukumi), Influenza A (PR8, WS), A' (FMI, Lépine and Matsumoto), B (Lee, Seattle and T14) and C (1233). No other serum could inhibit this HA, except the correspondent serum of the agent. Beside this Akitsugu and Tochigi strains, both isolated from adult's influenza in other laboratories were proved to possess the same antigenic structure with this virus. The neutralization antibody was proved with convalescent serum of patient and immun serum of rabbit.
Recently we could find again a new sporadic case, and isolated the same virus from throat and the elevation of antibody against this virus in paired serum of this patient was proved by HA-inhibition. test. Further research proved the enzootic nature of this virus i.e. the same virus was isolated from normal laboratory mice. But all of mice examined have no antibody against this virus.
Intranasal instillation of the virus to a volunteer caused a fever of the saddleback type and high elevation of antibody. The virus was also recovered from throat 2 times after onset.
The purification studies of the virus by the supercentrifuge revealed a spherical particle 150 to 200mμ in size resembling rather to mumps virus and NDV than influenza virus.

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This is to report on the 9 autopsy cases clinically observed by Sano et al. Grossly, in all cases homorrhagic pneumonia was observed in both sides and in all lobes of the lung, showing the appearance of socalled splenization. No pleurisy was observed. Bleeding was found the serosa of all bodily cavities, in the mucosa of alimentary canals and in the retroperitoneum etc. The leptomeninx was congested and slightly turbid, in 4 cases the spleens were slightly enlarged and in 7, we observed hemorrhagec erosion in the stomachs.
Histological examinations: In the affected part of the lung the alveolar septa were often greatly distended owing to excessive blood congestion and cellular infiltration, giving the appearance of interalveolitis. The infilitrating cells mainly consisted of medium-sized mononuclear cells and cells with oblong nuelei, mingled with a small number of leucocytes. Edema, atelectasis and emphysema were found scattered in disorder among the area of hemorrhages and interalveoritis. In some part desquamation of the bronchiolar epithelium was visible, but no necrotic and degenerative change was found, except a case in which klebsiella pneumoniae was discovered from the lung. No hyaline membrane was found on the surface of alveoli and bronchioli, nor any hyaline thrombus in the capillaries. Fibrin and swallowed epithelium were occasionally found in the air sacs. In spite of hard investigation, no inclusion bodies could in the bronchial and alvleolar epitheliums. Beside the above, hepatitis, serous hepatitis, light leptomeningitis and occasionally nephrosis were observed.

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About one year later from the epidimic outbreak of Newborn Pneumonitis (Type Sendai) in this hospital, we experienced again a sporadic case in March 1953. The patient, 9 days old male, was diagnosed as one of this type pneumonia by clinical symptoms.
The isolation of virus from throat washings was then attended by intranasal instillation to mice. The mice died representing typical consolidation of lungs and after 2 times passage in mice, the virus was cultivated in both amniotic and allantoic cavity of fertile eggs. The antigenic structure of this virus examined by hemagglutination inhibtion test was quite similar with that of Fushimi strain, isolated in the last year.
Moreover the elevation of hemagglutinin-inhibiting antibody in this patient using Fushimi and BE₂ (isolated from normal mice) strains as antigen was ascertained. The fact was further proved by neutralization test in fertile eggs.
In conclusion, it was revealed, that the pathogenicity of this virus against newbornes was consistently proved, although these group viruses may be isolated from normal mice.

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