In order to study on the relation of vaccinial infection to its immunity which is induced by the former, the changes at the site of rabbit skin inoculated with vaccinia virus were investigated by pathological, immunological and biological means, and the relationship among various factors concerning the infection was pursued. The findings obtained are summarized as follows.
1. Vaccinia virus multiplied at the inoculated site of rabbit skin prior to the appearance of the pathologic lesions, reached the maximum with the latter almost at the same time, and then tended to decrease previously to the lesions.
2. The complement fixing antigen being produced at the infected site increased parallel with the viral multiplication. It, however, reached the maximum somewhat later and kept its maximum titer a little while longer.
3. The antigenicity of the extract of the infected skin related to the degree of the pathologic changes of the skin; namely, in the cases of the weak lesion the antigenicity in vitro was lower than that, in the cases of the intensive one. (See P. 97)
4. The proliferation of epithelium accompanied the increase of ribonucleic acid simultaneously.
5. The virus neutralizing activity of the extract of the infected skin riled parallel with the increase of rγglobulin in the extract, which was electrophoretically observed as the fraction IV. It seemed therefore that the virus neutralizing antibody was contained in the fraction IV of the extract of the infected skin.

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It is well known that Japanese B encephalitis is a mosquito borne disease. Some Japanese investigators, however, have an idea that the alimentary canal may be an entry of Japanese B encephlitis virus similar to that in poliomyelitis virus. Hence the attempt was made to confirm whether or not the disease may develop following oral, intestinal al or rectal administration of the virus to hamsters.
Each of 2 hamsters was orally given two infected mouse brains. Out of these hamsters 6 (19%) got disease. Four hamsters each orally given five infected mouse brains were followed up as for the distribution of the virus in each organ (blood, liver, peritoneum, spleen, mesenterial lymphnodules, bulbus olfactorius, cerebrum, cerebellum and spinal cord), and the similar results to those following nasal infection in mouse were obtained.
Four hamsters had no encephalitis following direct injection of 0.3cc of 50% infected brain emulsion into surgically operated intestinal canal.
Out of 36 hamsters rectally injected with 5cc of 10% infected mouse brain emulsion without stopping anus with nippers, 12 hamsters (33%) got the disease. Whereas none out of 4 hamsters rectally injected with 5cc of 20% infected mouse brain emulsion stopping the anus with nippers for 24 hours and washed with acetic acid solution (pH 1.0) got the disease. The virus distribution in each organ was followed up daily after rectal injection of virus in the same condition as the above mentioned experiments and the virus was confirmed only in the contents of intestines, intestinal wall and blood in 2 hours after administration.
From the forgoing experiments it may be concluded that there is no evidence for the entry of the Japanese B encephalitis virus from alimentary canal into central nervous system in hamsters.
Four monkeys were orally given 6 mouse brains to each, and no symptoms indicating the onset of the disease were recognized. The viruses were not recovered from blood and feces. Rise of neither neutralising nor complement fixing antibody titers was not recognized.

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Histological changes of the skin inoculated with vaccine virus by means of the sand paper method are observed fromimmune biological point of view. (See P. 108)
1) After 5 hours of this vaccination, epidermis cell changes to necrosis and the neutrophilic infiltration appears in demarcation zone. When these tissues discharged after 5 days, there prepared already regenerated epidermis beneath the scab. However, these changes are seen in the contrast experiment by simple sand paper rabbing or electric burning. So the charactericity of the vaccinated epidermis should be represented by the regenerated epidermis in the following stage.
2) The inclusion body appears in the cell body of the regenerated hypertrophic epidermis and the follicule epithelium after 2 days and till 7th day. After 4 days a part of the hypertrophic epidermis is liquidated calling the infiltration of the neutrophilic cell and postulating the locus. Following this reaction the monocytic infiltration appears in the papilla, growing gradually to the depth of the corium along the hair follicule and reaching to the reticular layer.
3) After 8 days the hemorrhagic inflammation appears in the monocyte infiltrated corium which makes this leisions necrotic and causes to fall off. After this time neither the inclusion bodies nor the monocytic infiltration were to be found. These findings show that there is a close correlation between the existence of the inclusion body and the monocytic reaction in the epidermis. It can be said that the tissue reaction against the vaccinia virus is the monocytic nature.
4) The colour of the neutral-red vacuoles of the histiocyte in the reticular layer was observed supravitall, which changed to brownish during the period of 3 to 17 days after the vaccination in parallel to the alkalosis which occured the inoculated tissue.
5) The plasma cells began to grow close to the addventitia cells in the subcutaneous tissue 4 days after the inoculation. After 7 days they infiltrate in cluster and after 8 days explosively around the area of the hemorrhagic infiltration. The proliferation of the plasma cell occurs on the period of alkalosis of the inoculated locus as well as the bone marrow successive to the leucocyte infiltration and alkalosis of the tissues.
6) 20 to 25 days after the primary vaccination the reinfection experiment was applied to the abdominal side of the animals (for the observation of the Pirquet's immediate reaction). The erythema and the swelling of the inoculated locus reached to its maximum after 24 hours transitorily without any following pustulation.
7) The neutrophilic reaction on the vaccinated locus at the beginning of the second inoculation appeared more intense than that of the primary, somewhat like the mode of Arthus Phenomenon.
8) As the destruction of the virus was performed by this Phenomenon so neither the inclusion body nor the monocytic infiltration was to be seen in this reinoculation. However, the call of the plasma cell reaction is rather quickened.
9) pH of the locus is alkalified in parallel to its erythema and then returned to the normal state in a short time. However, the brown colour of histiocyte-vacuoles is still remained after the recovery of the tissue pH, which means brown colour of histiocyte-vacuoles is not influenced simply by the tissue pH.

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Le pouvoir neutralisant d'un échatitillon d'immunsérum a été évalué au moyen d'un matériel virulent lyophilisé et d'une émulsion virulente fraîchement préparée. Les mélanges virus-serums ont été laissés à 2-3° pendant 0, 24 ou 48 heures avant l'injection intradermique. Le titre du sérum déterminé an moyen d'un matériel virulent lyophilisé. s'est montré supérieur à celui évalué par une émulsion non traitée.
Les virus inactifs que le matériel pourrait contenir en certaine quantité, seraient-ils capables d'accélérer la neutralisation des virus actifs par l'immunsérum? Afin d'élucider ce point, nous avons essayé d'utiliser un matériel virulent partiellement inactivé par des rayons ultraviolets pour pratiquer la réaction de neutralisation. Les résulats nous out iudiqué que le matériel partiellement inactivé se comporte dans la reaction de neutralisation a la manière analogue au matériel virulent lyophilisé.
Pour confirmer le fait que des virus inactivés par irradiation peuvent accélérer la neutralisation de virus actifs, nous aeons ajouté l'émulsion complètement inactivée aux mélanges virus-sérums. Le taux de neutralisation s'est élevé en présence de virus inactifs.
Dans le but de déterminer si les virus inactifs absorbent des anticorps protecteurs, les mélanges de l'immunsérum et de virus inactifs, après une incubation de 24 heures, ont été filtrés sur filtres de Seitz. On a évalué le pouvoir neutralisant des filtrats obtenus. Dans ces conditions, les virus inactifs ont absorbé les anticorps protecteurs. (See P. 119)
Conclusion: Le virus vaccinal inactivé par irradiation ultraviolette possède la propriété d'accélérer la neutralisation du virus actif par l'immunsérum ainsi que la propriété d'ahsorber les anticorps neutralisants.

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Neutralization reaction for the potency measurement of the influenza vaccine has so far been carried out by means of mouse inoculation procedure, but this involves some disadvantages, namely first, sometimes non-specific lesions are encountered with in the lungs of mice autopsied and disturbs correct judgement of infective endopoint, second, if newly isolated virus strains would be employed for production and consequently potency measurement of vaccine, it might be difficult to use mice for potency measurement because most of newly isolated strains of influenza virus require more or less time to be adapted to mice, and so on. In these reason experiments were made to obtaine some data necessary for carrying out potency measurement of vaccine with eggs. (Refer to Pp. 123-125)

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Upon studying the relationship between various kinds of wild birds and Japanese B Encephalitis virus, it is revealed that night herons present especially high positive rate of J. B. E. neutralizing antibody.
Consequently, in 1952, test for the presence of viremia, isolation of virus from bird brains, neutralization test and CFT (complement fixation test) of the sera of herons and tattlers were conducted.
O 23 samples of heron sera 8 samples (34%) were neutralizing antibody positive and all negative for CFT. While, 34 samples of tattler sera showed all neutralizing antibody negative, and only 4 samples of them were compelement fixing antibody positive.
Presence of viremia in 16 herons and tattlers were not recognized. Isolation of the virus from the brains of 40 herons and tattlers, and also from the parasites of herons were tried, but all in negative results.

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We tried to make clear the mode of action of HCl-Cysteine and Glutamic acid on the phage-hostcell system, and gained several interesting results. (See P. 130)
1) HCl-Cysteine inhibit a reaction necessary for the production of active phages. Inhibition of Virus growth was obtained when HCl-Cysteine added up to 8 to 11 minutes after infections Removal of HCl-Cysteine from the phage infected coli early in the course of latent period, namely 8 minutes, permit phage liberation. And the burst sizes of phage decreased progressively with lengthning of the interval between infection and removal of HCl-Cysteine.
When removal of HCl-Cysteine from the phage host cell system 14 to 25 minutes after infection, the liberation of phage was found and burst size of phage were as large as control.
2) In nephelometric examination, HCl-Cysteine failed to inhibit lysis of phage infected host cell, but HCl-Cysteine M/6000/ml and M/3000/ml delayed the beginnings of lysis curves.
In these lysis increasing of phage were could not found.
3) Glutamic acid or Aspartic acid was found to prevent HCl-Cysteine inhibition of phage multiplication. Precultivation of host cell in synthetic medium countaining Glutamic acid or Aspartic acid permits phage multiplication to occure in spite of HCl-Cysteine.
And these cases the burst sizes of phage were about 1/2 compaire with control.
While phage, HCl-Cysteine and Glutamic acid (or Aspartic acid) added simultaneously to the culture of host cell in synthetic medium, the burst sizes were smaller than above mentioned results.
DL-Tryptophane or L-Methionine could not prevent the inhibition of phage multiplication by HCl-Cysteine.

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