Since the work of Orgega & Mellors the indirect method of fluoresceint antibody technique has been exclusively used for the demonstration of nephrotoxin in vivo. As far as the immun reaction in vivo is concerned, however, the analysis using the intravenousinjection of fluorescen labeled nephrotoxin should be the first choice. After preparing highly specific F.I.T.C. labeled nephrotoxic rabbit gamma-glubulin with F/P molar ratio fast equal to 1.0., we perofrmed several experiments on the nephrotoxic nephritis injecting these antibodies intravenously. The most remarkable finding by this method is that specific fluorescence in glomeruli localizes only in porper basement membrane compared to the indirect method (staining with fluorescen anti-rabbit gamma-globulin), by which specific fiuorescein are recognized not only in basement membrane proper, but also in epithlial and endothelial structures. On the other hand, an in vitro staining of normal rat kidney with this labeled nephrotoxic antibodies shows that specific fluorescence distributes in Bowman's capsules, tubular cytoplasm, its basement membrane and small blood vessels as well as glomerular capillary loops. It is a very striking fact that the same antibodies react in quite different way in vivo and in vitro. These data suggest that the so-called renal tissue antigenicity and that of inducing nephrotoxic nephritis in vivo is an essentially different problem. Furthermore, following experiments were performed. (I) The application of our method to the study of purification of effective nephrotoxic antigens. (2) The very short term (within 20 minutes) and long term (till 8 weeks) observation of specificfluorescence in gloeruli after injection of labeled nephrotoxin.

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Repository injection method has recently developed in the field of hyposensitization therapy, which aimed tohave an advantage over the conventional aqueous antigen method in saving time of troublesome multiple injections and also in increasing the effect of the therapy. This report was designed to check whether "Drakeol" (mineral oil) or "Arlacel A" (emulsifying agent), two constuitents of adjuvant often used in this method, produce any pathological changes in the parenchymal oragans. Twenty-nine mice were given "Drakeol", "Arlacel A", or both, intraperitoneally, for 6-12 weeks, and histological changes of the liver and kidney were examined. Absorption of these substances from the peritoneal cavity proved good; no visible remainder was seen in any mice when they were sacrificed. 0.05 ml. of Drakeol and 0.05 ml. of Arlacel, given once a week, could not induce any pathological changes both in the liver and in the kidney after 6 weeks' administration. In case of repository injection, if they are employedonce or twice in the form of emulsified antigen of 0.5-1.0 ml. at most the total amount of the individual adjuvant constituents injected into the human body is estimated not more than 1.0 ml. (taking body weight into consideration, equivalent to not more than 0.0004 ml. to a mouse). Therefore it is considered that incomplete adjuvant in such a dose would give no considerable damage to the liver or kidney of the injected person. In larger doses both Drakeol and Arlacel gave rise to liver damage. Drakeol had tendency to induce degeneration of liver cells. 0.6 ml. of Drakeol (a weekly dose of 0.1 ml. for 6 weeks) caused swelling of the liver cells partially accompanied with coagulation necrosis of the neighbouring liver cells. After a total dose of 1.8-2 ml. was administered, abnormal fatty tissue was observed over the surface of the liver and mesenterium, and liver cells were under vacuolar degeneration. Abnormal fat deposition was not demonstrated in any kind of cells by Sudan III staining. Arlacel A was prone to cause infiltration in Glisson's capsules, which appeared only after the sum total of 1.8-4.2 ml. was administered. Deposition of yellowish red colored substance was demonstrated in the interstitium of Glisson's capsules by Sudan III staining. No remarkable changes were observed in the liver cells. When Drakeol and Arlacel were given simultaneously in large amount, both degeneration of the liver cells and inflammatory changes in Glisson's capsules occurred at the same time. No pathological changes were induced in the kidney by either of two even after the largest dose was given. No mice died or suffered from arthritis during the course of this experiment.

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In patients of nasal allergy, the positive intracutaneous reactions ot several kinds of allergen extracts are often observed. For example, the positive reactions to ragweed pollen extract were obtained in the same percentage as those to the extract of Japanese cedar pollen. It seems strange that the positive reactions are obtained inspite of no ragweed colony in the district of Nikko. Therefore the authors consider that there is some problem on the present standard of the examination. From the results of examination on patients of nasal allergy in Tokyo and Japanese cedar pollinosis in Nikko, it seemed reasonable that the diagnostic borderline of intracutaneous reactions of ragweed pollinosis be set around 50 mm in erythema and 14 mm in wheal. While, in order to solve this problem using the present standard of the examination, the soulution of allergen extracts with high sensitizing power as ragweed pollen must be diluted. In conclusion, the authors propose that the dilution of allergen extract must be 1:500,000 or 1:1,000,000 in ragweed pollen.

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