We investigated beneficial effects of AIT with anticancer agents on survival of subcuta-neous tumor-bearing mice and suppression of artificial lung metastasis, and optimal schedule of administration of each treatment in vivo. 7-8 weeks old C 57 BL/6 mice were inoculated s.c. with 5×10⁶ B 16 melanoma cells, or ix. with 2×10⁵ B 16-F 10 melanoma cells. Mouse splenocytes were cultured with 3.5×10³ JRU/ml interleukin 2 for 14 days, and induced LAK cells were harvested. Anticancer agents (Cx), CDDP or MMC were given i.p. in mice. 1×10⁷ or 5×10⁷ LAK cells were given either s.c. around the tumor or i.v. respectively, and 1.4×10⁵ JRU/mouse IL-2 was administered s.c. for 6 days after LAK cell injection. Therapy groups were as follows #1: Cx day 3, AIT day 3-8. #2: Cx day 3, AIT day 6-11, #3: Cx day 8, AIT day 3-8. In therapy groups #1 and #2, we observed additive effects of AIT and anti-cancer agents in life-prolongation and suppression of lung metastasis. It was also shown that LAK induction in vivo was augumented by anticancer agents in groups #1 and #2, which might represent one of the mechanisms behind observed additive effects. Furthermore, our results suggest that therapeutic effects of the combination of AIT and anticancer agents depend on the schedule of administration. (J Nippon Med Sch 1997; 64 : 211-219)

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Collagen-induced arthritis (CIA) is useful animal model for human rheumatoid arthritis. We investigated the inhibitory effects of portal venous (p.v.) injection of type II collagen (CII) in CIA. The arthritis was suppressed by p.v. injection of CII before immunization for CIA induction. The p.v. route was more effective than intravenous or intragastric routes in the induction of tolerance in CIA. The dose of CII necessary for CIA suppression was 10 ug/20 g body weight in p.v. injection. Both anti-CII IgG and anti-CII IgG 2 a levels in serum were reduced in mice injected CII before induction of CIA. However, anti-CII IgG 1 levels did not differ between mice injected with CII and mice injected with buffer alone. Thus, the specific reduction in anti-CII IgG 2 a levels in mice treated by p.v. injection before immunization suggests that the suppression of CIA could be responsible for hypofunction of Th 1 cells. Reduction of anti-CII IgG and suppression of arthritis were observed when CII was injected through portal vein after immunization for CIA as well. (J Nippon Med Sch 1997; 64 : 220-224)

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Passive cutaneous anaphylaxis (PCA) reaction was developed by Ovary et al. as an animal model of mainly human type I allergic inflammation reaction. This is the most sensitive reaction for the detection of cutaneously snesitizing antibodies and provides a very effective means by which to investigate immunological reactions concerning the mechanisms of development and inhibition of allergic reactions, levels and specificity of antibodies, and the structure of antigens. These cutaneous inflammations mimic atopic dermatitis. Food ingestion has been pointed out as one of the worsend factors of atopic dermatitis. However, the body is generally protected against the invasion of high molecular substances such as non-ingested food by several barriers including digestive enzymes that break down complex food molecules into simpler substances and gastrointestinal mucosae. Accordingly, oral ingestion of food antigen seems to be physiologically and immunologically in conflict with the occurrence of dermatitis.
With a view to determining whether allergic dermatitis occurs after oral ingestion of food, the present study was carried out on animals, utilizing the PCA reaction. C 57 BL/6 Ncr j mice were immunized with immunogen derived from commercially obtained eggs. Wistar rats were used as a model of PCA reaction. The results of the present investigation are summarized as follows.
1) Blue spots of 10 mm diameter were observed as a PCA reaction 50 min or more after oral administration of antigen, and the blue spots reached maximum size (20-21 mm, 1.8-1.7 μg) after 90-120 min. The PCA reaction was induced 20 min or more after intravenous administration of antigen. When the maximum reaction was compared between the oral and the intravenous routes after 50 min (29.5 μg) and 90 min (1.8 μg) respectively, there was about a 16-fold difference in the capacity to induce inflammations.
2) The maximum PCA reaction values were 100 and 1, 600 for the oral and intravenous routes, respectively, there being a 16-fold difference between the two routes.
3) The minimum antigen concentration required to induce the PCA reaction was 10 mg/ml for the oral route and 0.01 mg/ml for the intravenous, there being a 1, 000-fold difference between the two routes.
4) Reactions with anti-egg mixture antibody and main egg constituents were specific. The PCA inhibition test results confirmed that the undigested structural portion of antigen was associated with the induction of PCA.
The present investigation demonstrated that there existed a mechanism by which type I allergy is induced via the gastro-intestinal tract. This fact indicates that part of the food ingested undergoes no change in its molecular structure when transferred to the blood, thus acting as a PCA inducing antigen. This phenomenon suggests that this animal model of human type I allergic dermatitis is a useful system that strongly suggests the association of food antigen with the development of allergic dermatitis. (J Nippon Med Sch 1997; 64: 225-231)

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We studied the clinical manifestations and causative agents in cases with allergic contact dermatitis due to eye drops who consulted the Department of Dermatology, Nippon Medical School during the peirod of 9 years between January, 1987 and December, 1995.
Among 66, 165 cases who visited the department during the studied period, 3, 903 were suspected as contact dermatitis or drug eruption and underwent patch test. 141 of 3, 903 cases (3.6%) were patch tested with eye drops and 49 cases (34.8%) reacted positively and were diagnosed as allergic contact dermatitis. Allergic ingredients were tested in 36 cases by the patch test with every ingredient.
The 49 cases diagnosed as allergic contact dermatitis by eye drops, included 17 males and 32 females. 81.6% of the patients were aged over 40 years. The most frequent ages were between 60 and 69 years. The period from onset to the first visit to us was less than two weeks in 54. 1% of the cases, but 3 cases consulted us after more than six months.
The determined allergens are noted in order of frequency as follows: 1) fradiomycin sulfate in 14 cases, 2) ketotifen fumarate in 6 cases, 3) dibekacin sulfate in 3 cases, 4) befunolol hydrochloride in 3 cases, 5) phenylephrine hydrochloride in 3 cases and 6) benzalkonium chloride, the preservative of eye drops in 3 cases. Fradiomycin sulfate showed cross reactions to other aminoglycosides in the present cases.
Every ingredient should be tested as much as possible, because there is a possibility that a preservative or a vehicle ingredient may be the allergen, though the incidence is low. (J Nippon Med Sch 1997; 64 : 232-237)

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The effect of anticancer agents, dacarbazine [DTIC (dimethyl-triazeno-imidazole-carboxamide)] and vincristine sulfate (VCR) on two human melanoma cell lines (G 361 and MeWo) was investigated. First we estimated the growth curve of two cell lines by using an isotope labelling technique with DNA, RNA and protein precursors to find the most appropriate conditions, and then, DTIC and VCR were added at various concentrations. When the drug was added alone, DNA, RNA and protein synthesis were suppressed in a dose-dependent fashion and the pattern of responses was similar between the two. When the two agents were administered in combination, responses were variable depending on the combinations of the concentrations of these agents. It was also observed that the response on DNA synthesis is not similar to that on RNA or protein synthesis, and that unbalanced growth can heppen when two anticancer agents, whose mechanisms of action against tumour cells, are different, are administered at the same time. These observations also differed between the two cell lines. It suggests that these diverse responses of melanoma cells against chemotherapeutic agents hinder the establishment of a sensitive combination chemotherapeutic regimen. (J Nippon Med Sch 1997 ; 64: 238-248)

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Hapten specific lymphocyte proliferative assay was used to measure systemic immunization induced by epicutaneously applied hapten, instead of the assay measured by ear swelling or footpad swelling. DNFB was painted on the shaved back of C 57 BL/6 on two consecutive days and 4 days later. DNFB sensitized lymph node cells (LNC) were obtained from the regional lymph nodes. Spleen cells were pulsed with an antigen after irradiation and used as antigen presenting cells (APC). Various numbers of LNC were cultured with various number of APC to determine the highest lymphocyte proliferation. This was observed when 2×10⁵ cells/well APC were cultured 4×10⁵ cells/well LNC.
In next step, this assay was used to investigate whether contact dermatitis is a required subject or not to sensitize. FB could not induce contact dermatitis on the painted region not only clinically but also histologically. FB, however, was able to induce hapten specific lymphocyte proliferation. This fact demonstrates that contact dermatitis is not always necessary in sensitization through epicutaneous application of hapten. (J Nippon Med Sch 1997 ; 64 : 249-255)

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It was demonstrated that the reaction of some cell species to peanut agglutinin (PNA) involved the maturity of the cells. We investigated the selective adhesion of circulating hemopoietic cells and lymphoid cells to hepatic sinusoidal cells. We experimented with thymocyte (TYC) models that could be easily isolated into PNA positive [PNA(+)] and PNA negative [PNA(-)] subsets, and transfused these subsets, labeled with 51Cr into the isogenic mice, then measured the radioactivity in the kidney, lung, spleen, liver, Peyer's patch (P.P.), and thymus, after 48 hrs. The radioactivity (% total dose recovered) in the liver and spleen represented the degree of cellular distribution and indicated that a significantly higher dose had accumulated in the liver and spleen than in other organs. Additionally, TYC-PNA(+), recognized to be immature cells, demonstrated high aggregation in the liver, while TYCPNA(-), recognized to be mature cells, demonstrated high aggregation in the spleen. Similar results were observed in our subsequent experiment using wheat germ agglutinin (WGA), other indicator of maturity, and spleen cells (SPC). Next, we transfused TYC-PNA(+) to mice at various intervals after 70% hepatectomy. The degree of aggregation (radioactivity/ weight) in the liver was reduced 1 day after hepatectomy, and that in the spleen increased proportionately. Whereas the aggregation degree of, TYC-PNA(-) in the same experiment did not change significantly.
We postulated that hepatic sinusoidal cells recognize and, sequentially, trap specific carbohydrates on lymphoid cells by a lectin, PNA or WGA, like receptor. Reviewing our previous report, adhesion to Kupffer cells indused SPC activation and proliferation, this recognition and trapping mechanism seems to play an important role of intrahepatic hematolymphoid system. (J Nippon Med Sch 1997; 64: 256-263)

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