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HB surface antigen (HBs Ag) was detected using the enzyme-labelled antibody technique on routinely processed liver biopsy material fixed in Bouin's fixative and embedded in paraffin. Of 85 examined specimens, 45 cases were HBs Ag positive by both the immunofluorescent test and the enzyme labelled antibody technique. The remaining 40 cases were negative by both techniques. The specificity of HBs Ag detected by the enzyme-labelled antibody technique was confirmed by the blocking test using guinea pig specific HBs antibody. The results indicate that the enzyme-labelled antibody technique may be useful for detecting HBs Ag on routine paraffin sections.

Hepatitis B core antigen (HBc Ag) and hepatitis B surface antigen (HBs Ag) were detected in the liver tissue of a patient with chronic aggressive hepatitis by the immunofluorescent complement technique. The presence of anti-HBc was examined by the same method in 67 human sera previously tested for HBs Ag, anti-HBs and s-GPT levels. HBc Ag was localized mainly in the nucleus and sometimes in the cytoplasm of the hepatic cells. HBs Ag was found only in the cytoplasm. The focal area of HBc Ag positive hepatic cells seemed to correspond to the HBs Ag positive cells. Double staining demonstrated the simultaneous presence of HBs Ag and HBc Ag in individual cells. Anti-HBc positive serum was found in 46 (68.7%) cases. Forty-eight (71.6%) indicated a combination of HBs Ag and anti-HBc.

In order to approach human cancer immunotherapy, the author carried out the immunotherapy with BCG on mice having homotransplanted cancer, observed the posttransplantation results with lapse of time, conduced daily macrophage inhibition test (MI test) and found the immunotherapy to be effective. At the same time the MI test proved to be a useful criterion in determining the course of cancer progress and effectiveness of the immunotherapy.

Phosphate-binding protein(s) was found in the inner mitochondrial membrane of calf heart by Sephadex G-200 and G-25 gel filtration. The binding activity was inhibited by N-ethylmaleimide and competed by a large amount of cold phosphate. The amount of phosphate bound to the fraction was 29 nmoles per mg of protein. Affinity chromatography with phosphate-bound Sepharose 4B confirmed the presence of phosphate-binding protein(s) in the active fraction of mitochondrial membrane fractionated by gel filtration.

In order to formulate an early diagnostic method for acute rejection after kidney transplantation, macrophage migration inhibition test (MIT) was carried out with lapse of time after inbred rat kidney allotransplantation. The mean survival time of rat kidney allograft was found to be 7.07 +/- 1.34 days. On the other hand, in the group treated with rabbit anti-rat lymphocyte serum (ALS) the mean survival time was lengthened to 14.15 +/- 2.14 days (p less than 0.05). The corresponding antigen used for MIT was prepared with donor kidney by ultrasonication, and its protein concentration at 180 mug/ml was the most optimal as not to elicit non-specific inhibition of macrophages. In the control group, activity of macrophage inhibitory factor (MIF activity) turned positive 3 days after the transplantation, and it became strongly positive by 5 or 7 dyas at the period when rejection crisis appeared frequently. ALS-treated group showed a lower MIF activity than the control group (p less than 0.05) and on 7-12 dyas before rejection crisis appeared frequently, MIF activity became strongly positive. These findings suggest that this MIT is simple and will be proved to be useful in predicting the acute rejection as well as in controlling the immunosuppression.

Human adenovirus type 12 (Ad 12) was inoculated through subtentorial route into inbred newborn mice (C3H/BifB/Ki), and sequential changes of the brain and tumor induction were examined by histological and immunofluorescent methods. Two days after virus inoculation, Ad 12 specific tumor antigen (fluorescent T-antigen) appeared in the cells of ependymal and subventricular matrix layers, choroid plexuses and leptomeninges in the subtentorial as well as the supratentorial brains. After 10 days, these fluorescent positive cells decreased gradually in number but still remained focally beneath the ependyma. Sixty days later, early tumor nodules were detected in the same regions in which remained the fluorescent cells. After 107 days, neurological signs and well-developed tumors were noted in 25 of 63 (30.1%) mice examined. In the cerebellum, both of T-antigens and tumors were limited around the IVth ventricle, but not in the granular layers. Histomorphologically, the tumors were of primitive neuroectodermal origin and consisted of the cells resembling immature matrix cells in the subventricular zone. These findings strongly suggest that the virus has a selective affinity to the remaining matrix cells, but not to cerebellar granular cells, at least, in newborn mice.

Heterokaryon formation and Rous sarcoma virus (RSV)-induction were studied by fusion of RSV-transformed human embryonic cells with chick embryo fibroblasts in the presence of lysolecithin. Heterokaryon formation was observed by autoradiography. RSV-induction was identified by focus formation, electron microscopy and density gradient centrifugation of 3H-uridine-labeled particles. The most effective concentration of lysolecithin for virus induction was 10 mug/10(6) cells/0.1 ml. Efficiency of lysolecithin in virus induction was not less than that of ultraviolet-inactivated Sendai virus (UV-HVJ).

A cell line (HGC-27) was established by culture of the metastatic lymph node from a gastric cancer patient diagnosed histologically as undifferentiated carcinoma. HGC-27 cells were polygonal or short spindle-shaped and adhered to glass surfaces as a monolayer. The cells were probably derived from gastric cancer cells, as their origin from mesenchymal tissues can be excluded morphologically and enzyme-histochemically. Enzyme activities were generally negative or low, except for adenosine triphosphatase, lactic dehydrogenase and leucine aminopeptidase. These scanty findings might reflect the undifferentiated character of the original tumor cells. The cloning efficiency was 5.3% in liquid medium and 1.0% in soft agar. The doubling time was about 17 hr. Chromosomal analysis revealed a mode of 109 and 110 chromosomes.

A case with prolonged bacterial infection accompanied by an abnormal serum protein which migrated in the post-gamma region on electrophoresis is presented. The abnormal protein was identified as IgG with gamma-type light chain moiety. The patient suffered from prolonged pneumonia and cholecystitis, Bone marrow aspiration and skeletal x-rays did not indicate multiple myeloma.

Es wurde Untersuchungen an Mausen mit dem Rinder-Monokomponente-Insulin und der Rinder-a-Komponente durchgefGhrt, urn den Nachweis zu erbringen, ob das Monokomponente. Insulin oder die a-Komponente als ein Insulitis-erzeugendes Antigen dienen kann. Dabei wurden die Tiere mit den Substanzen, die jeweils mit Freund's complete adjuvant wiederholt verabreicht wurden, aktiv immunisiert und weiterhin untersucht auf eine dadurch bewirkte Insulitis. (1) Bei den mit dem Rinder-MonokomponenteInsulin sensibilisierten Gruppen kam die Insulitis bei 1 von 8 Fallen in der 20. Woche nach der ersten Sensibilisierung und bei 5 von 10 Fallen in der 28. Woche zur Erscheinung. (2) Bei den mit der a-Komponente behandelten Gruppen liet3 sich die Insulitis bei 0 von 9 Fallen in der 20. Woche nach dem Sensibilisierungsbeginn und auch bei 1 von 10 Fallen in der 28. Woche nachweisen. Diese Ergebnisse zeigen, dat3 das Monokomponente-Insulin als ein Insulitis-erzeugendes Antigen wirken kann. Dagegen war nur ein Fall von Insulitis befallen unter 19 Tieren, die mit der a-Komponente behandelt wurden.

The surface structure of myeloma cells was examined by scanning electron microscopy. The cells were collected from the pleural effusion of a multiple myeloma patient and purified by Conray-Ficoll gradient sedimentation. The cell size ranged from 8 mu to 12 mu in diameter and the microvilli were from 0.8 mu to 1.2 mu in length. The surfaces of the majority of the observed myeloma cells were more villous than lymphocytes.

An anomalous zymogram of lactate dehydrogenase (LDH) in the serum from a patient with liver cirrhosis was reported. Agar-gel electrophoresis of serum showed an extra LDH band close to the anodic side of LDH5 and a wide band of LDH5. Gel filtration of patient's serum in Sephadex G-200 demonstrated an abnormal LDH fraction eluted between immunoglobulin G (IgG) and macroglobulin in addition to a normal LDH component. Chromatographically abnormal LDH was demonstrated on agar gel as extra and wide LDH5 bands and resembled closely human hepatic LDH in various physico-chemical properties such as inhibition by urea or substrate, stability against heat, and Michaelis-Menten's constant. Immunological analyses demonstrated that abnormal LDH could be in the state combined with IgG. Molecular weight of the complex estimated by gel filtration was approximately 300,000. Mixtures of the heated patient's serum with normal or patient's hepatic LDH showed abnormal LDH fraction by gel filtration, whereas abnormal fraction was not demonstrated when heated normal serum was mixed with normal or the patient's hepatic LDH. These results strongly suggest that the occurrence of anomalous LDH zymogram in patient's serum is due to a formation of LDH-IgG complex, which is based on the binding of essentially normal hepatic LDH and abnormal IgG.

Experimentelle Produktion der Immun-Insu1itis wurde aufgrund der aktiven Immunisierung der Mause vom dd-Stamm durch wiederholte Gabe vom rekristallisierten Rinderinsulin im Abstand von 4 Wochen unternommen. Wahrend der Zeitdauer vom 3. Tag bis zur 28. Woche nach der ersten Sensibilisierung wurden serologische sowie histo1ogische Untersuchungen an diesen Tieren vorgenommen. Dabei ergaben sich fo1gende Befunde: (1) Die Immunlnsulitis kam bei allen von 58 Fallen bis zu 16 Wochen nach dem Sensibilisierungsbeginn nicht zur Erscheinung, und trat bei 2 von 8 Fallen erst in der 20. Woche und dann bei 3 von 8 Fallen in der 28. Woche in die Erscheinung. (2) Kein signifikanter Unterschied bestand in Hinsicht des insulinverbindenden Antikorpertiters im Blut zwischen den Fallen mit und ohne Immun-Insulitis in der 20. Woche sowie in der 28. Woche. (3) 1m Zeitlauf gab es aber eine gute Koinzidenz zwischen der Entstehung der Immun-Insulitis und der Herabsetzung des Antikorpertiters im Blut. (4) Untersuchungen des Pankreas mit Hi1fe der direkten Fluoreszenz-Antikorpermethode ergaben keine erkennbare spezifische Fluoreszenz innerhalb der Langerhansschen Inseln. Diese Untersuchungsergebnisse liefern der Ansicht einen Beweis, da~ die Insulitis, die fUr den mensch1ichen Diabetes mellitus spezifisch ist, mindestens zum Teil durch einen immuno1ogischen Mechanismus entstehen konnte.

The macrophage migration inhibition activity [MI activity) was stable in sensitized lymphocyte-to-marcophage ratios of 1:5 to 1:20 in mice. Antigen protein concentrations under 100 mug/ml did not induce nonspecific macrophage migration inhibition. Inhibition of tumor proliferation and survival was observed after a combined injection of BCG and MH-134 cells. After a single injection of MH-134 tumor cells, MI activity was reinforced and prolonged, demonstrating the clear effects of BCG as adjuvant. In DDS mice MI activity was weakened in the regional lymph node after a subcutaneous injection of just above or below 10(5) Ehrlich cancer cells previously treated with mitomycin C. This finding suggests the presence of an optimal tumor antigen concentration.

A case of alcaptonuria combined with aortic insufficiency was found in a 28-year-old male. The patient was palpitating at admission. The daily excretion of homogentisic acid was 2.0-6.0 g. Electrocardiography indicated atrial fibrillation and left ventricular hypertrophy with a ST-T change and right axis deviation. Cartilage tissues in the knee-joints showed no pigmentation. Vertebral X-ray revealed no calcification. The patient's history disclosed a family intermarriage in his grandparents. The patient's mother noticed the presence of black stains on diapers in his infancy and brown pigmentation on the skin and sclera in childhood. No kin had similar symptoms.

Neocarzinostain (NCS) was first used by Hiraki and his colleagues for induction chemotherapy in acute leukemia. This new anti-tumor agent is a polypeptide with a high molecular weight of 10,700 daltons. Anti-NCS antibody was produced in rabbits administered NCS intramuscularly with or without adjuvant. The production of anti-NCS antibody in patients treated with NCS was investigated. Forty three leukemia cases of various types were examined totally 65 times. Two mg of NCS for four consecutive days by intravenous drip infusion followed by 7 to 10 days of pause was repeatedly administered. The total amounts ranged 8 to 174 mg and the total periods 4 to 87 days. The methods used to measure the antibody titer are the passive hemagglutination (PHA) test on microplate and the passive cutaneous anaphylaxis (PCA) reaction in guinea pigs. The sera of all patients showed only non-specific agglutination at less than 2(3) dilution by PHA test, and to confirm these results four patient sera were tested by PCA reaction. The production of anti-NCS antibody was not detected in patients by PHA test and PCA reaction. The anaphylactic reaction and other adverse reactions due to anti-NCS adtibody production were not demonstrated in patients. Anti-NCS antibody was not detected by these experiments in the dose schedule administered.

"Smoldering acute leukemia", a variant of acute myelogenous leukemia, has been recognized with frequent incidence in recent years. This is chracterized by benign clinical course, poor physical findings, leukopenia and mild anemia in the peripheral blood, and apparent infiltration of abnormal myeloblasts in the bone marrow. Immunological studies of the host defence mechanism were made, because the pathogenesis of its "smoldering" course has never been well understood. Nine cases, seen during last 2 years, were investigated for immunological profile, especially the cellular immunity. Purified protein derivative (PPD) skin test, i.e., tuberculin test, was found to be positive in 8 of 9 cases (88.9%). Dinitrochlorobenzene (DNCB) sensitization test showed to be positive in 4 of 6 cases examined (66.9%). Peripheral lymphocyte balstogenesis by stimulating with phytohemagglutinin (PHA) was evaluated using the smear counting method. The blastoid lymphocyte ratio was 55% at the median value (range: 31-68%), compared with 63% in normal young control (age: 25-32) and 41% in normal aged control (age: 60-75). In this report, the cellular immunity in smoldering acute leukemia was proved to be preserved at the normal level and to be more competent than that in aged group. The preserved cellular immunity is considered to explain the phenomenon of "smoldering", in other words, the exacerbating proliferation of leukemic cells is suppressed by immuno-surveillance system.

1. When the various anticancer agents are injected intravenously to normal rabbits and intraperitoneally to normal mice, it seems that the serum properdin levels fall transitorily for some hours after administration with a small dose and then keep rising, but with a massive dose it continues to fall from the beginning. 2. The properdin level is decreased considerably by Thio-TEPA and Carzinophilin; moderately by Mitomycin C; and slightly by M. H. OX-substance hardly changes the level and 8-azaguanine rather has a tendency to raise the level. 3. The administration of most anticancer agents seems to suppress the properdin system. 4. The influence of these agents on human serum properdin is similar to that of rabbits. 5. The properdin levels keep at high titers in the group to which the agents act effectively on the cancer, but the levels fall down more rapidly and animals die earlier in the group to which the agents act ineffectively on the cancer as compared with the control group.

For the purpose to reveal whether or not the liver and the cell organellae are responsible for the abnormal metabolism of polysaccharides found in cancer bearing individuals, the author analyzed the liver and ascites with tumor cells of AH 130 hepatoma bearing rats biochemically with some histochemical observations. A quantitative increase in polysaccharides accompanied by the production of unusual polysaccharides is found in the supernatant of liver from cancer bearing rats, but not from mitochondrial or microsomal fractions. Tumor cells themselves and ascites fluid do not contain the abnormal polysaccharides found in the liver supernatant.

1. The properdin levels in sera from mice bearing Ehrlich ascitic carcinoma and from rabbits with Brown-Pearce carcinoma decrease inversely with the increase of the ascites or the tumors. In the incipient period of tumor transplantation, the level rather rises. When the tumor is proliferating or large, the level keeps falling or is low. On the contrary, when the tumor is regressing or disappears, the level elevates or reverts to that before transplantation. Strong A and R III mice with spontaneous mammary cancer have markedly low serum properdin levels as compared with those of healthy mice. 2. The properdin levels are less than 2 units per milliliter of the serum in 44.4 per cent of patients with gastric cancer, in 18.2 per cent of ones with non-malignant tumor and in 18.2 per cent of ones with gastric or duodenal ulcer. The abnormal low level has been found in 33.3 per cent of patients without recurrence, who had undergone extended radical gastrectomy combined with radical lymphadenectomy for gastric cancer. 3. Some correlation can be seen between the serum properdin levels and the degree of progress of gastric cancer. 4. The cancer patients with low total serum protein have lower serum properdin levels than those having nomal protein. 5. As for influence of surgical operation on the serum properdin levels, there is observed a tendency that a minor operation causes the levels to increase and a major operation causes the levels to fall. 6. It has been inferred that the properdin system could be one of the host natural resistance against cancer.