Acta Medica Okayama volume31 issue3

Further purification and characterization are reported on rat cytosol cornin (RLCC), an antimitotic substance. Fraction I (purified RLCC) was purified more than 10-fold from crude RLCC with Sephadex G-50 column chromatography and showed a remarkable inhibitory effect on division of inseminated sea urchin eggs and mouse fibroblast cells. Fraction I was observed as one spot, and the molecular weight was estimated to be about 25,000 by thin layer gel filtration. Fraction I contained protein (92%) and RNA (8%), but the antimitotic activity was scarcely affected by treatment by pancreatic RNase. The protein of Fraction I was separated into two bands by SDS-polyacrylamide gel electrophoresis, and the molecular weight was estimated as 10,000 and 15,000, respectively. The 50% inhibition dose of Fraction I on the first division of inseminated sea urchin eggs and on proliferation of mouse L cells was about 2.5 X 10(-5) g/ml and 5 X 10(-4) g/ml, respectively. The yield of fraction I was about 35 mg from 100 g rat liver.

The present study was conducted to investigate the usefulness of the direct leucocyte migration agarose method for studying cell-mediated immunity in vitro. Comparative studies of the purified protein derivative (PPD) skin test and the leucocyte migration inhibition test (LMIT) in which PPD was used as test antigen indicated a significant qualitative and a weak quantitative correlation between these two tests. Furthermore a positive correlation was found between the LMIT and the macrophage migration inhibition test (MIT) using ultrasonicated authochthonous tumor antigen. Comparative studies of the LMIT, MIT, PPD skin and DNCB tests on the same patients showed that cases responding positively to the the PPD skin and DNCB tests tended to respond positively to the LMIT and MIT. Patients with digestive organ cancers were examined by the LMIT. With the advance of cancer, decreased positive test test rates were found. After gastric cancer operations the LMIT findings were divided into two groups: one type changed from positive to negative, and the other type changed from negative to positive. The former response was suggestive of a successful operation, and the latter response was suggestive of a non-curative operation. These results indicated that the direct leucocyte migration inhibition agarose test was useful investigating cell-mediated immunity.

Twenty cases of suspected drug induced liver injury (16 cases of which satisfied the criteria for at least a query positive diagnosis as based on the Fourth Congress of "Drugs and the Liver" in Japan) were studied by the whole blood culture technique of lumphocyte blast transformation. The positive rate with this technique was 10%, and no more than 15% even with the addition of one query positive. One reason for the low positive rate was that there was not only an allergic mechanism at work in the study group but that liver injury due to direct cytotoxicity of the drug was involved also. For a drug such as chlorpromazine with strong cytotoxicity for lymphocytes, it was difficult to demonstrate a relationship between allergic mechanisms and the drug with this method.

1. Three cases of acute rejection were detected by macrophage migration inhibition tests (MIT) conducted directly on seven patients who had received renal allografts. The macrophage migration inhibitory factor (MIF) activity was positive in all cases 1-2 days before the appearance of acute rejection. 2. After the administration of a high dose of Solu-Medrol (1g/day for 3 days) to suppress the acute rejection, MIF activity recovered to its normal level 3 days later. These findings seem to indicate that MIT yields immunologically useful criteria for the early detection of an acute rejection.

Mice were trained in an avoidance learning task. The incorporation of 3H-leucine into the hippocampal regions of trained mice was higher than that of control mice. When mice were injected with cycloheximide, a strong inhibitor of protein synthesis, impairment was evident in acquisition of learning. Cycloheximide produced morphological changes in mitochondria and microtubules of some brain axons. It is suggested that the cycloheximide-induced learning impairment may be due to the blocking of the synthesis of the specific protein necessary of neutral conductivity.

The relationship between immune insulitis and glucose tolerance was investigated in three groups of mice following active immunization with different components of bovine pancreatic hormone. An abnormal blood glucose level was observed in the three groups ranging from 33.3% to 87.5% of sensitized mice. A relationship was not present between the glucose tolerance response and the presence of insulitis or anti-insulin antibody in the blood of sensitized mice. However, all sensitized mice with a marked decrease in glucose tolerance were found to have insulitis. In animals without established insulitis and with no demonstrable anti-insulin antibody, abnormal glucose tolerance was noted. This latter condition occurred more frequently with recrystallized insulin than with a-component and did not occur with monocomponent insulin. These findings seemed to indicate that two distinct processes involving some circulating antibodies with anti-insulin antibody and insulitis might be involved in the development of the observed glucose tolerance abnormality.