Biomedical subjects
Publications and source records attributed to K Onoue.
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We developed an efficient method for production of experimental polyarthritis. Native collagen (type II) from human costal cartilage was emulsified with incomplete Freund's adjuvant (IFA) containing the synthetic adjuvant N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP), and injected into one hind footpad of the PVG/c rat. This method reproducibly induced severe arthritis with high incidence, whereas the collagen alone in IFA produced mild arthritis with low incidence. MDP alone in IFA was shown to be minimally arthritogenic under identical conditions.
Water-soluble glycopeptides isolated from Lactobacillus plantarum and Staphylococcus epidermidis cell walls elicited a delayed type hypersensitivity (DTH)-like skin reaction in rats previously immunized with Mycobacterium tuberculosis cell walls, but not in unimmunized rats. Histological examination of the skin reaction sites in immunized animals revealed a close similarity of this skin reaction to a typical DTH reacton with respect to the time course of development and the types of cells that infiltrated into the skin reaction sites, which were characterized by a predominant infiltration of mononuclear cells at 48 hr. This DTH-like reaction was also demonstrated by immunizing the rats with the cell wall peptidoglycans of L. plantarum or S. epidermidis and skin testing them with homologous as well as heterologous peptidoglycans. The DTH-like reaction appeared to be caused by peptidoglycans that exist in common in the cell walls of phylogenetically distant bacterial species. Furthermore, it was also suggested that the putative antigenic determinants(s) might include both the glycan chain and part of the peptide moieties of the cell wall peptidoglycan rather than either of the single moieties.
Treatment of macrophages with cytochalasin E in combination with a lectin was found to stimulate the generation of superoxide anions (O2-) very efficiently. The macrophages stimulated with concanavalin A, phytohemagglutinin or wheat germ agglutinin released superoxide, but cells pretreated with cytochalasin E released much greater amounts of superoxide, without notable lag time, upon stimulation with the lectin. Wheat germ agglutinin was found to be the most efficient stimulant among the lectins tested. Superoxide generation in guinea pig macrophages was shown to be dependent largely on cytoplasmic glucose metabolism and to some extent on mitochondrial respiration, since the superoxide release was largely but not totally inhibited by 2-deoxyglucose and to a lesser extent by antimycin A or KCN. The method presented is sensitive and allows rapid assay of the superoxide-generating activity with only 1--5 X 10(5) macrophages for a single determination. In application of this technique, elevation of the superoxide-generating activity was shown with macrophages elicited by chemical inflammation or those obtained from mice after treatment with tubercle bacilli.
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Serum antibodies to acetylcholine receptors (AChR) in myasthenia gravis were surveyed by radioimmunoassay, using 125I-alpha-bungarotoxin-complexed receptors of human muscles and denervated rat muscles. Antibodies were detected more frequently with human AChR than with rat AChR. Furthermore, antibody titers to human AChR were not always parallel to titers observed in the rat receptors, indicating the heterogeneity of antibody specificities in different individuals. Correlation between antibody titers and clinical severity was better when the antibody concentration was determined with human AChR. About 90% of the myasthenic sera contained antibodies to AChR, and 40% also contained other antibodies such as thyroid autoantibodies. These findings suggest that myasthenia gravis is a multiple immunopathy.
For the detection and characterization of circulating immune complexes (CIC) in various liver diseases, a Clq binding test was used. Though the CIC level was almost normal in HB surface antigen (HBsAg) positive asymptomatic carriers, the level increased in patients with liver diseases. During acute exacerbation of chronic viral hepatitis, the CIC level reached peaks 1 to 3 weeks before and after the hepatic cell necrosis. Study of the sedimentation rates of CIC in various liver diseases showed CIC in the 19s-22s region and in the 7s-19s region. In acid buffer, CIC was dissociated into 5 to 6 components by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In one case of HBsAg positive severe chronic aggressive hepatitis, CIC was composed of HBsAg, IgG and another three or four undetermined components. During acute exacerbation of chronic hepatitis, minor changes of these dissociation patterns of CIC were observed.
Immune complexes in liver specimens from 10 patients with chronic liver diseases [2 with chronic persistent hepatitis (CPH), 3 with chronic aggressive hepatitis (CAH) of moderate activity, 3 with CAH of severe activity, and 2 with liver cirrhosis] were examined by a technique of direct immunofluorescence using FITC-labelled human purified Clq (FITC-Clq). FITC-Clq bound to the nuclei of all cells in liver tissue. After DNase treatment, positive nuclei were absent, but positive staining with FITC-Clq remained in amorphous deposits and hepatic cell membranes in the areas of piecemeal necrosis of four CAH patients. Since FITC-Clq could not be demonstrated in the liver tissue of CPH and liver cirrhosis which contained no piecemeal necrosis, positive fluorescence in the liver of CAH patients was thought to indicate immune complexes bound to FITC-Clq. The fact that these positive substances, however, were few in number, may be the result of physiological mechanisms of immune clearance which rapidly eliminate immune complexes from the body.
Fragmented sheep red blood cell (SRBC) membrane was used for detection of T-cells in liver biopsy specimens from patients with chronic active hepatitis. SRBC was separated with Lymphoprep, sonicated, then filtered through a 3 mu Millipore-membrane as a fragmented SRBC reagent. Tissue T-cells were stained by an indirect immunofluorescent technique using SRBC reagent and fluorescein isothiocyanate (FITC)-labelled rabbit anti-SRBC. Positively staining lymphocytes were present in portal tracts and in areas of piecemeal necrosis. There also seemed to be a positive correlation between the number of positively staining lymphocytes and the activity of chronic hepatitis; numerous lymphocytes being stained in areas of severe piecemeal necrosis. Our findings suggest that the fragmented SRBC technique for detection of T-cells is reliable and reproducible, that it could be used as a clinical routine method, and that it is useful for further elucidating the nature of host immune reactions on tissue levels.
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Follow-up studies were conducted on chronic liver disease patients treated 10 years previously with corticosteroid (CS), Hbs antigen (HbsAg) was measured in previously collected paraffin embedded liver sections by enzyme-labelled antibody technique. Of 57 cases examined, 38 cases were treated with CS or immunosuppressive agents and 19 cases were not treated with CS (control group). -- Two deaths occurred in the CS-treated group and 4 in the non-CS treated group. These patients were diagnosed as either the 2B type (severe activity) of chronic aggressive hepatitis (CAH) or as having liver cirrhosis. One death was found in the 10th year due to hepatocellular carcinoma in a chronic persistent hepatitis (CPH) patient with HBsAg diffusely distributed in liver tissue. No significant difference was found in the rehabilitation rate in HBsAg negative cases of the CS group versus the control group. In positive cases, the rehabilitation rate was found to be 66.7% in the CS group compared with 0% in the non-CS group.
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In the capillary tube migration system a synthetic muramyl dipeptide (MDP; N-acetylmuramyl-L-alanyl-D-isoglutamine), a part of bacterial cell wall peptidoglycans, inhibited the migration of peritoneal exudate macrophages from normal guinea pigs or rats. The migration inhibition was also caused by some MDP-containing peptidoglycan fragments from cell walls of Lactobacillus plantarum and Staphylococcus epidermidis. The migration inhibition could not be explained on the basis of macrophage migration inhibitory factor. A stereochemically highly specific structure of MDP required for its adjuvant activity was also required for the macrophage migration inhibition. These findings suggest that MDP and MDP-containing cell wall fragments may activate macrophages and that this activation may be important in the exertion of their adjuvant activity.
A study of 52 liver biopsies (47 hepatitis type B and 5 asymptomatic carriers) was performed to clarify the roles of HBe antigen (HBeAg), HB surface antigen (HBsAg) and HB core antigen (HBcAg). In this study, the Gudat classification was modified so as to classify the patterns of HB antigens into six reaction types including: type O (negative for both liver HBsAg and liver HBcAg), type III-A (characterized by a spotty HBsAg pattern) and type III-B (characterized from a sub-lobular to lobular HBsAg localization pattern). This classification enabled accurate prediction of the prognosis of hepatitis. Patients with positive serum HBeAg had either minimal hepatitis with mild clinical features or chronic aggressive hepatitis with severe clinical features. Ten patients negative for both HBeAg and HBeAb were all positive for liver HBcAg. In all 3 patients on corticosteroid administrations liver tissue was markedly positive for HBcAg and serum was usually positive for HBeAb.
The superoxide anion-generating capacity of human blood monocytes was measured by a sensitive and rapid method established by taking advantage of the fact that the generation of superoxide anions by monocytes was markedly enhanced by the combined stimulation of the cells with cytochalasin-E and wheat germ agglutinin. The activity was expressed by the initial rate of cytochrome c reduction after the addition of wheat germ agglutinin. The rate obtained with normal human monocytes was 0.73 +/- 0.19 nmol/min/10(5) monocytes (mean +/- SD, n = 10). Because of its sensitivity, the method required only 10(5) monocytes and can be used to follow the monocyte function in various patients. Preliminary data obtained with 15 cases of advanced cancer patients (0.29 +/- 0.10 nmol/min/10(5) monocytes) suggested a possible decrease of the superoxide-generating activity, at least in some state of cancer patients. It appears that measurement of superoxide-generating activity will be meaningful to monitor monocyte function.
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Histocompatibility-linked restriction of macrophage-T lymphocyte interaction in antigen-induced MIF production by sensitized lymphocytes was examined, by using combinations of inbred strain 2, strain 13, and JY-1 guinea pigs. The effective interaction of the antigen-bearing macrophages with the immune T lymphocytes was observed when the donor of the antigen-bearing macrophages and that of the immune lymphocytes shared Ia antigens of the major histocompatibility complex. Identities of B antigens and S antigens were not important for this cooperation. It was further demonstrated that the previously reported soluble factor derived from LPS-stimulated peritoneal adherent cells (macrophages) could help antigenic activation of the immune lymphocytes across the strain barrier provided a small number of macrophages (0.01%) from syngeneic strain were present. These results show that the presence of macrophages is absolutely required to present antigen to immune T lymphocytes in a genetically restricted manner and the soluble factor from macrophages appears to give a nonspecific effect on the lymphocyte activation in addition to or in collaboration with antigenic stimulation.