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Biomedical subjects

N Okada

Publications and source records attributed to N Okada.

At least 415 records · Page 23Linked to original sources

Enterohepatitis in Mongolian gerbils (Meriones unguiculatus) inoculated perorally with Tyzzer's organism (Bacillus piliformis).

Enterohepatitis was produced in Mongolian gerbils by intragastric inoculation with Tyzzer's organism from natural infection of a gerbil. Death occurred in 50 to 60% animals 5 to 7 days postinoculation (p.i.). On day 3 p.i., when a few necrotic foci appeared in the liver, a large amount of bacterial antigen was present within ileocecal enterocytes and reticuloendothelial cells of the Peyer's patches. Neutrophil and monocyte infiltration was found in the lamina propria. On day 5 or 6 p.i. there was severe necrotizing and hemorrhagic ileotyphlocolitis. Bacterial antigen was abundant within not only enterocytes, but also smooth muscle cells of the ileum and jejunum as well as reticular cells of the mesenteric lymph nodes. On day 7 p.i. the intestinal lesions subsided in the presence of fewer bacteria, while necrotizing hepatitis became well developed. The results indicated that Mongolian gerbils were highly susceptible to the oral route of infection with the Tyzzer's organism.

Animals↗

Mutant monoclonal antibodies with select alteration in complement activation ability. Impact on immune complex functions in vivo.

Mutagenesis of mAb is a useful means for studying the biologic and pathologic functions of immune complexes. Treatment of the Hy-1.2 hybridoma-producing IgG2a-anti-TNP antibodies with ethylmethanesulfonate provided us with a mutant clone, producing antibodies with reduced capacity for C activation. The antibodies retained normal Ag-binding capacity, staphylococcal protein A reactivity, and association to FcR for IgG on murine macrophages. No significant polypeptide deletion or class-switch was observed, but a significant change in clonotype was revealed by IEF. Intravenous injection of the mutant antibodies in immune complex form induced different tissue distributions of Ag in mice; i.e., more in kidneys and less in spleen, and developed more mesangial deposits in renal glomeruli compared with those of the wild type. Moreover, the production of granulomatous lesions in vivo caused by immune complexes of TNP-Sepharose was augmented by using mutant antibodies. These lesions demonstrated an enhanced accumulation of macrophages with multinucleated giant cells. Availability of this kind of mutant mAb is thus helpful in the elucidation of the biologic functions and consequences of immune complexes.

Animals↗

Binding of C3 molecules to membranes via the SH-residue generated by the cleavage of a thioester bond can initiate complement activation.

Dithiopyridine (DTP)-dipalmitoylphosphatidylethanolamine (DTP-DPPE) was incorporated into liposome membranes to prepare DTP-liposomes. The DTP-liposomes could be lysed by reaction with the alternative complement pathway of any kind of serum tested. Activation of the alternative complement pathway has been shown to be mediated by the binding of C3 molecules to DTP on the liposomes via the SH-residue generated by the cleavage of thioester bond in the alpha-chain of the molecules.

Animals↗

SSA/Ro antigen expression in simian virus 40-transformed human keratinocytes.

SSA/Ro antigen is a soluble cellular component to which antibodies are frequently produced in patients with Sjögren's syndrome and systemic lupus erythematosus. Its exact location within the cell has yet to be determined. In this study we report the expression of SSA/Ro antigen in simian virus 40 (SV40)-transformed keratinocytes. The locations of SSA/Ro, U1RNP, and DNA antigens were studied by indirect immunofluorescence using monospecific antibodies. SSA/Ro antigen was detected in both the nucleus and cytoplasm of SV40-transformed keratinocytes tested with three monospecific sera. Primary cultured keratinocytes derived from adult human skin showed localized immunofluorescent staining within the nucleus. When Ca++ concentration of the medium was switched to 0.05 mM, these cells expressed cytoplasmic SSA/Ro antigens within 48 h. Depletion of the antibody activity with insolubilized human spleen extract abolished the staining. Surface expression of this antigen could not be detected in either primary or transformed cells. Localization of U1RNP and DNA was not altered. These results indicate that expression of SSA/Ro antigen in human keratinocytes is modulated by SV40 infection and that this antigen is expressed to a greater degree in cells that are less differentiated, transformed, or proliferating.

Autoantigens↗

Expression of CR2 (C3d receptor) on the cell membranes of adult T cell leukemia.

MT-2 cells, which produce human T-cell lymphotropic virus type I (HTLV-I), are known to have a complement receptor. We have established that the complement receptor is CR2 which binds C3d on immune complexes but not CR1. CR2 was also detected on ATL-3I cells but no complement receptor was detected on ATL-1K cells which lack ATL antigen (ATLA). Since CR2 is not detectable on normal T lymphocytes, the presence of CR2 on some ATL cells might suggest that ATL cells were derived from a particular minor lineage of T cells, or HTLV-I has a capacity to induce CR2, which has been demonstrated to be an a-type growth factor for B lymphocytes and to be a receptor for Epstein-Barr virus.

Antigens, Differentiation, B-Lymphocyte↗

Virulence-associated genetic regions comprising 31 kilobases of the 230-kilobase plasmid in Shigella flexneri 2a.

By random transposon Tn5 insertions, we previously identified six virulence-associated SalI fragments, B, D, F, G, H, and P, in the 230-kilobase plasmid pMYSH6000 of Shigella flexneri 2a. In this study, we analyzed the sites of 134 independent Tn5 insertions on four contiguous SalI fragments, B, P, H, and D, of pMYSH6000 and identified five virulence-associated regions; four were associated with inducing a positive Sereny test (Ser), invasion into epithelial cells (Inv), binding to Congo red (Pcr), and inhibition of bacterial growth (Igr), and one was associated with the Ser and Inv but not with the Pcr or Igr phenotypes. Hybridization studies revealed that these virulence-associated DNA regions were highly conserved among 15 other virulence plasmids of four species of Shigella and enteroinvasive Escherichia coli. These data indicate that at least seven separate genetic determinants on the virulence plasmid are required for full expression of the virulence phenotype of shigellae.

DNA Restriction Enzymes↗

Molecular genetic approaches to the pathogenesis of bacillary dysentery.

Bacillary dysentery is an invasive infectious disease of the human colon. At least three genetic loci on the chromosome and a huge plasmid have been implicated in its pathogenesis. Results obtained from molecular genetic studies, mainly of the genes, virG, virF and kcpA are discussed.

Animals↗

Fibronectin enhances respiratory burst of phagocytes stimulated by zymosan and immune complexes.

Plasma fibronectin (FN) has been demonstrated to serve as an opsonin involved in the ingestion of foreign particles by phagocytes. This study concerns the effect of FN exposure on the respiratory burst of normal human peripheral phagocytes, using a luminol-dependent chemiluminescence (CL) assay for measurement of reactive oxygen metabolites generated. FN enhanced, in a dose-dependent manner, the CL response of circulating monocytes stimulated, probably via beta-glucan receptor, with unopsonized zymosan. FN also increased the CL response of phagocytes to fresh serum-opsonized zymosan. When we used a glycolipid (ceramide pentasaccharide, CPS) incorporated on liposome membranes as an antigen, the immune complexes prepared between CPS and human IgG (as antibody) did not induce a CL response, differing from previous reports. Addition of FN to the immune complexes significantly enhanced the CL response of phagocytes. The role of FN in host defence is discussed.

Antigen-Antibody Complex↗

Several aspects of total genomic DNA transcription in a HeLa cell extract.

Several aspects of total genomic DNA transcription in a HeLa cell extract were described. (1) By using in vitro transcripts from total genomic DNA as probes, we elucidated several structures of short interspersed elements which are transcribed by RNA polymerase III (designated as Pol III/SINEs. (2) To know whether the repetitive sequence we cloned and sequenced comprises a major or minor family, we developed in vitro run-off transcription assay of total genomic DNA. (3) Unlike in vitro transcription of total genomic DNA from vertebrates, major in vitro transcripts from macronuclear total DNA of Tetrahymena were found to be tRNA themselves.

Animals↗