MICROBIOLOGY and IMMUNOLOGY
Online ISSN : 1348-0421
Print ISSN : 0385-5600
ISSN-L : 0385-5600
Volume 37, Issue 5
Displaying 1-13 of 13 articles from this issue
  • Osamu Nakagomi, Toyoko Nakagomi
    1993 Volume 37 Issue 5 Pages 337-348
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
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  • Mohamed A. El-Farrash, Takao Masuda, Marcelo J. Kuroda, Shinji Harada
    1993 Volume 37 Issue 5 Pages 349-357
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The effect of host cell factors on infectivity of human immunodeficiency virus type 1 (HIV-1) was studied by infecting a monoblastoid cell line (U937) or a T-cell line (MOLT-4) with a highly infective single clone of HIV-1 and comparing the infectivity of the produced viruses to different cell lines. Chronically infected U937 cells consistently produced viruses with minimal infectivity. This phenotypic change was host-dependent as the back-passage of the U937-produced low infective viruses into MOLT-4 cells resulted in regaining their original high infectivity. Southern and Northern blot analyses of the HIV-1 grown in U937 cells did not reveal any genomic difference between it and the virus grown it MOLT-4 cells. The radioimmunoprecipitation analysis of viral proteins showed that the HIV-1-infected U937 cells had a different pattern of envelope glycoproteins and core proteins, which well correlated with the low infectivity of the produced viruses. This experimental system using MOLT-4 and U937 cell lines would be useful to further explore host cell factor(s) which play an important role in the regulation of HIV-1 infectivity.
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  • Wei-June Chen, Su-Lih Chen, Ay-Huey Fang, Ming-Tsan Wang
    1993 Volume 37 Issue 5 Pages 359-363
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    A protein A-gold-silver (pAgs) staining was developed to detect dengue virus antigens in cultured cells. The method can be carried out in either newly-subcultured or monolayered cells. Dengue virus-inoculated C6/36 clone of Aedes albopictus cells and human endothelial cells appeared brown-yellowish color on the peripheral membrane of the infected cells. In many cases, the infected C6/36 cells appeared darker than that of the infected endothelial cells. The positive results from the inoculated C6/36 cells usually appeared as early as 2 days post-inoculation for types 1, 2, and 4 of dengue viruses and 3 days for the dengue 3 virus. The same batch of specimens detected by direct immunofluorescence antibody test (DFA) showed positive 4 days post-inoculation for the types 2, 3, and 4 of dengue viruses and 6 days for the dengue 1 virus. The result also showed that all pAgs-positive specimens were also DFA-positive, but not vice versa. It suggested that pAgs is not only sensitive but also specific for dengue virus detection from inoculated cultured cells.
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  • Yoichiro Kino, Yoichi Minamishima
    1993 Volume 37 Issue 5 Pages 365-368
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    A simple and effective method for the detection of antibodies to herpes simplex virus (HSV), human cytomegalovirus (HCMV) and varicella-zoster virus (VZV), has been established using the passive hemagglutination assay (PHA) in combination with viral specific glycoproteins. The results obtained with the PHA were compared with those from neutralization (NT) and complement fixation (CF) tests. The PHA test for each of the herpes viruses appears to compare favorably with the other assays tested. The specificity and sensitivity of HSV PHA to NT were 100%, whereas the specificity and sensitivity of HSV CF test to NT were 98% and 100%, respectively. For HCMV, the specificity and sensitivity of PHA to NT and PHA to CF were 100%. Similarly, the specificity and sensitivity of VZV PHA to NT were 100%. Because of the low sensitivity of the VZV CF, the sensitivity of CF to NT was 83%. Furthermore, the range of antibody titers and their absolute levels obtained in the PHAs were significantly greater than those in the NT and CF tests.
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  • Yoichi Moroi, Yasuhiro Koga, Kazuhiko Nakamura, Masumi Ohtsu, Genki Ki ...
    1993 Volume 37 Issue 5 Pages 369-381
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The role of lck gene in T cell proliferation and differentiation was investigated with transgenic mice carrying human lck cDNA whose expression was regulated by the promoter of mouse H-2Kb and the enhancer element of mouse IgH. RNase protection assay revealed that the lck transgene was expressed in the thymus and spleen, whereas immunoblot analysis demonstrated that amounts of p56lck in freshly isolated lymphoid organs were almost equal between transgenic mice and negative littermates. Cell-surface marker analyses of the thymocytes and peripheral lymphocytes revealed no remarkable difference between both groups. Notable finding is that the thymocytes from transgenic mice showed a significant proliferative response to the stimulation with IL-2, but not the thymocytes from negative littermates. Further analysis revealed that CD4+8- single positive thymocytes proliferated in response to IL-2. While surface expression levels of IL-2Rα and IL-2Rβ of these CD4+8- thymocytes from transgenic and control mice were almost equal before stimulation with IL-2, the expression of IL-2β3 was induced only in transgenic thymocytes after stimulation with IL-2. Immunoblot analysis demonstrated that the expression of p56lck of transgenic thymocytes was not down-reguated at 4hr after stimulaion with IL-2, whereas p56lck of control ones were not detectable any more at 4hr after stimulation with IL-2. Moreover, in vitro kinase assay substantiated such unchanged expression of p56lck in the thymocytes from transgenic mice: the kinase activities of p56lck did not decrease in thymocytes from transgenic mice after stimulation with IL-2, while kinase activities of control ones were significantly down-regulated by stimulation of IL-2. These results suggested that a significant proliferative response found in the thymocytes from lck-transgenic mice after the stimulation with IL-2 was caused by a constitutive expression of p56lck in these thymocytes even after the stimulation. Our findings, therefore, support a possibility that p56lck may play a role in the IL-2R-mediated signaling system in CD4+8- thymocytes.
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  • Didier Hober, Faiza Ajana, Michel Boniface, Rocio Estrada, Pierre Emma ...
    1993 Volume 37 Issue 5 Pages 383-390
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The level of sCD23 produced in the course of human immunodeficiency virus (HIV) infection was measured in patients grouped according to the Centers for Disease Control by using an immunoradiometric assay. Soluble CD23 was evaluated in supernatants of peripheral blood mononuclear cell (PBMC) (106cells/ml) stimulated by phytohemagglutinin (PHA). Compared with healthy controls (m±S.D.=1.0±0.34U/ml, n=7), higher values were observed in some of the patients of group II (asymptomatic) (m±S.D.=2±1.33, n=9) and some of the patients of group IV (AIDS) (m±S.D.=1.3±1.40, n=8). Those results prompted us to compare the plasma levels of sCD23 in group II and group IV HIV-infected patients and in healthy individuals. Soluble CD23 plasma levels in healthy patients (n=42) ranged from 0 to 1.5U/ml (m±S.D.=0.9±0.33), in group II patients (n=17) from 0 to 3U/ml (m±S.D.=0.92±0.83) and in group IV patients (n=73) from 0 to 2.9U/ml (m±S.D.=1.15±0.71). The differences between the patients and the healthy individuals were not statistically significant but individual sCD23 values higher than 2U/ml were obtained in 6% of the group II patients and 16.7% of the group IV patients. Increased values of sCD23 were obtained in plasma from patients with secondary infectious diseases (groups IV-C1 and IV-C2) and from patients without secondary infectious diseases (group II, group IV-A and group IV-B). Elevated values of sCD23 were detected even in patients with low counts of CD4+ T cells and CD8+ T cells in their peripheral blood. sCD23 has numerous activities including control of IgE synthesis and cytokine-like properties. Our results show a disarray of sCD23 in HIV-infected patients which could be involved in drug reactions, allergic manifestations and the IgE-level increase. Further investigations should attempt to define the role of sCD23 in clinical manifestations of HIV infection.
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  • Claudio Chiesa, Lucia Pacifico, Fulvio Nanni, Anna Maddalena Renzi, Gi ...
    1993 Volume 37 Issue 5 Pages 391-394
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    From 1981 to 1991, 37, 666 human, animal, food and environmental samples were cultured for Yersinia pseudotuberculosis using direct plating methods and/or cold enhancement techniques. Despite an intensive surveillance and adequate culture methods, Y. pseudotuberculosis was isolated from stools of 0.05% (5/9, 720) of patients with acute enteritis, and alimentary tracts of 0.1% (10/6, 849) of apparently healthy animals. No Y. pseudotuberculosis strains were recovered from stools of 4, 726 healthy controls nor from the appendices (656), mesenteric lymph nodes (84), and stools (421) of 656 patients operated for suspected appendicitis. Of the 10, 842 food and 4, 368 environmental samples, none yielded positive cultures for Y. pseudotuberculosis.
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  • Nobuhiro Fujii, Kouichi Kimura, Noriko Yokosawa, Keiji Oguma, Teruo Ya ...
    1993 Volume 37 Issue 5 Pages 395-398
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The complete nucleotide and deduced amino acid sequence of the nontoxic component of botulinum type E progenitor toxin is determined in recombinant plasmid pU9BUH containing about 6.0kb HindIII fragment obtained from chromosomal DNA of Clostridium butyricum strain BL6340. The open reading frame (ORF) of this nontoxic component gene is composed of 3, 486 nucleotide bases (1, 162 amino acid residues). The molecular weight calculated from deduced amino acid residues is estimated 13, 6810.1. The present study revealed that 33 nucleotide bases of 3, 486 are different in the nontoxic component gene between C.butyricum strain BL6340 and C. botulinum type E strain Mashike. This corresponds to the difference of 17 amino acid residues in these nontoxic component.
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  • Hiroshi Ogawara, Kei-ichi Kuma, Takashi Miyata
    1993 Volume 37 Issue 5 Pages 399-403
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    β-Lactamase is an enzyme which catalyzes the hydrolysis of the β-lactam ring of penicillins and cephalosporins. By similarity analysis of amino acid sequences in a database, the amino acid sequence deduced from the nucleotide sequence of the upstream region of cytochrome c oxidase subunit II from Paracoccus denitrificans was found to have an unusually high score of homology to that of a portion of β-lactamases from Gram-negative bacteria. Furthermore, the nucleotide sequences corresponding only to this region had a very high score of similarity among them. The phylogenetic tree constructed on the basis of the amino acid sequences was in accord with that constituted on the 5S rRNA's. Moreover, the molar G+C contents and the codon usage were similar to those in their respective bacteria. It is suggested, therefore, that the nucleotide sequence in P. denitrificans was positioned by a transfer of a part of a β-lactamase gene formed as a result of gene duplication or it was formed by a deletion of the essential region of the β-lactamase gene, although no β-lactamase gene has been yet detected in P. denitrificans.
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  • Sumio Shinoda, Kazuhiko Ishida, Eun-Gyoung Oh, Kazuhiro Sasahara, Shin ...
    1993 Volume 37 Issue 5 Pages 405-409
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    Some properties and mechanism of action of a hemolysin (VMH) produced by an enteropathogenic Vibrio mimicus strain was examined. VMH was heat-labile and inhibited by addition of divalent cations, including Ca2+, Mg2+ and Mn2+. The hemolysis by VMH was inhibited by incubating with gangliosides, suggesting that the ganglioside was the binding site on the erythrocyte membrane for VMH. Existence of a galactose moiety on reducing end of the ganglioside molecule and a sialic acid on the galactose moiety was suggested to be important for the binding of VMH molecule. Colloid osmotic manner of the hemolysis by VMH was demonstrated.
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  • Keinosuke Okamoto, Yoshio Fujii, Noriko Akashi, Shunji Hitotsubashi, H ...
    1993 Volume 37 Issue 5 Pages 411-414
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    Stock strains of Eschericia coli isolated from patients with traveller's diarrhea were examined for production of heat-stable enterotoxin II (STII). Of 400 strains examined, 3 were found to produce STII. The nucleotide sequence of the STII gene of these human strains was shown to be identical to that of porcine strains. Cultured cells of these strains induced fluid accumulation in ligated mouse intestinal loops and the activity was neutralized by anti-STII antiserum. These results suggest that STII-produciing enterotoxigenic E. coli can cause human diarrhea.
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  • Satoshi Fujimoto, Mahito Masuda, Etsuro Ono, Hiroshi Kida, Yukio Shimi ...
    1993 Volume 37 Issue 5 Pages 415-418
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    A feline T-lymphoblastoid cell line susceptible to feline immunodeficiency viruses (FIV) was established and designated as Yu-1 cells. Yu-1 cells continued to grow over one year with more than 100 successive passages in the presence of human recombinant interleukin-2. Surface antigens of Yu-1 cells were feline Pan-T+, CD4+, and CD8-. Susceptibility of Yu-1 cells to FIV strains were higher than that of the primary culture of the feline peripheral blood mononuclear cells, indicating that this cell line should be useful for isolation, propagation, and neutralization test of FIV.
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  • Eiman M. Zytoon, Hussein I. El-Belbasi, Takeo Matsumura
    1993 Volume 37 Issue 5 Pages 419-421
    Published: 1993
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    Female Aedes albopictus mosquitoes of the Miki strain were experimentally fed on defibrinated sheep blood containing 5×107PFU of chikungunya virus and 20, 000 microfilariae of Dirofilaria immitis per milliliter. Fully engorged mosquitoes transmitted the virus to a small percentage of the F1 progeny, but females of the F1 generation did not transmit the virus to the F2 progeny. The control mosquitoes that ingested the virus without microfilariae did not transmit the virus to their eggs, larvae, or pupae in the F1 or F2 generations. These results showed that A. albopictus of this strain that concurrently ingested the virus and microfilariae transmitted the virus by the transovarial route under experimental conditions.
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