bluuebunny/crossref_metadata_2025 huggingface dataset:
Cooperation between Humoral Factor(s) and Lyt‐2+ T Cells in Effective Clearance of Sendai Virus from Infected Mouse Lungs
Authors: Iwai Hiroshi , Yamamoto Shigeki , Otsuka Yuko , Ueda Katsumoto
Published: November 2013

The mechanism of cooperation between the L3T4+ and Lyt‐2+ T cell subsets in effective clearance of Sendai virus from infected mouse lungs was studied by adoptive cell transfer using nude mice. Simultaneous transfer of a long‐term‐cultured Sendai virus‐specific L3T4+ T cell line with L3T4+ cell‐depleted immune spleen cell (L3T4−) fraction to infected nude mice could result in viral clearance, although single injection with either of these cells was not effective. Instead of the L3T4+ T cells, culture supernatants of the L3T4+ T cell line or concanavalin A‐stimulated mouse spleen cells and mouse serum immunized with the virus were also active in the cooperative viral clearance with L3T4− fraction. The role of the Sendai virus‐sensitized L3T4− cell fraction in cooperative viral clearance with humoral factors could be replaced by neither T cell‐deprived immune spleen cell fraction nor normal spleen cells. The 1,500 units of recombinant mouse interleukin 2 (IL‐2), which was more than 12 times the IL‐2 activity present in the supernatants of the T cell line or concanavalin A‐stimulated spleen cells, failed to clear the virus in combination with the L3T4− fraction. Monoclonal antibodies to Sendai or mouse hepatitis viruses were also effective in the cooperative antiviral activity. IL‐2 activity was not detected in these monoclonal antibodies and the mouse immune serum. Single injection of any humoral factors failed to clear the virus. These results indicate that Sendai virus‐sensitized Lyt‐2+ subset of T cells acts cooperatively with humoral factor(s) other than IL‐2 or Sendai virus‐specific antibody present in supernatants of the T cell line, of concanavalin A‐stimulated spleen cells or hybridomas, and in mouse serum immunized with the virus.
Studies on B‐Cell Memory
Authors: Hosokawa Tomohide , Aoike Akira , Hosono Masamichi , Motoi Shigehiro , Kawai Keiichi
Published: November 2013

Effects of LPS on primary and secondary antibody responses to typical TI‐2 antigens were investigated in mice. Simultaneous injection of LPS with a TI‐2 antigen showed only little adjuvant effect on the following primary antibody response to the antigen. In contrast, either a single or multiple injections of LPS, prior to the immunization with a TI‐2 antigen, significantly augmented the following primary antibody response to the antigen. LPS, however, inhibited the development of B‐cell memory to a TI‐2 antigen when administered together with the antigen. Moreover, an injection of LPS in mice, which had strong IgM and IgG B‐cell memories to a TI‐2 antigen, caused disappearance or profound reduction of the memories. The results suggest that LPS produced by gram‐negative bacteria exerts inhibitory effects on the development and continuation of B‐cell memory to bacterial infections.
Antigenic Analysis between BCG and Tumor Cells by BCG‐Monoclonal Antibodies
Authors: Sasaki Jin‐Ichi , Kitagawa Masahiko , Tamagake Takuma , Narita Satoshi , Ahsan Chowdhury Rafiqul , Lu Changlong
Published: November 2013

Antigenic relation between Mycobacterium bovis strain BCG and experimental animal tumor cells was analyzed with BCG monoclonal antibodies (BCG‐MoAbs). Four BCG‐MoAbs of 602, 603, 609, and 612 were successfully established and applied for the antigenic analysis of Meth A, RL ♂ 1, colon tumor 26 of mouse, and line 10 of guinea pig tumor. MoAb 602 showed broad reactivity against all types of tumor cells. BCG antigen(s) was clearly recognized as small granules on the tumor cell surface under the fluorescence microscope, indicating that the animal tumor cells shared the common antigen(s) with BCG.
Suppressive Effect of Interferon‐β‐Activated Natural Killer Cells on Lipopolysaccharide‐Induced B Cell Differentiation of MRL/Mp‐1pr/1pr Mice
Authors: Arai Sumio , Kasho Tadahiro , Tomita Yuko , Munakata Tetsuo , Inoue Hiroyuki , Miyazaki Teinosuke
Published: November 2013

The suppressive effects of mouse recombinant interferon‐β (IFN‐β) on B cell differentiation of MRL/Mp‐1pr/1pr (MRL/1) mouse, a model of autoimmune diseases, and C3H/H2 mice, a normal situation, were investigated. Spleen mononuclear cells were cultured in the presence of lipopolysaccharide (LPS), and the suppressive effect of IFN‐β was examined on differentiation of B cells to plaque‐forming cells (PFCs) by highly sensitive reversed hemolytic plaque assay. IFN‐β (5,000–10,000 units/ml) suppressed more than 50% of PFCs of both MRL/1 and C3H/H2 mice. This suppressive activity as well as the cytotoxicity of natural killer (NK) cells enhanced by IFN‐β was abrogated by treatment of the spleen cells with anti‐asialo GM1 antibody in the presence of complement. This suppressive activity was also abrogated by intravenous administration of 20 μl/mouse of anti‐asialo GM1 12 hr before cultivation of spleen cells. These results suggest that NK cells activated by IFN might be responsible for the immunoregulation in autoimmune diseases.
Prevalence of Cytomegalovirus Antibodies in Brazilian and Japanese Populations in the North‐East of Brazil
Authors: Linhares Maria Ieda Siqueira , De Andrade Gildete Patriota , Tateno Seiki , Eizuru Yoshito , Minamishima Yoichi
Published: November 2013

The serological status to cytomegalovirus (CMV) was examined for 616 Brazilians and 399 Japanese immigrants living in the North‐East of Brazil. The sera were screened for IgG antibodies to CMV by enzyme‐linked immunosorbent assay (ELISA). The overall prevalence of CMV antibodies was higher in the Japanese population (83.7%) than in the Brazilian population (69.8%). The seropositivity was analyzed by factors of age, sex, ethnic background, and socioeconomic status. Mother‐baby contact seems to be a factor of significance in the seroepidemiology of CMV infection.
Location on the Chromosome of the lac Gene in a Lactose‐Fermenting Salmonella litchfield Strain
Authors: Hamada Kokichi , Nakayama Yasunori
Published: November 2013

We present conclusive evidence for the chromosomal location of the lac gene in a lactose‐fermenting Salmonella litchfield strain (AO Lac+). Two Hfr strains constructed from AO Lac+ had abilities to transfer the lac gene to S. typhimurium LT2 at relatively high frequencies. Detailed characterization of the transconjugants suggested that the lac in AO Lac+ was located on the host chromosome between galE (18 min) and trpB (34 min). Transduction experiments using P22 phage showed that the lac was cotransduced with gal, but not with trpB. These results clearly indicate that the lac gene is located at a position near 18 min of the linkage map of Salmonella.
Improved Methods for Detection and Serotyping of Coagulase from Staphylococcus aureus
Authors: Hwang Soo Myung , Seki Keiko , Sakurada Junji , Ogasawara Masami , Murai Miyo , Ohmayu Suzuko , Kurosaka Kosei , Masuda Shogo
Published: November 2013

Improved methods for detection and serotyping of staphylocoagulase were concomitantly devised. We devised an improved method for detection of coagulase activity on agarose film in the same manner as single radial immunodiffusion. The amounts of reagents required for detection of coagulase on agarose film were successfully diminished by adding polyethylene glycol (PEG) to the original formula described by Boothby et al. Using microplates in another improved method for coagulase serotyping, the amount of reaction fluid required was considerably less compared with the conventional tube method. PEG was found to be also effective to increase the efficacy of coagulase serotyping. In the presence or absence of anti‐coagulase antisera, culture supernatants of staphylococcal strain grown in brain heart infusion broth were incubated with the reaction fluid containing bovine fibrinogen, rabbit plasma, 6‐amino caproic acid, polyethylene glycol 6,000. Coagulase activity was visualized as a turbid mass formed in the wells. Turbid mass formation due to coagulase activity was type‐specifically inhibited in the presence of type‐specific antisera. Detailed procedures of the methods are precisely described with some preliminary results obtained by the methods.
Purification and Characterization of Streptococcal Erythrogenic Toxin Type A Produced by Streptococcus pyogenes Strain NY‐5 Cultured in the Synthetic Medium NCTC‐135. Comparison with the Dialyzed Medium (TP Medium)‐Derived Toxin
Authors: Kamezawa Yukio , Nakahara Takako
Published: November 2013

Streptococcal erythrogenic toxin type A (ET‐A) was purified from culture filtrate of Streptococcus pyogenes strain NY‐5 grown in a chemically defined synthetic medium NCTC‐135. We succeeded in simplifying the purification procedure, and obtained a highly purified preparation of ET‐A. The purification procedure was the combination of ultrafiltration with Amicon PM‐10 and YM‐10 membranes, chromatofocusing with PBE‐94 exchanger (pH 4.0–6.0), and gel filtration through Sephacryl S‐200. The purified toxin protein showed a single band with Mr 28,000 on SDS‐PAGE and had pI 5.2 on agarose IEF. HPLC chromatography pattern of the toxin revealed one symmetric peak. The result of amino acid analysis of the toxin was in accordance with that of Gerlach et al and with Weeks and Ferretti who reported the nucleotide sequence of the spe A gene. Biological activities of the purified toxin were remarkably potent. The mitogenic activity for rabbit lymphocytes and one skin test dose in rabbit were found at the lower dose of 10 pg and 1 ng of the toxin, respectively.
Interleukin 1 Regulation of Interleukin 2 and Interferon‐γ Gene Activation in a Human Leukemic HSB.2 Subclone
Authors: Kasahara Tadashi , Yagisawa Hitoshi , Mukaida Naofumi , Matsumura Haruo , Imai Mitsunobu , Shioiri‐Nakano Kohei
Published: November 2013

The role of interleukin 1 (IL1) in causing IL2 and interferon‐γ (IFN‐γ) production and their associated gene activation has been studied in a human leukemic HSB.2 subclone. One of the subclones, HSB.2‐C5B2 clone #28, which produced only low levels of IL2 and IFN‐γ when stimulated with phytohemagglutinin (PHA) or with a combination of phorbolmyristate‐acetate (PMA) and Ca2+ ionophore, ionomycin, showed marked IL2 and IFN‐γ production in the presence of IL1. The augmentation by IL1 was observed in both dot and Northern blot analysis, indicating that IL1 regulates lymphokine genes at the pretranslational level. The kinetics of IL2 and IFN‐γ production were essentially similar for both lymphokines except that there was a faster accumulation of IFN‐γ mRNA than IL2 mRNA. In contrast to the IL2 and IFN‐γ gene activation, IL2 receptor (Tac/p55 antigen) expression was induced directly by IL1 itself as with PMA in this subclone. In these studies, IL1 action was not replaced by the drugs raising intracellular cyclic AMP (cAMP). Taken collectively, these experiments support the notion that IL1 does not trigger IL2 gene activation per se, but amplifies the preactivated lymphokine genes initiated by PMA and ionomycin, whereas IL1 can activate the IL2 receptor gene without any other known signal requirements.
Purification and Characterization of the CS2 Pili of Colonization Factor Antigen II Produced by Human Enterotoxigenic Escherichia coli
Authors: Honda Takeshi , Cakir Nedim , Arita Michiko , Miwatani Toshio
Published: November 2013

A subtype (CS2) of the colonization factor antigen II (CFA/II) of human enterotoxigenic Escherichia coli (ETEC) was studied. Analysis revealed that CS2‐possessing ETEC was predominant among isolates from traveller's diarrhea at Osaka, Japan. TH61 pili produced by a clinical strain (TH61) were purified as a native form to homogeneity by zone electrophoresis and successive column chromatographies on Sepharose 4B and Phenyl‐Sepharose CL‐4B. It was demonstrated by immunogold staining technique and bacterial agglutination test that antisera against the purified pili of strain TH61 recognized pili of both strain TH61 and strain #C91f, a control strain possessing only CS2 pili. This suggests that TH61 pili purified in this study are CS2 pili. Subunit (pilin) of the purified pili has a molecular weight of about 16,000. Strains bearing CS2 could attach to human jejunal epithelial cells, and this attachment was inhibited by pretreating the enterocytes with purified pili. These indicate that CS2 pili are a factor responsible for attachment of ETEC bearing CS2 to human intestinal cells.
Transcription Mapping of Glycoprotein I (gpI) and gpIV of Varicella‐Zoster Virus and Immunological Analysis of the gpI Produced in Cells Infected with the Recombinant Vaccinia Virus
Authors: Kato Tamako , Kitamura Kenichi , Hayakawa Yasuhiko , Takahashi Michiaki , Kojima Asato , Sato Shigeo , Yamanishi Koichi
Published: November 2013

In order to determine the transcripts of gpI and gpIV of varicella‐zoster virus (VZV), RNA was isolated from human embryonic fibroblast cells infected with VZV and Northern blot analysis was carried out using cloned DNA probes of unique short region including gpI and gpIV genes. The analysis of RNA revealed two discrete transcripts of 3.6 and 2.15 kilobases (kb) and three transcripts of 3.6, 2.9, and 1.6 kb which hybridized to DNA probes covering the gpI and gpIV region, respectively. Next, mRNAs were hybrid‐selected, translated in vitro and the polypeptide products were immunoprecipitated with antibodies against these glycoproteins. The polypeptides with a molecular weight of 70,000 (70K) and 37K which were in vitro translational products of mRNA hybrid‐selected with the DNA clone covering gpI and gpIV were detected using antibodies against gpI and gpIV, respectively. The result showed that the 70K polypeptide is presumably the translational product of 2.15 kb mRNA and the 37K polypeptide is that of 1.6 kb mRNA. DNA fragment encoding gpI or gpIV was inserted into vaccinia virus DNA and the recombinant viruses, mO74 (gpI) and mO39 (gpIV), were used for immunological analysis. In consequence, the gpI derived from cells infected with mO74 showed antigenic characteristics similar to those of gpI from VZV‐infected cells as determined from the immunoprecipitation pattern, although the molecular weight of each polypeptide was different, and antibody produced in rabbits infected with recombinant virus had a high neutralizing activity, when the reaction was performed with complement. This suggested that gpI plays an important role for protection and recovery from VZV infection.
Human Parvovirus (HPV/B19) Infection with Purpura
Authors: Shiraishi Hiroyuki , Umetsu Koji , Yamamoto Hitoshi , Hatakeyama Yoshio , Yaegashi Nobuo , Sugamura Kazuo
Published: November 2013

A 33‐year‐old man complained of purpura (petechial hemorrhage) in chelidons, poples, axillae, and bilateral chest in addition to other symptoms such as lumbago, arthralgia, muscular pain, and fever. On the next day of the onset, human parvovirus (HPV/B19) antigen and HPV/B19 DNA were detected in his serum, and twelve days later IgM antibody to HPV/B19 became detectable. This case supports the relationship between purpura and HPV/B19 infection.
Bindings of Toxic Shock Syndrome Toxin‐1 and Staphylococcal Enterotoxins A, B, and C to Rabbit Spleen Cells
Authors: Fujikawa Hiroshi , Takayama Hiroo , Uchiyama Takehiko , Igarashi Hideo
Published: November 2013

Toxic shock syndrome toxin‐1 (TSST‐1) and staphylococcal enterotoxins (SE) A, B, and C were studied on binding to rabbit spleen cells. The toxins showed remarkable mitogenic effects on the cells. Among them, SEA and TSST‐1 had much stronger mitogenic activities than SEB and SEC. Binding study showed that labeled TSST‐1 and SEA bound considerably to cells, but that labeled SEB or SEC was not observed to bind at a detectable level under the same conditions as TSST‐1 and SEA. Competitive binding analysis between toxins to cells proved that TSST‐1 and SEA clearly competed with each other in binding. Scatchard plots for TSST‐1 and SEA in binding were linear at the doses used. The Scatchard analysis for TSST‐1 and SEA gave a dissociation constant of 2.5 × 10−9 m and 7.6 × 10−8 m and the number of binding sites per cell of 5.3 × 103 and 1.0 × 105, respectively.
Characterization of Clq Found in a Patient with Hypocomplementemic Vasculitis‐Urticaria Syndrome
Authors: Sasaki Takako , Yonemasu Kunio , Matsumoto Misako , Nagaki Kazuyoshi
Published: November 2013

Clq, a subcomponent of the first complement component, of a 60‐year‐old female patient with hypocomplementemic vasculitis‐urticaria syndrome (HVUS) was characterized. The Clq‐precipitin activity (Clq‐p) could not be detected by a routine method with 0.6% agarose in 10 mm Na‐phosphate buffer containing 10 mm EDTA (pH 7.2). Hemolytic activity of her serum complement (CH50) and levels of C1 and C4 were significantly reduced at the exacerbation stage, but levels of other complement components were almost within the normal range throughout her clinical course. The specific activity of Clq at her exacerbation stage was significantly low, and its elution position on Sephacryl S‐300 column was spread toward the low molecular weight in comparison with that of normal plasma. Molecular weights of the delayed fraction of Clq were estimated to be approximately 300,000 on the Sephacryl and 440,000 by the polyacrylamide gel electrophoresis (PAGE) containing sodium dodecyl sulfate (SDS) followed by immunoblotting, respectively. On reduction of her plasma, two bands with molecular weights equivalent to those of B and C chains of the normal Clq in an approximate molar ratio of 2: 1 were immunostained. Plasma at her exacerbation stage showed only one precipitation line against anti‐human Clq‐antiserum which was completely fused with that formed between purified normal human Clq and the same antiserum. The probable structural change of the hypofunctional Clq in the case of this HVUS is discussed.
Route‐Related Variation in the Immunogenicity of Killed Salmonella enteritidis Vaccine: Role of Antigen Presenting Cells
Authors: George Anna , Shroff Khushroo E. , Rath Satyajit , Ghosh Soma Nath , Sengupta Satya Ranjan , Kamat Ramesh S.
Published: November 2013

In order to assess the role of the route of immunization on the immunogenicity of killed Salmonella vaccine, mice were immunized with killed S. enteritidis by intraperitoneal (i.p.) and intradermal (i.d.) routes. Whereas the former was non‐immunogenic, the i.d. immunization generated an excellent delayed‐type hypersensitivity response; further, i.p. immunization could even suppress the subsequent i.d. immunization. Since the peritoneal macrophages (MO) are known to be particularly low in Ia or MHC‐class II antigens, so essential for antigen presentation, the non‐immunogenicity by i.p. route was thought to be due to their poor presentation efficiency. Poly I: poly C, an interferon inducer, is known to enhance the MHC‐class II expression; hence effect of poly I: poly C treatment on the immunogenicity of the killed vaccine by i.p. route was tested and indeed the non‐immunogenicity was corrected. Poor efficiency of presentation of S. enteritidis antigen by peritoneal cells and its improvement by prior poly I: poly C treatment was further confirmed by in vitro lymphocyte transformation test using primed T cells and peritoneal cells from normal and poly I: poly C treated mice. Poly I: poly C treatment also enhanced expression of Ia antigens on peritoneal cells.
Purification and Characterization of a T Lymphocyte‐Derived Differentiation‐Inducing Factor for Human Promyelocytic Cell Line (HL‐60) and Its Relationship to Lymphotoxin
Authors: Shimizu Yukitoshi , Nakamura Takehiko , Niki Tamotsu , Hemmi Hiromichi , Sugamura Kazuo
Published: November 2013

The human T cell leukemia virus type I (HTLV‐I)‐transformed T lymphocyte cell line MT‐2 constitutively produces differentiation‐inducing factor (DIF) for the human promyelocytic leukemia cell line HL‐60. Purification and characterization of DIF derived from MT‐2 were performed here to identify T cell‐derived DIF. DIF was purified from conditioned medium of the MT‐2 cell culture with serum‐free medium by utilizing the sequential chromatographies of anion‐exchange (mono‐Q) column, gel filtration (superose‐12) column, and hydrophobic (phenyl 5PW) column and finally the sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). In the SDS‐PAGE, one major band with a molecular weight of 56,000 and one minor band with 15,000 were stained with Coomassie Brilliant Blue and both showed DIF activity after extraction from gels. DIF had an isoelectric point of 5.8. In all purification steps, DIF activity for HL‐60 and cytotoxic activity to the sublines of mouse L‐929 were not able to be separated. Further, DIF was neutralized by antibody against lymphotoxin (LT) but not by antibody against tumor necrosis factor. These results indicate that the major DIF activity derived from MT‐2 is LT.
Carotenoid Pigments of Genus Rhodococcus
Authors: Ichiyama Satoshi , Shimokata Kaoru , Tsukamura Michio
Published: November 2013

A study of carotenoid pigments of the genus Rhodococcus was carried out. According to carotenes contained, Rhodococcus species were divided into three groups: the first group of Rhodococcus luteus, R. coprophilus, R. lentifragmentus, and R. maris, which formed β‐carotene; the second group of R. equi, R. rubropertinctus, R. aichiensis, R. sputi, R. chubuensis, R. obuensis, R. bronchialis, R. roseus, R. rhodochrous, R. rhodnii, and R. terrae, which formed γ‐carotene‐like substance; and the third group of R. aurantiacus, which formed neither carotene. Other carotenoid pigments were different according to the species.
Characterization of Monoclonal Antibodies against Human Parvovirus B19
Authors: Yaegashi Nobuo , Tada Kohtaro , Shiraishi Hiroyuki , Ishii Takehisa , Nagata Kinnya , Sugamura Kazuo
Published: November 2013

Eleven hybridoma cell lines producing mouse monoclonal antibodies (mAbs) against human parvovirus B19 were established. Their specificity was as follows. Approximately 5% of fetal erythroid cells inoculated with B19 reacted with all the mAbs and with anti‐B19 positive human serum, but not with negative serum by indirect double immunofluorescence staining. All the mAbs recognized both VP‐1 (84 kDa) and VP‐2 (58 kDa) capsid proteins of B19 virions propagated in vitro and in vivo by Western blotting, and immunoprecipitated B19 virions.
A Sensitive IL‐1 Thymocyte Co‐Stimulator Assay Using a Novel β‐D‐Galactoside Specific Lectin from the Beetle, Allomyrina dichotoma
Authors: Sakai Yasuko , Kimura Seishi , Yamashita Takao , Umetsu Kazuo , Suzuki Tsuneo , Sendo Fujiro
Published: November 2013

A sensitive thymocyte co‐stimulator assay of IL‐1 using a β‐d‐galactoside specific lectin (allo A) obtained from the beetle (Allomyrina dichotoma) is reported here. Allo A stimulated [3H]thymidine uptake of mouse thymocytes in the presence of IL‐1. The allo A assay was more sensitive than the PHA or PNA− thymocyte assay, especially at low doses of IL‐1. Optimal conditions for the allo A assay were as follows: allo A, 2.5–5.0 μg/ml; whole thymocytes, 0.5–1.0 × 106 cells/well; incubation time, 72–96 hr. The assay is sensitive and convenient and can easily be performed in any laboratory.
Protective Role of Human Antibody to the Fusion Protein of Measles Virus
Authors: Sato Takeshi A. , Kohama Tomoaki , Sugiura Akira
Published: November 2013

Absorption of a pooled human gamma globulin preparation with acetone‐treated measles virus‐infected cells removed all antibodies to measles virus antigens except a portion of the antibody to the fusion (F) protein. The residual anti‐F antibody had hemolysis‐inhibiting and virus‐neutralizing activities, inhibited spread of infection through cell fusion, and was effective in protection of passively immunized mice from fatal measles encephalitis, providing evidence for the protective role of human antibody to the F protein of measles virus.
Deoxyribonucleic Acid Hybridization Studies of Exophiala dermatitidis and Exophiala jeanselmei
Authors: Masuda Mitsugi , Naka Wataru , Tajima Shingo , Harada Takashi , Nishikawa Takeji , Kaufman Leo , Standard Paul
Published: November 2013

Exophiala dermatitidis and Exophiala jeanselmei share similar morphological features and have been confused with each other. To clarify the relationship between the two fungi, we conducted a deoxyribonucleic acid (DNA)‐DNA hybridization study using a dot blot method. Between E. dermatitidis and E. jeanselmei, only a very low level of DNA relatedness was seen and it was confirmed that these two fungi are distinct species based on DNA similarity. Close correspondence of DNA from the isolates of E. dermatitidis was obtained, whereas the isolates of E. jeanselmei were divided into 6 groups according to their DNA similarity and a possibility was shown that E. jeanselmei is composed of genetically heterogeneous groups. The subdivision of the species E. jeanselmei by the DNA‐DNA hybridization method was in agreement with serotyping exoantigens. This result suggests that DNA‐DNA hybridization studies provide an excellent tool for the identification and grouping of pathogenic dematiaceous fungi.
Regulation of Thymidine Kinase Activity in Mouse L Cells Biochemically Transformed by Varicella‐Zoster Virus
Authors: Shiraki Kimiyasu , Mori Hiroyuki , Hayakawa Yasuhiko , Yamanishi Koichi , Takahashi Michiaki
Published: November 2013

Regulation of thymidine kinase (TK) activity was examined in L(O)c133 and L(H3) cells carrying varicella‐zoster virus‐TK gene. TK activity of L(O)c133 cells was similarly high in either medium but that of L(H3) cells was high in HAT medium and low in non‐HAT medium. Cell growth was well correlated with TK activities of L(O)c133 and L(H3) cells in medium conditions. Regulation of the TK gene in L cells carrying the VZV‐TK gene is discussed.
Altered Virulence of a Pleiotropic Staphylococcus aureus Mutant with a Low Producibility of Coagulase and Other Factors in Mice
Authors: Seki Keiko , Ogasawara Masami , Sakurada Junji , Murai Miyo , Masuda Shogo
Published: November 2013

The virulence of a pleiotropic Staphylococcus aureus mutant with an extremely low producibility of coagulase and other factors was investigated in mouse. A mutant strain, designated as CL‐1, showed the same LD50 and the same intrarenal proliferation as its parental strain, when the mutant organisms were inoculated in mice in high doses. The mutant organisms, however, showed a diminished intrarenal proliferation compared with its parental organisms in low doses. This mutant strain expressed a pleiotropic phenotype such as a concomitant reduction in the producibility of coagulase, α‐toxin, and Panton‐Valentine leucocidin. The total effect due to the reduction in producibility of various factors on the virulence of the mutant strain was investigated with studies on the bacterial resistance to the phagocytic activity of leucocytes. A possible role of coagulase and that of some other staphylococcal exoproteins in the pathogenesis of S. aureus were discussed.
Characteristics of Macrophage Activation by Gamma Interferon for Tumor Cytotoxicity in Peritoneal Macrophages and Macrophage Cell Line J774.1
Authors: Miyakawa Yozo , Kagaya Keiko , Watanabe Koji , Fukazawa Yoshimura
Published: November 2013

We investigated the characteristics of macrophage‐mediated tumor cytotoxicity (MTC) against Meth A target, H2O2 generation and release of effector molecule(s) for MTC, by comparing with those of peritoneal macrophages (PMP) and macrophage cell line J774.1 during stimulation with recombinant gamma interferon (IFN‐γ). In PMP, MTC was demonstrated when they were stimulated with IFN‐γ for 12 hr (short‐term stimulation) and was abrogated when they were stimulated for 48 hr (long‐term stimulation). Enhanced H2O2 generation was observed in PMP activated by long‐term stimulation followed by triggering with PMA, but not observed by triggering with Meth A cells. By contrast, whereas non‐treated J774.1 cells have already attained a definite level of MTC, a higher MTC level was demonstrated both by short‐ and long‐term stimulations. Conversely, J774.1 cells were unable to generate H2O2 at any stage of IFN‐γ stimulation followed by triggering both with PMA or Meth A cells. The time course for stimulation of PMP by IFN‐γ for release of cytotoxic factor (CF) corresponded to that for MTC by PMP, and activities of the CF released from both activated PMP and J774.1 cells also closely corresponded to those of MTC by both cells. The serological and physicochemical characteristics of CF released from both activated PMP and J774.1 cells were determined to be closely related to those of tumor necrosis factor (TNF). These results indicate that in contrast to PMP, the J774.1 cell line is free from suppression stage for MTC and CF release during stimulation with IFN‐γ. The results suggest that TNF‐like CF plays a crucial role for MTC against Meth A target, and that H2O2 is irrelevant for MTC against Meth A.
An Application of Monoclonal Antibodies to Identification of Leptospires of Serogroup Icterohaemorrhagiae Found in China
Authors: Wan‐He Zhang , Xiu‐Fen Bai , Di‐Kai Nie , Qing‐Dong Deng
Published: November 2013

For the purpose of improving the procedures of identification of leptospires, a set of 5 monoclonal antibodies with different serological reactivity against serovars of Leptospira interrogans Icterohaemorrhagiae serogroup isolated in China was developed. One hundred and eight strains isolated from epidemic fields in 5 provinces in southern China were distinctly identified into 4 serovars of Icterohaemorrhagiae serogroup by the monoclonal antibody procedure, i.e., 98 isolates were identified as serovar lai, 7 as icterohaemorrhagiae, 2 as copenhageni, and 1 as H2. Factor antiserum procedure was used at the same time as control for typing these strains and an identical result was obtained.
Mycoplasma genitalium and Mycoplasma pneumoniae Share a 67 Kilodalton Protein as a Main Cross‐Reactive Antigen
Authors: Sasaki Yuko , Shintani Miharu , Watanabe Haruo , Sasaki Tsuguo
Published: November 2013

It was demonstrated that a 67 kilodalton (kDa) protein of Mycoplasma pneumoniae is a main cross‐reactive antigen with similar molecular weight protein of Mycoplasma genitalium by Western blot analysis using monoclonal antibody to 67 kDa protein of M. pneumoniae and hyperimmune rabbit sera directed against each mycoplasma strain.
Transcriptional Control Plays an Important Role for the Production of Heat‐Labile Enterotoxin in Enterotoxigenic Escherichia coli of Human Origin
Authors: Katayama Sei‐Ichi , Ninomiya Masaki , Minami Junzaburo , Okabe Akinobu , Hayashi Hideo
Published: November 2013

The production of heat‐labile enterotoxin (LT) in 76 strains of human enterotoxigenic Escherichia coli (ETEC) varied by a factor of 100. Three ETEC strains that differ in the levels of LT production were chosen for the cloning of LT genes (toxAB) into plasmid pBR322, and the gene structure and expression were compared in E. coli HB101. The recombinant of the low LT‐producing strain produced LT at the same level as that of the moderate LT‐producing strain, but that of the high‐level producer continued to produce at a level 14–21 times higher than the others. The restriction maps of the coding regions of the cloned LT genes (toxAB) were identical, but the flanking regions were dissimilar. The content of LT mRNA per cell, examined by Northern blot analysis, was higher in the high producer than the others by 6 times. The promoter strengths of the recombinants were all alike. LT mRNA of the high producer was more stable than that of the moderate one by 1.3 times, but the difference was not large enough to explain the difference of the content of LT mRNA. It was shown that LT production can be controlled at a transcriptional step, and DNA structure of the flanking regions may be involved in the control of the LT gene expression.
Influenza C Virus Infection in Rats
Authors: Takiguchi Kazuhiko , Tashiro Masato , Nakamura Kiyoto
Published: November 2013

Four‐week‐old rats (WKA/Hkm strain) were infected intranasally with the Ann Arbor/1/50 strain of influenza C virus and examined for clinical symptoms, virus replication, and serum antibody response. Although the animals showed no definite signs of illness, the virus replicated in the nose, and the hemagglutination‐inhibiting (HI) and neutralizing antibodies were produced in their sera. When the inoculum sizes of 106.2 and 103.2 PFU were used, virus was recovered from nasal homogenates between days 1 and 10, and serum HI antibody became detectable by 10 days after infection. The rats infected with 101.2 PFU of the virus continued to shed virus until as late as day 20 without producing serum HI antibody. The amount of virus recovered from the nose was not affected significantly by either sex. age, or strain of the rat except that a slower virus growth was seen in the LE strain. It was also observed that the rats, previously inoculated with 103.2 PFU of the virus, showed no virus shedding when reinfected 7 weeks later but produced virus though in low titers when reinfected 50 to 55 weeks later. Virus was also recovered from rats once inoculated with 101.2 PFU of the virus when challenged 7 weeks later. Thus repeated infections characteristic of human influenza C can be produced in rats under the restricted conditions.
Isolation of Tumor‐Associated Antigen from Sera of Bovine Leukemia Virus‐Infected Cattle
Authors: Kitamura Ryuji , Onuma Misao , Kawakami Kazuo , Kirisawa Rikio , Kawakami Yoshimi
Published: November 2013

Tumor‐associated antigens (TAA) expressed on the surface of enzootic bovine leukemia (EBL) cells were detected and separated from sera of bovine leukemia virus (BLV)‐positive cattle using monoclonal antibody‐conjugated immunoaffinity matrix. Eluted fraction from these sera showed 3 polypeptides with molecular weights of 70K, 52K, and 30K daltons, and these polypeptides reacted with a monoclonal antibody against TAA. However, only 70K peptide was isolated from culture supernatant of EBL B‐cell line. We also tried to examine a reversed passive hemagglutination test to develop a rapid screening system of serum TAA level, but its sensitivity was below the level of detection when EBL sera was applied directly. This is the first report on the existence of tumor antigens in sera from leukemic cattle.
Chemiluminescence Response of Human Phagocytes against IgG‐Opsonized Staphylococci with and without Protein A Activities
Authors: Yamada Sakuo
Published: November 2013

The effects of immunological IgG binding to Staphylococcus aureus and IgG binding via protein A on the chemiluminescence (CL) response of human phagocytes were examined. The results obtained by enzyme immunoassay showed a clear correlation between the magnitude of the CL response and amount of IgG on protein A‐deficient HL‐87 strain. Despite no difference in protein A activity between 209P and Cowan I strains, the CL response to IgG‐opsonized 209P cells was lower than that to Cowan I cells similarly opsonized. Moreover, the CL response to opsonized HL‐87 cells was identical with that of opsonized Cowan I cells, which was a protein A‐rich parent strain of the HL‐87. The protein A activity of Cowan I cells was significantly decreased when the cells were treated with the Fc fraction of IgG before opsonization, but such a treatment did not change the phagocytic CL response. These results strongly suggest that IgG bound to protein A via its Fc portion has no effect on the phagocytic CL response and that IgG immunologically bound to S. aureus is responsible for the opsonization of the bacteria.
Relationship between Mg2+‐Induced Hexagonal Assembly of R‐Form Lipopolysaccharides and Chemical Structure of Their R‐Cores
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Ito Hideo , Naito Setsuko , Arakawa Yoshichika , Komatsu Takayuki
Published: November 2013

The relationship between formation of the Mg2+‐induced hexagonal lattice structure by R‐form lipopolysaccharides (LPS) and chemical structure of their R‐cores was investigated using different kinds of R‐form LPS from a series of mutants of Salmonella minnesota or S. typhimurium. The optimal experimental condition for formation of the hexagonal lattice structure was to suspend LPS preparations, from which cationic material was removed by electrodialysis, in 50 mM tris (hydroxymethyl) aminomethane buffer at pH 8.5 containing 10 mM MgCl2. Under this experimental condition. Rb1 LPS formed the hexagonal lattice structure with the lattice constant of 14.0 + 0.2 nm. Ra LPS, which possesses the full length of R‐core, also formed the hexagonal lattice structure but its lattice constant was larger (18.1± 0.2 nm) than that of Rb1 LPS (the lattice structure by Ra LPS was looser than that by Rb1 LPS). All the other R‐form LPS preparations tested, RcP+, RcP‐, Rd1P‐. and Re LPS, whose R‐cores are shorter than that of Rb1 LPS, did not form the hexagonal lattice structure, but formed membranous structures showing various shapes which consisted of multiple bilaycr structures. Failure to form the hexagonal lattice structure was the common feature of these kinds of R‐form LPS irrespective of temperature at which the LPS suspensions in 10 mM MgCl2‐50 mM Tris buffer were incubated. From the results of the present study it was concluded that capability of R‐form LPS to form the hexagonal lattice structure has a close correlation with the chemical structure of their R‐cores.
Detection of Varicella‐Zoster Virus DNA by Field‐Inversion Gel Electrophoresis
Authors: Arao Yujiro , Yoshida Mariko , Bat Zeng Liang , Kori Yoshifumi , Nakatsukasa Akihiro , Takei Yoji , Aoji Katsuya , Yamada Masao , Uno Fumio , Miyoshi Kaoru , Nii Shiro
Published: November 2013

A new method for detection of varicella‐zoster virus (VZV) DNA using field‐inversion gel electrophoresis (FIGE) was devised. VZV‐genomie DNA could be differentiated from the host cell DNA of human embryonic lung (HEL) fibroblasts infected with VZV under electrophoretic conditions allowing resolution of linear and double‐stranded DNAs in the 49–230 kilobase pairs (Kb) range. The detection of VZV‐genomic DNA from infected HEL cells was successful regardless of whether the VZV was a laboratory strain, live vaccine strain, or fresh isolate. Under the same electrophoretic conditions, DNA of VZV‐infected HEL cells could be clearly differentiated from DNA obtained from HEL cells infected with herpes simplex virus type 1 (HSV‐1), type 2 (HSV‐2), or human cytomegalovirus (HCMV). Furthermore, VZV genomic DNA could be detected from as small a sample as 1.9 × 104 VZV‐infected HEL cells. Finally, we could detect VZV genomic DNA from 10 samples of vesicle tissue (blister lids, each about 1‐4 mm2) and one sample of vesicle fluid (about 5 μl) obtained from patients diagnosed as having herpes‐zoster. The results of this study indicate that FIGE is a simple and promising method for the detection of VZV from clinical materials as well as infected in vitro cultured cells.
Lack of Response of Murine Peritoneal Macrophages to in vitro Activation by Muramyl Dipeptide (MDP)
Authors: Nagao Shigeki , Akagawa Kiyoko S. , Yamada Kazuo , Yagawa Katsuro , Tokunaga Tohru , Kotani Shozo
Published: November 2013

Peritoneal exudate macrophages from mice, rats, and guinea pigs were assessed using six different parameters of macrophage activation to determine whether the cells were stimulated under similar experimental conditions. Peritoneal exudate macrophages from mice, irrespective of strain, were far less responsive to a variety of in vitro stimulatory effects of N‐acetylmuramyl‐L‐alanyl‐D‐isoglutamine than those from rats or guinea pigs, while no significant differences were noted with the reactivity to stimulation by endotoxic lipopolysaccharide. We conclude that macrophage activation by MDP in vitro is species dependent.
Inverse Relationship between the Flagella Formation and Prodigiosin Synthesis in Serratia marcescens
Authors: Kobayashi Nobuharu , Ichikawa Yoichi
Published: November 2013

Treatment by polymyxin B sulfate and ethylenediaminetetraacetate separated a 40 kilodalton (kDa) protein from the nonpigmented Serratia marcescens and even from the nonpigmented bacteria of the pigmented strains, whereas the same treatment separated the 100 kDa protein associated with the pigment formation from the pigmented bacteria. Lysozyme treatment separated the 100 kDa and/or 40 kDa proteins correlated with the pigment level. The 40 kDa protein was not an outer membrane protein but a fiagellin. These results suggest that the flagella formation was inversely related with the pigment formation.
Inhibitory Effect of Ca2+ on Formation of Mg2+‐Mediated Two‐Dimensional Hexagonal Lattice Structure by an R‐Form Lipopolysaccharide from Klebsiella pneumoniae
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Arakawa Yoshichika , Ito Hideo , Naito Setsuko
Published: November 2013

When the R‐form lipopolysaccharide (LPS) from Klebsiella pneumoniae strain LEN‐111 (03‐:Kl‐), from which cationic material had been removed by electrodialysis, was suspended in 50 mM Tris buffer at pH 8.5 containing 0.1 mM or higher concentrations of MgCl2, it formed an ordered two‐dimensional hexagonal lattice structure and its center‐to‐center distance (lattice constant) depended upon the concentration of MgCl2 and reached the shortest value (14 nm) at 10 mM. In contrast, in the presence of 0.1 to 10 mM CaCl2 in place of MgCl2, the electrodialyzed LPS did not form such an ordered hexagonal lattice structure but formed an irregular network structure with a center‐to‐center distance of 19 to 20 nm. We investigated interaction of Mg2+ and Ca2+ in formation of the hexagonal lattice structure by the electrodialyzed LPS suspended in 50 mM Tris buffer at pH 8.5. When 0.1 mM or higher concentrations of CaCl2 were mixed with 1 mM MgCl2 or when 1 mM or higher concentrations of CaCl2 was mixed with 10 mM MgCl2, the electrodialyzed LPS did not form the hexagonal lattice structure of the magnesium salt type but formed the irregular network structure of the calcium salt type. In the coexistence of equimolar or higher concentrations of CaCl2 together with 1 or 10 mM MgCl2, the binding of Mg to the electrodialyzed LPS was significantly inhibited and, conversely, the binding of Ca was enhanced as compared with when MgCl2 or CaCl2 was present alone. However, the coexistence of 10 times less molar concentrations of CaCl2 did not significantly inhibit the binding of Mg to the electrodialyzed LPS. Therefore, the inhibition of formation of the Mg2+‐mediated hexagonal lattice structure of the electrodialyzed LPS by equimolar or higher concentrations of CaCl2 accompanied the inhibition of binding of Mg but that by 10 times less molar concentrations of CaCl2 did not accompany it.
Hemagglutinating Activity of Mycoplasma salivarium and Its Attachment to Sheep Red Blood Cells
Authors: Watanabe Tsuguo , Shibata Kenichiro , Yukitake Hideharu
Published: November 2013

Mycoplasma salivarium (ATCC 23064) and 10 other strains isolated from human saliva agglutinated red blood cells of rabbits and human types A and O weakly, and those of sheep (SRBC) and human type B strongly. Glycoproteins on the surface of the organism cells and N‐acetylneuraminic acid residues and some sugars on SRBC were suggested to be involved in agglutination of SRBC. Protein A‐like activity was detected in the organism cells. The organism cells were also shown to attach to SRBC in PPLO broth (Difco) supplemented with 10% horse serum, and bivalent metal ions were suggested to be involved in the attachment. The organism cells attaching to SRBC activated complement through the alternative pathway and lyzed the SRBC.
Ammonia Production as a Virulence Expression by Mycoplasma salivarium
Authors: Matsuutra Masaroh , Seto Kanichi , Watanabe Tsuguo
Published: November 2013

Rabbits were inoculated intracutaneously with M. salivarium (ATCC 23064) cells. The size of the resulting swelling was significantly larger in 1) the sites inoculated with viable cells (7.5 × 109 CFU) suspended in a medium with arginine (arginine medium) than in those inoculated with killed cells, and in 2) those inoculated with cells suspended in arginine medium than with cells suspended in arginine‐free medium. The swelling was enhanced when rabbits had previously been immunized with the organism. This effect was concluded to be due to ammonia which the organism produced by the hydrolysis of arginine through the arginine‐dihydrolase pathway.
Electron Microscope Studies on the Intracellular Growth of PL‐1 Phage of Lactobacillus casei
Authors: Watanabe Kenji , Fukuzaki Tomoharu , Shirabe Mitsuhiro , Nakashima Yukihiko , Murata Kunihiko , Kuroiwa Ataru
Published: November 2013

Ultrathin sections of the cells of Lactobacillus casei infected with or without PL‐1 phages were observed by the rapid‐freezing and substitution‐fixation method. Phage‐head‐like particles were first observed in the nuclear region. The region was seen more widely dispersed in the cytoplasm than that observed by the conventional chemical fixation method. The features of cells just broken open by the infected phages were observed by the sedimentation method devised by us. The bursting occurred in more than one place in the cells with liberation of progeny phages.
Effect of Norspermidine and Its Related Triamines on the Cell‐Free Polyphenylalanine Synthesizing System from Vibrio parahaemolyticus
Authors: Yamamoto Shigeo , Koumoto Yasuyoshi , Shikami Satoshi , Shinoda Sumio
Published: November 2013

The effect of norspermidine and its structurally related triamines on the cell‐free polyphenylalanine synthesizing system from Vibrio parahaemolyticus was examined in connection with the requirement of the system for monovalent cation. In the absence of norspermidine, the maximal incorporation of [14C]phenylalanine into hot trichloroacetic acid insoluble material was observed under ionic conditions of 12 mM Mg2+ and 50 mM NH4+. K+ could partially substitute for NH4+, but Na + could not. The addition of norspermidine to the polyphenylalanine synthetic reaction mixture not only lowered the optimal Mg2+ concentration, but it also stimulated the polyphenylalanine synthesis up to 2‐fold with no significant increase in misin‐corporation of [14C]leucine. Other triamines having one or two methylene chains more than norspermidine were also effective in eliciting these effects. Furthermore, Na+ could not support the polyphenylalanine synthesis even in the presence of norspermidine and, on the contrary, inhibited the polyphenylalanine synthesis induced by NH4+ regardless of whether norspermidine was present or not. These findings are discussed in comparison with the properties of other bacterial cell‐free systems.
Two Kinds of P‐Fimbrial Variants of Uropathogenic Escherichia coli Recognizing Forssman Glycosphingolipid
Authors: Orino Koichi , Naiki Masaharu
Published: November 2013

Various types of fimbriae on pathogenic Escherichia coli strains have been classified by their antigenicities and recognition specificities for receptors. However, the antigenicity of fimbrial proteins does not always correlate with the fimbrial recognition specificity. In this communication, the exact carbohydrate structures recognized by the fimbriae of two human uropathogenic E. coli strains, KS71 (O4) and IH11024 (O6), that have P‐fimbrial antigen, were examined. Strain KS71 showed mannose‐resistant (MR) hemagglutination (HA) of human blood group OP1 phenotype erythrocytes, and its HA was inhibited by blood group Pk antigen, Gal(α,1–4)Gal(β,1–4)Gle‐ceramide and P antigen, GalNAc(β,1–3)Gal(α,1–4)Gal(β, 1–4)Gle‐ceramide but not by Forssman antigen, GalNAc(α,1–3)GalNAc(β,1–3)Gal(α, 1–4)Gal(β,1–4)Gle‐ceramide, as previously described in many papers. The cells also showed MR HA of sheep erythrocytes, which was potently inhibited by Forssman, and weakly by P and Pk antigens. These phenomena could not be explained by the above P adhesin specificity. This adhesin was called Forssman‐like adhesin. Strain IH11024 also caused MR HA of sheep erythrocytes but not of human erythrocytes. The HA was inhibited specifically by Forssman but neither by Pk nor P antigen. This adhesin was completely different from P adhesin and Forssman‐like adhesin in recognition of the carbohydrate epitope. This adhesin, until now called a pseudo‐type of P fimbriae, was renamed Forssman adhesin.