bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Monoclonal Antibodies to Pseudomonas aeruginosa Produced by EBV‐Transformed Cells
Authors: Suzuki Hidenori , Okubo Yuichi , Moriyama Mayumi , Sasaki Manami , Matsumoto Yoshiko , Hozumi Toyoharu
Published: November 2013

Numerous lymphoblastoid cell lines (LCLs) which secreted antibodies against Pseudomonas aeruginosa (all Fisher's immunotypes and Homma's immunotype 1) were established by Epstein‐Barr virus (EBV)‐transformation of lymphocytes. Five LCLs were established as long‐term culture lines and their properties were determined. These LCLs produced monoclonal antibodies to Fisher's immunotype 1 and 4 and Homma's immunotype 1, and their immunoglobulin classes were IgM, IgG, and IgA. We found that three monoclonal antibodies (G3‐1, H7‐2, and E10‐1) among them successfully protected mice from the corresponding immunotype of P. aeruginosa infection. Their protective dose (PD50) values were 0.5, 2.6, and 3.1 μg immunoglobulin/mouse. These human monoclonal antibodies against P. aeruginosa prepared by EBV‐transformation method will be a valuable aid for the treatment for severe P. aeruginosa infections.
A Serological Survey of Simian Virus 40 in Monkeys
Authors: Ichikawa Takashi , Minamoto Nobuyuki , Kinjo Toshio , Matsubayashi Nobuko , Matsubayashi Kiyoaki , Narama Isao
Published: November 2013

A serological survey of simian virus 40 (SV 40) was conducted by an immune adherence hemagglutination (IAHA) test in breeding monkeys. Of a total of 356 monkeys tested, 168 (47.2%) were seropositive. All 168 seropositive monkeys were detected from 224 monkeys which were bred or kept in Japan for a long time. In contrast, none of the 132 monkeys which were newly imported from Southeast Asia was seropositive. If a comparison was made in the same breeding place, the positive rate of 80.4% (111/138) of Japanese monkeys was significantly (P<0.01) higher than the 59.5% (25/42) among rhesus monkeys. The positive rate and the IAHA titers were higher in older age group (>5 years) but similar in male and female. These results indicated that SV 40 was highly prevalent among breeding monkeys in Japan.
Characterization of Autoantigens Relevant to Autoimmune Ophthalmitis and Thyroiditis in Mice Immunized with the Syngeneic Tissue Extracts and Klebsiella O3 Lipopolysaccharide
Authors: Yokochi Takashi , Oka Kazuyuki , Iwata Hitoshi , Miyadai Toshiaki , Kimura Yoshinobu , Kato Nobuo
Published: November 2013

The histological localization and biochemical properties of the autoantigens relevant to experimental autoimmune ophthalmitis and thyroiditis were studied using sera from mice hyperimmunized with the corresponding tissue extract of syngeneic mice and Klebsiella O3 lipopolysaccharide (KO3 LPS) as a potent adjuvant. Specific antigens were detected in the lens of the eyeball by immunofluorescence test with sera from mice in which ophthalmitis had been induced and the antigens were lenticular proteins with molecular weights (MW) of 15,000 (15K) to 25K, and 45K. The lenticular proteins with MW of 15K to 25K correspond to the subunits of crystalline. These findings clearly demonstrated that our experimental model for autoimmune ophthalmitis was classified as the lens‐induced uveitis. The colloids of the thyroid follicles and the follicular cells were markedly stained by sera from mice in which thyroiditis had been induced. One of the autoantigens detected in the thyroid gland was biochemically consistent with a thyroglobulin subunit. It was also shown that these autoantigens detected in the present study were organ‐specific but not species‐specific. The nature of autoantigens in the eye and the thyroid gland is discussed.
Proteinase Production and Pathogenicity of Candida albicans
Authors: Shimizu Katsumi , Kondoh Yuzuru , Tanaka Kenji
Published: November 2013

In order to investigate a role of proteinase in the pathogenesis of Candida infections, invasion of C. albicans strains of different proteinase activity into the chorioallantoic membrane (CAM) of developing chicks was studied. Eight strains were used after examining the inducible proteinase activity in the culture containing bovine serum albumin as the sole source of nitrogen. Six were proteinase‐producing strains (type I) and two were proteinase‐deficient ones (type II). Type I strains were subdivided into type Ia strains in which the proteinase activity persisted for a week in the in vitro culture and type Ib ones in which the enzyme activity was lost by the 7th day after inoculation. By inoculation onto CAM, the type I strains could invade the tissue in which secreted proteinase was detected on the periphery of the invading Candida cells by immunohistochemical method. At an early stage of the infection, proteinase secretion was detected on the surface of the yeast cells before their entry into the tissue. The type II strains remained on the surface of the CAM and did not invade the tissue where the secretion of the enzyme was not detected. The mortality rate of the chick embryo was not correlated with the degree of proteinase production of these strains. Two type Ib strains invaded the CAM tissue and elicited some tissue reactions by the host, yielding a low mortality rate of the chick embryos. These results suggested that the secretion of proteinase was an important factor for the invasion of CAM but other factors were also involved for the pathogenicity of C. albicans.
Proteinase Production and Pathogenicity of Candida albicans
Authors: Kondoh Yuzuru , Shimizu Katsumi , Tanaka Kenji
Published: November 2013

We have studied the virulence for mice of Candida albicans strains with different proteinase activity in the culture. The mortality rates for mice infected with the type Ia strains, which secrete proteinase whose activity porsisted for a week in vitro, were higher than those infected with the type Ib strains, which secrete proteinase whose activity declined at 2 or 3 days in vitro and the type II proteinase‐deficient strains. This was substantiated by the number of colony‐forming units (CFU) recovered from kidneys of mice infected with C. albicans. In the kidney tissues of mice infected with the type Ia strains, extensive invasion by fungal cells and the secretion of proteinase were histologically demonstrated, while in those infected with the type Ib and II strains fungal cells were rarely found. However, the mice infected with the type Ib strain NUM 978 were an exception; the recovery of CFU from the kidney was high, but the animals survived longer. Histologically, Candida cells were not colonized but interspersed in the tissue. Type II strain NUM 584 was found to be moderately virulent when infected at a high dose. These observations indicate that the proteinase plays a role in type Ia strains but that other factors are involved in the type Ib or II strains for the establishment of pathogenicity of C. albicans.
Nucleotide Sequence of the pnd Gene in Plasmid R483 and Role of the pnd Gene Product in Plasmolysis
Authors: Ono Kenji , Akimoto Shigeru , Ohnishi Yoshinari
Published: November 2013

The pnd gene of R plasmid R483, like the srnB gene of the F plasmid, increases the degradation of stable RNA in Escherichia coli. The nucleotide sequence of the pnd locus was determined and compared with that of the srnB locus. The genes have open reading frames that are 54% homologous, and both have an upstream inverted repeat sequence. The pnd gene expression seems to decrease the osmotic barrier of the cytoplasmic membrane, since no plasmolytic vacuoles were formed in the cells carrying the gene when the cells were exposed to hypertonic sucrose solution. This result suggests that RNase I in the periplasm passes through the altered membrane to degrade stable RNA in the cytoplasm.
Resistance of Normal Serum IgA and Secretory IgA to Bacterial IgA Proteases: Evidence for the Presence of Enzyme‐Neutralizing Antibodies in Both Serum and Secretory IgA, and Also in Serum IgG
Authors: Kobayashi Kunihiko , Fujiyama Yoshihide , Hagiwara Keiji , Kondoh Hozumi
Published: November 2013

Normal serum IgA and secretory IgA (sIgA) of subclass IgA1 were isolated from pooled human serum and milk, respectively. They were tested for their susceptibility to bacterial IgA proteases from Haemophilus influenzae, Streptococcus pneumoniae, Neisseria gonorrhoeae, and Neisseria meningitidis that cleave IgA of only the IgA1 subclass. They were also tested for susceptibility to a novel IgA‐protease from Clostridium ramosum that cleaves IgA of the IgA1 as well as the IgA2 subclass of the A2m(1) allotype. Both normal serum IgA1 and sIgA1 exhibited resistance to most IgA proteases. The one exception was the IgA protease from C. ramosum which readily cleaved both the serum IgA1 and sIgA1 into Fab and Fc fragments. Secretory component (SC) had nothing to do with the resistance of these IgAs. The resistance of these IgAs to most of the IgA proteases was found to be due to their enzyme‐neutralizing antibody activity, since the Fab but not the Fc fragment of sIgA1 showed enzyme‐inhibitory activity against these IgA proteases. Similar enzyme‐neutralizing antibody activity was found in the pepsin‐digested normal serum IgG‐(Fab')2 fragment. These results indicate that the induction of the enzyme‐neutralizing antibodies against the bacterial IgA proteases took place not only in mucosal sIgA but also in serum IgA and IgG. No enzyme‐neutralizing antibody activity against the novel IgA‐protease of C. ramosum was detected in any immunoglobulin preparations used in the present study or in the serum of a patient who carries the IgA protease‐producing strain of C. ramosum in his feces.
Chondroitin Sulfate‐Depolymerizing Activity in Streptococcus intermedius and Other Streptococci
Authors: Osano Etsuo , Hibi Eiko , Ozeki Masami , Fujii Yoshiro , Moriyama Takafumi
Published: November 2013

The type strain (ATCC 27335) and 18 human oral isolates of Streptococcus intermedius and some other related streptococcal species were tested for chondroitin sulfate C‐depolymerizing activity employing a modified screening plate method of Smith and Willett. As the results, S. intermedius strains except for ATCC 31412 strain were found to possess this activity. Propionibacterium acnes ATCC 11828 used as a positive control strain demonstrated strong activity, whereas S. intermedius strains showed only slightly detectable activity. This finding might be interesting in view of the classification of this species as well as its pathogenicity.
Protection against Campylobacter jejuni Infection in Suckling Mice by Anti‐Flagellar Antibody
Authors: Ueki Yuji , Umeda Akiko , Fujimoto Shuji , Mitsuyama Masao , Amako Kazunobu
Published: November 2013

We obtained two monoclonal antibodies of IgM class and IgA class of immunoglobulin prepared from mouse spleen cells immunized with crude flagellar preparation, and a polyclonal antibody raised against purified flagellin monomer of Campylobacter jejuni in a rabbit. The specificity of the reaction of these antibodies for flagellar filament was confirmed by Western blotting and by immunoelectron microscopy. These antibodies caused agglutination of the bacteria and inhibited the motility of the bacteria. When a strain of C. jejuni was treated with IgM class monoclonal antibody before being inoculated into suckling mice, it reduced colonization of the intestinal tract by this bacteria. Inhibition of the colonization by IgA class monoclonal antibody was less effective than that of IgM class, and the polyclonal antibody consisting mostly of IgG class immunoglobulin was without effect.
Effect of Histamine and Antihistamines on Interleukin‐1 Production by Human Monocytes
Authors: Manosroi Jiradej , Manosroi Aranya , Vithayasai Vicharn
Published: November 2013

This study was carried out on the effect of histamine hydrochloride and its antagonists on the production of interleukin‐1 (IL‐1) by lipopolysaccharide (LPS)‐stimulated adherent human monocytes (AHM) from normal healthy blood donors. IL‐1 activity was evaluated by incorporation of [3H]‐thymidine in mouse thymocytes in samples of 1:3 dilution. The result indicated that histamine hydrochloride significantly suppressed IL‐1 production by AHM at 10−3 m and 10−10 m in 14 donors with maximal suppression observed at 10−3 m. A 1‐hr incubation with histamine hydrochloride (10−3 m) before addition of LPS was found to be appropriate. Cimetidine, an H2‐antagonist at 10−3 m, 10−5 m, and 10−7 m significantly inhibited the effect of histamine hydrochloride (10−3 m) and gave maximum inhibition at 10−5 m, whereas chlorpheniramine maleate, and H1‐antagonist had no significant inhibitory effect at the concentrations studied (10−4 m, 10−5 m, and 10−7 m). Histamine hydrochloride (10−3 m) added alone had no significant suppressive effect, while cimetidine (10−5 m) alone had a significant stimulatory effect on IL‐1 production by AHM.
Augmentation of Killer Activity of Murine Spleen Cells against Allogeneic and Syngeneic Tumor Cells by Inoculation of Alloantigen in vivo
Authors: Nishi Yasuaki , Hosokawa Tomohide , Aoike Akira , Kamahora Takeshi , Sudo Hiromasa , Kawai Keiichi
Published: November 2013

After C57BL/6 (B6) mice were inoculated with BALB/c spleen cells via tail vein, kinetics of cytotoxic activities in the B6 mice against sensitizing alloantigens (H‐2d) and against syngeneic antigens were investigated using, as target cells, P815 mastocytoma cells (H‐2d) and B16 melanoma cells (H‐2b). Cytotoxic activity against P815 in the B6 spleen cells reached a peak 3 days after alloantigen inoculation, decreased drastically on day 5 and rose again thereafter. The profile of anti‐B16 cytotoxic activity was similar to that of anti‐P815 activity. The cytotoxic activity against P815 was inhibited partially by cold B16, but that against B16 was not inhibited by cold P815. Surface phenotype of cytotoxic cells against P815 was Lyt2+, Thyl+, Asialo GM1+ and that of cytotoxic cells against B16 was Lyt2−, Thyl+/−, and Asialo GMl+. The results indicate that inoculation of B6 mice with allogeneic BALB/c spleen cells induce two types of cytotoxic cells; one is similar to lymphokine‐activated killer (LAK) cells and the other is activated natural killer cells.
An Ultrastructural Study of HeLa Cell Invasion with Salmonella typhi GIFU 10007
Authors: Yokoyama Hideaki , Ikedo Masanari , Kohbata Shunro , Ezaki Takayuki , Yabuuchi Eiko
Published: November 2013

Scanning electron micrograph of HeLa S3 monolayered cells, inoculated with viable bacteria of a Salmonella typhi strain GIFU 10007, revealed that the extended microvilli tangled the bacteria within 10 min after inoculation. The micrographs of HeLa cells, at 1 hr after inoculation, indicate the following: shortening of bacterium‐attached microvilli, subsiding of tangled bacteria into microvilli bush, and then attachment of bacterial soma to cell surface making the cell membrane depressed. The transmission electron micrographs, at 1 hr after inoculation, demonstrated the findings of interaction between HeLa cell and S. typhi 10007, similar to those observed on scanning electron micrographs. Hair‐like fine structures from the soma of challenge organisms were also observed. They were in contact with HeLa cell microvilli and cell membrane. The bacteria were first partially and then totally surrounded by the HeLa cell plasma membrane. One, two, or several bacteria with intact outer membrane were enclosed in intracytoplasmic membrane‐bound vacuoles. Fragmented vacuolar membrane was still visible around the intracellularly accumulated bacteria at 24 hr after inoculation. The viable cells of S. typhi 10007 are regarded as internalizing into HeLa cells by a process of endocytosis and to multiply within the membrane‐bound vacuoles.
High Levels of Anti‐Mitochondrial Antibodies in MRL Mice Influence of Anti‐DNA Antibodies on Anti‐Mitochondrial Antibodies Measured by an Enzyme‐Linked Immunosorbent Assay
Authors: Ohshio Gakuji , Furukawa Fukumi , Yoshioka Hideyuki , Taki Yoshiro , Manabe Tadao , Ozawa Kazue , Tobe Takayoshi , Hamashima Yoshihiro
Published: November 2013

We developed an easy enzyme‐linked immunosorbent assay (ELISA) for anti‐mitochondrial antibodies, and detected high levels of anti‐mitochondrial antibodies in MRL/Mp‐lpr/lpr (MRL 1/1) mice. The influence of the presence of anti‐DNA antibodies in the tested sera on this assay was also evaluated. The monoclonal anti‐DNA antibodies, which were made from non‐immunized MRL 1/1 mice, did not react with the mitochondrial antigens adhering to polystyrene plates. Absorption with DNA had little effect on the levels of mitochondrial antibodies in MRL 1/1 sera. Our assay for mitochondria‐binding antibody was very easy and sensitive, although it could not differentiate among heterogeneous anti‐mitochondrial antibodies.
Association of HLA DR1 with High d‐Penicillamine Binding to Monocytes in Females
Authors: Watanabe Hiroshi , Kelly Heath , Dawkins Roger L.
Published: November 2013

Binding of d‐Penicillamine (d‐Pen) to human monocytes was examined by flow cytometry with fluorescent d‐Pen conjugate. Cells from HLA DR1‐positive healthy females bound significantly more d‐Pen than cells from DR1‐negative healthy females (P=0.015), and DR1 was associated with the highest binding among HLA DR antigens. In contrast, DR4 was associated with the lowest binding in healthy females. A difference in d‐Pen binding between healthy females who were DR1‐positive, DR4‐negative and those who were DR1‐negative, DR4‐positive was statistically significant (P=0.026). Neither healthy females nor healthy males showed significant associations of d‐Pen binding with HLA A, B, or C antigens, nor did healthy males show an association between strength of d‐Pen binding and any DR antigens.
Isolation and Characterization of Forespores from Bacillus megaterium
Authors: Watabe Kazuhito , Koshikawa Tomihiko , Yamazoe Mari , Ogawa Setsuko , Torii Mitsuo
Published: November 2013

A procedure for isolation of intact forespores from sporulating Bacillus megaterium cells was developed. The cells were digested with lysozyme and made to release free forespores from the protoplasts by disruption of the cytoplasmic membrane with sonication in phosphate buffer containing 10% glycerol. The suitability of the procedure was confirmed by recovery of dipicolinic acid in the isolated forespores and an electron microscopic observation. The fine structure of the forespores prepared at 6 hr (t6) after initiation of sporulation was similar to that of mature spores, except that the cortex layer and primordial cell wall were thinner and the core was larger. The density, determined by density gradient centrifugation, of the forespores isolated at t6, t10, t12, and mature spores was estimated to be 1.2783, 1.2875, 1.2861, and 1.2858, respectively. The isolated forespores at t6 and t8 were extremely heat labile (D80 of 9.5 and 21.5 min, respectively) relative to mature spores (D80 of 277.8 min). These forespores were also less resistant to organic solvents. Germination of the forespores as well as mature spores was induced by KNO3, d‐glucose, and l‐leucine. Forespores at t6 were more sensitive to KNO3‐induced germination than those at t10, t12, and mature spores when measured by reduction in the optical density of cell suspension.
A Simple Metachromatic and Fluorescent Staining Method for Microorganisms Using Carbocyanine Dye
Authors: Umeda Masaru
Published: November 2013

The cationic carbocyanine dye, 1‐ethyl‐2‐[3‐(1‐ethylnaphtho[1, 2d]‐thiazolin‐2‐ylidene)‐2‐methylpropenyl]‐naphtho[1, 2d]thiazolium bromide, interacts with several classes of anionic polymers, exhibiting metachromasia. We were able to stain various kinds of microorganisms with this dye. Gram‐negative bacteria were stained reddish purple, while Gram‐positive bacteria were stained violet or bluish purple. Stains of molds were of various colors. Yeast vegetative cells were stained reddish purple, but zygotic asci were bluish. Chlamydia trachomatis inclusions, which are surrounded by cytoplasmic membranes, were also stained red. Microorganism and cell stains have different features and can be identified also by use of fluorescent microscopy. The new staining method we report here is rapid and simple enough for routine microscopical examinations of smears of clinical specimens including microorganisms.
Detection of Cross‐Reactive Antibody to BLV p24 in Sera of Human Patients Infected with HTLV
Authors: Onuma Misao , Tsukiyama Kyoko , Ohya Ken‐Ichi , Morishima Yasuo , Ohno Ryuzo
Published: November 2013

For detection of antibody to bovine leukemia virus (BLV) major core protein of p24 and cross‐reactive antibody in human patients infected with human T cell leukemia virus type I (HTLV‐I), monoclonal antibody, D432 against BLV p24 was used by competitive binding enzyme‐linked immunoadsorbed assay (ELISA). In sera from cattle with enzootic bovine leukosis (EBL) which were positive for BLV antibodies by immunodiffusion test, 109 out of 112 (97.3%) were positive for BLV p24 antibody by competitive binding ELISA. By using the same procedures, 21 samples from adult T cell leukemia (ATL) patients and healthy carriers with HTLV‐I were tested for cross‐reactive antibody to BLV p24. All 21 samples were positive for HTLV‐I antibodies by immunofluorescence test and/or ELISA. By competitive binding ELISA using non‐treated BLV antigens, none of these 21 samples inhibited the binding of the D432. When the BLV antigen was treated by several different denaturation procedures, several HTLV‐I positive samples showed the inhibition of the D432 binding and the most effective treatment was by 2‐mercaptoethanol (2‐ME). Sixteen out of 21 samples showed the presence of cross‐reactive antibody against 2‐ME‐treated BLV antigens. The cross‐reactivity of human sample to BLV p24 antigen was further confirmed by Western blotting of the 2‐ME‐treated BLV antigens. None of the 28 samples from leukemia patients other than ATL which were negative for HTLV‐I antibodies showed inhibition of the D432 by the competitive binding ELISA.
Modulation of IgE Synthesis by IgE‐Binding Factors Released by T Cells of Asthmatic Patients with Elevated Serum IgE
Authors: Yanagihara Yukiyoshi , Kajiwara Keiichi , Kiniwa Mamoru , Kamisaki Toshiaki , Yui Yasuo , Shida Takao , Delespesse Guy
Published: November 2013

The culture supernatants of unstimulated T cells (TCS) from asthmatic patients with elevated serum IgE were tested for IgE‐binding factors (IgE‐BFs) displaying the IgE‐potentiating activity. The IgE‐BFs were detected by their ability to inhibit the rosetting of RPMI 8866 cells with ox erythrocytes coupled with mouse monoclonal antibody (E‐Mab) specific to Fc receptors for IgE (FcεR). TCS showing the rosette‐inhibiting activity significantly enhanced the spontaneous IgE synthesis by B cells of allergic individuals. Interestingly, rosette‐inhibiting factors could be removed by absorption with IgE‐Sepharose from which they were subsequently eluated with acid buffer, indicating that the rosette inhibition was indeed mediated by IgE‐BFs. In addition, such IgE‐BFs had affinity for concanavalin A and lost their IgE‐potentiating activity after treatment with trypsin and neuraminidase. In contrast, T cells treated with tunicamycin released IgE‐suppressing factors capable of inhibiting the IgE‐potentiating activity of TCS derived from untreated T cells. On the other hand, the culture supernatants from subpopulations depleted of FcεR+ T cells but not of FcγR+ T cells contained neither rosette‐inhibiting factors nor IgE‐potentiating factors, suggesting that IgE‐BFs were released by in vivo pre‐activated FcεR+ T cells. With regard to circulating FcεR+ T cells determined by E‐Mab, they were significantly higher in asthmatic patients with elevated serum IgE (0.77 ±0.15%) than in normal subjects (0.17±0.07%) in spite of a very small proportion of T cells bearing FcεR.
Interleukin 1 Production and Accessory Cell Function of Rat Alveolar Macrophages Exposed to Mineral Dust Particles
Authors: Oghiso Yoichi , Kubota Yoshihisa
Published: November 2013

Immunostimulatory effects of respirable mineral dust particles on alveolar macrophages (AM) and T lymphocytes were tested in vitro. When rat AM were incubated with fibrogenic silica and asbestos particles, a significant interleukin 1 (IL‐1) activity was generated into the culture supernatants, whereas neither AM alone nor AM incubated with non‐fibrogenic titanium dioxide (TiO2) particles, however, produced a detectable amount of IL‐1. Interleukin 2 (IL‐2) activity, as tested with IL‐2‐dependent CTLL‐2 assays, was not detectable from all of these cultures. It was also revealed that concanavalin A‐induced proliferation of T lymphocytes was enhanced in autologous AM and nylon wool‐passed spleen cell co‐cultures incubated only with fibrogenic particles, but not with non‐fibrogenic particles. Higher IL‐1 activity was detected only from co‐cultures exposed to fibrogenic particles, whereas IL‐2 activity was almost similar among these co‐cultures. These results indicate the differences in immunostimulatory effects on pulmonary macrophages and T lymphocytes among a variety of mineral dust particles.
Leukotoxic Activity in Actinobacillus (Haemophilus) actinomycetemcomitans Isolated from Periodontal Disease Patients
Authors: Ohta Hiroyuki , Kokeguchi Susumu , Fukui Kazuhiro , Kato Keijiro
Published: November 2013

Leukotoxic activity in Actinobacillus (Haemophilus) actinomycetemcomitans isolated from patients with rapidly progressive periodontitis (RP), gingivitis (G), and juvenile periodontitis (JP), and several oral bacteria, was determined by observation of morphological changes in polymorphonuclear leukocytes (PMNs). Many A. actinomycetemcomitans isolates yielded both rough‐surfaced and umbonate‐shaped colonies (A‐type), and smooth‐surfaced and convex‐shaped colonies (B‐type), when stock cultures were streaked on agar medium. Both types of cells were identical in terms of Gram stain, cell morphology, sugar fermentation profile, nitrate reduction and cellular fatty acid composition. Sonic extracts were prepared from 32 A. actinomycetemcomitans strains isolated from patients and from 3 American Type Culture Collection (ATCC) strains. Sonic extracts from 8 isolates and 2 ATCC strains induced sphering of PMNs during a 45–50 min period of incubation at 37 C. Extracts from the other oral bacteria had no effects on PMN morphology. The sphered PMNs were found by their fluorochromatic‐negative reactions to be damaged cells. The leukotoxic substance was heat‐sensitive (56 C, 30 min), trypsin‐sensitive and did not induce sphering of PMNs at 4 C. There was no clear correlation between colony type and leukotoxicity. Among 8 leukotoxic strains, 5 were isolates from an RP patient.
Immune Response to Toxoplasma gondii—Alteration of Antigen Specificity of Peripheral Blood Leukocyte Proliferative Responses in Acute Toxoplasmosis
Authors: Yano Akihiko , Aosai Fumie , Yamashita Keizo , Sugane Kazuo , Hayashi Shigeo
Published: November 2013

Ultrasonicated Toxoplasma gondii (RH strain) tachyzoite extract was chromatographed on Sephadex G‐200, and one main peak (93,000 M.W.) and three small peaks (> 160,000 M.W., 110,000 M.W., and 20,000 M.W.) were eluted. Toxoplasma‐specific proliferative T cell responses of peripheral blood leukocytes (PBL) from a patient with acute toxoplasmosis caused by accidental injection of tachyzoites of the protozoa were sequentially examined by using these fractioned antigens. As early as one week after the accidental injection of the protozoa, significant proliferative responses of PBL could be detected. The reaction of proliferative T cells was observed occurring mainly with Fr. II antigen. Then T cells began to respond to Fr. I and III in addition to Fr. II 3 weeks after the injection. Thus, expansion of antigen specificity in Toxoplasma‐specific T cell responses was observed at the initial stage of acquired acute toxoplasmosis.
Immunogenetic Analysis of Tolerance Induction in Anti‐Alloantigen Delayed Type Hypersensitivity Responses by Portal Venous Pre‐Inoculation with Allogeneic Cells
Authors: Shimizu Jun , Ogata Masato , Qian Jia‐Hua , Sato Sohichiro , Kokudo Shuhei , Hamaoka Toshiyuki , Fujiwara Hiromi
Published: November 2013

The present study investigates some of the immunogenetic bases for tolerance of anti‐allo‐delayed type hypersensitivity (DTH) responses as induced by pre‐inoculating allogeneic cells via portal venous (p.v.) route. BALB/c mice were injected with totally allogeneic C57BL/6 or H‐2 incompatible BALB.B spleen cells via p.v. route. These mice not only failed to exhibit anti‐H‐2b DTH responses, but also abrogated the potential to generate H‐2b‐specific DTH responses as induced by the subsequent immunization with H‐2b spleen cells via subcutaneous (s.c.) route. The p.v. presensitization with allogeneic spleen cells differing at either class I or class II of major histocompatibility complex (MHC) resulted in the tolerance induction of DTH responses to the respective allogeneic class I or class II MHC antigens. Moreover, the p.v. administration of the class I‐positive allogeneic cell fraction depleted of class II‐positive component into recipients differing at both class I and class II was capable of inducing anti‐class I DTH tolerance. These results indicate that anti‐allo‐class I or class II DTH tolerance can be induced independently and that the existence of class II antigens on p.v.‐presensitized cells is not necessarily required for the tolerance induction of anti‐allo‐class I DTH response.
The Effect of Biological Response Modifiers on Chronic and Latent Murine Cytomegalovirus Infections
Authors: Okada Masanobu , Minamishima Yoichi
Published: November 2013

Host‐mediated antiviral effect of 2 biological response modifiers (BRM), OK‐432, and PS‐K, against murine cytomegalovirus (MCMV) was evaluated in chronically or latently infected mice. In the early stage of chronic MCMV infection, the BRM‐induced resistance was evidenced by decrease in infectious viruses replicated in the salivary glands and by augmented cytotoxic activity of the spleen cells against YAC‐1 cells and MCMV‐infected mouse embryonic fibroblasts (MEF). In the late stage of chronic MCMV infection, the BRM treatment did not eliminate MCMV from the mice, but did prevent exacerbation of MCMV infection in the salivary glands induced by administration of cyclophosphamide (CY). In mice latently infected by MCMV, BRM treatment suppressed CY‐induced reactivation of MCMV in the salivary glands. It was suggested that the antiviral effect of BRM against MCMV in chronically or latently infected mice was based on activation of natural killer (NK) cells and cytotoxic T lymphocytes (CTL).
Increase in Intradermal Vascular Permeability Caused by Pertussis Toxin from Bordetella pertussis
Authors: Sakuma Shin , Imagawa Yoshitaka , Tokunaga Eiji , Ohtomo Nobuya
Published: November 2013

Rabbits that were injected intradermally with pertussis toxin (PT), produced from Bordetella pertussis, showed slight edema and erythema at the injection sites, but not hemorrhage nor necrosis. The edema lesions were stained blue by the intravenous injection of Pontamine Sky Blue 6B dye, suggesting that PT caused increased vascular permeability, similarly to the permeability factor (PF) of cholera toxin. The reaction of the PF of PT could be determined by measuring the diameter of the blue area. The diameter of the blue area bore a good linear relationship to the logarithm of the dose of PT. The activity of the PF was neutralized by anti‐PT rabbit serum. Detoxification of PT with formalin did not increase the vascular permeability, but reverted pertussis toxoid showed a PF reaction in proportion to the reverted leukocytosis‐promoting and histamine‐sensitizing activities of PT. The supernate of a Bordetella pertussis culture also induced a PF reaction and the reaction could be made clear by heating the supernate at 56 C for 30 min, but the supernate of Bordetella bronchiseptica did not induce the reaction at all.
Human T‐Lymphoblastoid Cell Lines with High and Low Abilities to Produce Interferon‐Gamma Constitutively and Their Susceptibilities to Interferon
Authors: Matsuyama Masako , Sugamura Kazuo , Hinuma Yorio
Published: November 2013

A human T‐lymphoblastoid cell line, TCL‐Fuj, produces large amounts of interferon (IFN)‐γ constitutively. A variant cell line, 2M, was derived from it. Both cell lines express similar surface antigen markers, but differ in surface morphology. Compared with the parent TCL‐Fuj cell line, 2M produced less IFN‐γ constitutively but more in response to IFN inducers. The IFNs produced constitutively and on stimulation with inducers were analyzed by SDS‐polyacrylamide gel electrophoresis. In TCL‐Fuj cells, the constitutive and induced IFNs consisted of the same molecular species (22K and 39K). In 2M cells, smaller IFNs were produced constitutively (18K and 32K) and induction resulted in a marked increase of 22K molecules. These two cell lines also differed in sensitivity to the antiviral activity of IFN. Other T‐lymphoblastoid cell lines, HPB‐ALL and TCL‐Fuj 4 cells, which did not produce IFN‐γ were permissive for vesicular stomatitis virus (VSV) replication; its growth was markedly suppressed by IFN‐γ and ‐α. TCL‐Fuj cells were also permissive for VSV, but were not susceptible to the antiviral effect of the IFNs. In contrast, in 2M cells the multiplication of VSV was restricted; the viral yield was further reduced by the IFNs and increased by treatment with anti‐human IFN‐γ serum. Several clonal cell lines derived from TCL‐Fuj and 2M cells had characteristics similar to the respective parent cell lines. The growth of both cell lines was not affected by IFN‐γ or by ‐α. The separation of antiviral and anti‐proliferative susceptibilities was peculiar to 2M cells unlike other cell lines.
Demonstration of Cross‐Reactive Antibodies to Mycoplasmas in Human Sera by ELISA and Immunoblotting
Authors: Sasaki Tuguo , Bonissol Christiane , Stoiljkovic Bob , Ito Kenichiro
Published: November 2013

Varying levels of cross‐reactivity to some mycoplasma species were observed in the sera of patients infected with Mycoplasma pneumoniae and even in normal human sera by enzyme‐linked immunosorbent assay (ELISA). The absorption of the patients' sera with M. pneumoniae lysate showed the decrease in ELISA titers not only to M. pneumoniae, but also to other mycoplasma species. These results suggested the existence of cross‐reactive antibodies to mycoplasmas in human sera. Cross‐reactive antibodies to M. pneumoniae and other mycoplasmas in the patients' sera were also demonstrated by Western blotting technique.
Application of a New Medium Supplement for Propagation and Storage of Human Cytomegalovirus
Authors: Okada Masanobu , Minamishima Yoichi
Published: November 2013

A new medium supplement, NU‐SERUM, was evaluated for cultivation of human embryonic lung fibloblasts (HEL) and for propagation and storage of human cytomegalovirus (HCMV). NU‐SERUM was comparable to fetal bovine serum (FBS) in promoting rapid growth of HEL if they were seeded at a sufficient density. HCMV replicated quite satisfactorily in HEL cultured with media supplemented with NU‐SERUM as well as FBS. Inactivation of HCMV at 37 C occurred similarly when the medium contained FBS or NU‐SERUM. However, at —70 C, HCMV was less stable in NU‐SERUM‐containing medium than in FBS‐containing medium. Sorbitol added to the NU‐SERUM‐containing medium improved the unstableness of HCMV at —70 C, and HCMV was storable with such medium. Thus, NU‐SERUM is useful as an alternative to FBS not only for growth of HEL but also for propagation and storage of HCMV.
2‐Mercaptoethanol Acts as a Potentiating Factor of Interleukin‐2‐Dependent Lymphocyte Proliferation
Authors: Sugama Kazushige , Namba Yuziro , Hatanaka Masakazu , Hanaoka Masao
Published: November 2013

An interleukin‐2 (IL‐2) dependent cell line which could be grown continuously with crude concanavalin A‐stimulated rat or mouse spleen cell culture supernatant could not survive for over 48 hr in the culture medium supplemented with partially purified or recombinant IL‐2. The cell growth was restored by adding another factor obtained from the same crude culture supernatant. This potentiating activity which was physicochemically inseparable from serum albumin was also obtained from the culture medium containing 2‐mercaptoethanol (2‐ME) and incubated at 37 C for 24 hr without the spleen cells. By further experiments, it was demonstrated that 2‐ME itself had the potentiating activity on the IL‐2‐dependent proliferation of this cell line and cysteine mediated the activity of 2‐ME. The cells could not enter S‐phase in the absence of 2‐ME even in the presence of IL‐2.
Studies on the Classification of Bovine Enteroviruses
Authors: Urakawa Toyohiko , Shingu Masahisa
Published: November 2013

Bovine enteroviruses isolated from cattle and other ruminants in various areas of the world were classified into three distinct serotypes by cross‐neutralization tests using criteria established for the differentiation of human enteroviruses. According to Western blot analysis, however, immunodominant structural polypeptides VP1 of the viruses tested have common epitopes, recognized by antisera to each of the three serotypes. These findings indicate that non‐neutralizing epitopes on VP1 are generally conserved. It is, therefore, conjectured that bovine enteroviruses were derived from a common ancestor.
The Origin of Antibody‐Forming Cells Detected in the Bone Marrow after Thymus‐Independent Antigen‐Stimulation and Abnormality in Migration of B Cells of X‐Linked Immunodeficient CBA/N Mice
Authors: Miyake Kensuke , Hayashi Shin‐Ichi , Ono Shiro , Hamaoka Toshiyuki
Published: November 2013

The anti‐TNP IgM plaque‐forming cells (PFC) were generated in the spleen and bone marrow of non‐immunodeficient normal mice after intraperitoneal administration of TNP‐LPS. Irradiation of normal mice while shielding bone marrow completely abrogated the generation of bone marrow PFC, indicating that they are derived from extramedullary sites. The bone marrow PFC response to TNP‐LPS was low in X‐linked immunodeficient CBA/N strain mice, while the spleen response was comparable to that seen in the normal mice. To further study the basis of the deficient bone marrow PFC response in CBA/N mice, spleen cells were adoptively transferred to irradiated syngeneic mice stimulated with TNP‐LPS. While spleen cells from normal mice generated high numbers of PFC in recipient bone marrow and spleen, those from CBA/N strain mice could not generate bone marrow PFC. This result was obtained regardless of whether normal or CBA/N recipients were used. These results indicate that TNP‐LPS administration normally results in the migration of B lymphocytes from the periphery into the bone marrow and that B cells from immunodeficient CBA/N strain mice bear an inherent defect in this migratory function. This migratory defect was shown to be X‐linked, as are the other previously reported B cell defects in this inbred mouse strain. The possible relationship between this migratory defect and the maturational defects of B cell lineage as reported previously in CBA/N strain mice is discussed.
High Levels of IgA‐Containing Circulating Immune Complex and Secretory IgA in Kawasaki Disease
Authors: Ohshio Gakuji , Furukawa Fukumi , Khine Myint , Yoshioka Hideyuki , Kudo Hiroyuki , Hamashima Yoshihiro
Published: November 2013

Sera of patients with Kawasaki disease were studied for the levels of IgA‐containing (C3‐fixing) circulating immune complexes (IgA‐CIC), IgG‐containing (IgG‐) CIC, total IgA, secretory IgA, and complement component (C3) by means of enzyme‐linked immunosorbent assays or single radial immunodiffusion methods. There was significantly high level of IgA‐CIC, but not IgG‐CIC. The levels of total IgA, secretory IgA, and C3 were significantly elevated. Significantly high levels of secretory IgA were found in 22 (51%) of 43 patients. The proportion of secretory IgA to total IgA also increased. These abnormalities in the IgA system may play a role in Kawasaki disease.
The Detection of Two Surface Antigens on Human Lung Macrophages Using Monoclonal Antibodies
Authors: Chida Kingo , Sato Atsuhiko , Akiyama Jin‐Ichiro , Fukumoto Tetsuo , Yamashita Akira , Gordon Mark R. , Myrvik Quentin N.
Published: November 2013

Monoclonal antibodies (McAb), designated AMH1 (IgM, lambda) and AMH2 (IgG1, Kappa), against specific surface antigens of human lung macrophages were produced by the fusion of the NS‐1 plasmacytoma cell line with spleen cells from BALB/c mice immunized with bronchoalveolar lavaged (BAL) cells obtained from selected smoking subjects. The screening and characterization of these McAb were carried out employing cellular radioimmunoassay, flow cytofluorography, and immunohistochemical methods. These two antibodies specifically reacted with macrophages in the alveolar spaces and BAL fluids. AMH1 did not react with peripheral blood cells including freshly separated monocytes, cultured monocytes, lymphocytes, granulocytes, and platelets. In addition, AMH1 did not react with peritoneal exudate cells or pleural exudate cells. On the other hand AMH2 showed the dull‐positive reaction with some monocytes and pleural exudate cells among above‐mentioned cells. These two McAb seemed to detect cell surface antigens that are expressed by highly differentiated or mature macrophages compared to OKM1. These antibodies will allow not only better characterization of immune cells but also assessment of maturity of lung macrophages.
Preparation of Monoclonal Antibodies against a Glycolipid Asialo GM1
Authors: Shimada Shizuo , Iwata Daiji
Published: November 2013

Five IgM monoclonal antibodies (MAbs), MW‐1, MW‐2, MW‐3, MW‐4, and MW‐5, against a glycolipid asialo GM1 were prepared from hybridoma clones obtained by the fusion of mouse NS‐1 myeloma cells with spleen cells from a mouse immunized with asialo GM1 adsorbed to naked Salmonella. All the MAbs reacted only with asialo GM1 when their reactivities were examined by enzyme‐linked immunosorbent assay (ELISA) and thin‐layer chromatography (TLC)‐immunostaining using structurally related glycolipids. The MAbs showed a complement‐dependent lysis of mouse natural killer (NK) cells, but the lytic activities were weaker than that of a rabbit polyclonal anti‐asialo GM1 antibody. When they were mixed, the anti‐NK activity was increased to a level almost comparable to that of the polyclonal antibody. These results suggest that all the MAbs obtained are specific for asialo GM1 and that they may be different in fine specificity for the glycolipid. Significance of the MAbs in immunological and neurochemical studies is discussed.
Effect of the Two‐Year Milk‐Feeding on the Gastrointestinal Microflora of the Cynomolgus Monkey (Macaca fascicularis)
Authors: Benno Yoshimi , Honjo Shigeo , Mitsuoka Tomotari
Published: November 2013

The effect of the two‐year milk‐feeding on the gastrointestinal (GI) microflora of a cynomolgus monkey was determined. Bifidobacterium spp. in the animal fed with cow's milk alone were distributed in higher number in the stomach, duodenum, cecum, and rectum. The increasingly higher numbers of Lactobacillus spp. and Candida spp. were counted in all regions of the GI tract of the animal fed with the formula diet. Bacteroidaceae, Enterobacteriaceae, and Streptococcus spp. were detected from the upper to lower intestines in the monkey given only milk, whereas in the monkey given the formula, those bacterial species were localized in only the lower intestines.
Hemagglutination Activity and Localization of Fc Receptor of Group A and G Streptococci
Authors: Yamada Sakuo , Matsumoto Akira
Published: November 2013

The IgG‐Fc binding activity and binding sites on the cell surface of streptococci, strains AR1 (group A) and G148 (group G), and Staphylococcus aureus strain Cowan I were examined by hemagglutination (HA) and immunoelectron microscopic methods. No distinct difference was observed in the HA activity among these three strains. However, the strains differed in the distribution of Fc receptor. Cowan I cells (having protein A) were heavily covered with two layers of ferritin particles, whereas AR1 cells were heavily covered with a single, rough layer of ferritin particles. G148 cells (having protein G) were labeled with a relatively thin, rough ferritin layer. The trypsin susceptibility of the Fc receptors of the AR1 strain was much higher than that of the G148 strain. These results suggest that both streptococcal strains are distinctly different in the arrangement or in the conformation of the Fc receptor from the Cowan I strain. It is also suggested that the Fc receptor molecules of the streptococcal strains differ from each other not only in conformation but also in trypsin susceptibility.
Expression of Human Cytomegalovirus Immediate Early Antigens Is Responsible for a Novel Chromatin Structure of Infected Human Embryonic Lung Cells
Authors: Maeda‐Takekoshi Fumiko , Takekoshi Masataka , Ihara Seiji , Watanabe Yasushi
Published: November 2013

Whereas human embryonic lung (HEL) cells displayed chromatin fibers composed of a repeat of conventional nucleosomes of 15 nm in diameter, human cytomegalovirus (HCMV) infection induced transient appearance of a novel chromatin structure composed of a repeat of large ellipsoids of 45–65 nm x 15–30 nm with linkers of 50–60 nm long and 6–7 nm thick. Essentially the same change in chromatin structure could be induced when uninfected HEL cell nuclei were incubated in vitro with a 0.4 m NaCl nuclear extract from HCMV‐infected HEL cells expressing immediate early antigens (IEA's) or with a similar nuclear extract from NIH/3T3 cells constitutively expressing HCMV IEA's. The latter cell line was established by transformation of the mouse cells with a plasmid carrying the HCMV major immediate early and immediate early 2 genes. These results together with those of control experiments suggest that the expression of IEA's is directly or indirectly responsible for the appearance of the novel chromatin structure in HCMV‐infected HEL cells.
Characterization of a Monoclonal Antibody to Guinea Pig T Cells That Inhibits Rosette Formation of the Cells with Rabbit Erythrocytes: Similarity of the Antigen to E‐Receptor on Human T Cells
Authors: Yamashita Toshiyuki , Koyama Jiro
Published: November 2013

A monoclonal antibody, GPT‐1, was prepared by fusion of the splenic cells of mice immunized with guinea pig thymocytes with a mouse myeloma cell line. GPT‐1 completely inhibited spontaneous rosette formation of T cells with papain‐treated rabbit erythrocytes. GPT‐1 reacted with 90% of thymocytes, 70% of peripheral blood lymphocytes, and 45% of splenic lymphocytes, but not with B cells. These results indicate that GPT‐1 has pan‐T reactivity. The antibody specifically bound to a single polypeptide chain with a molecular size of 50–65 kD. The surface density of the antigen was higher on thymocytes than on peripheral T cells, suggesting that the antigen is a certain differentiation antigen on T cells. Phytohemagglutinin‐activated T cells expressed more antigen molecules than resting T cells. In addition, GPT‐1 suppressed the proliferation of T cells induced by the mitogen, indicating that GPT‐1 recognizes a T cell‐specific surface antigen which is associated with T cell activation. Based on these results, it was concluded that GPT‐1 reacts with a guinea pig T cell surface antigen which is similar to the E‐receptor protein on human T cells (CD2 molecule).
Monoclonal Antibodies to O4 Antigen of Vibrio parahaemolyticus
Authors: Ogawa Setsuko , Koshikawa Tomihiko , Watabe Kazuhito , Yamazoe Mari , Torii Mitsuo , Sakato Nobuo , Azuma Takachika , Fujio Hajime
Published: November 2013

Four monoclonal antibodies were prepared against O4 antigen of Vibrio parahaemolyticus. All the antibodies were shown to be specific for O4 antigen by agglutination with heat‐killed O‐cells of the organism and precipitation with LPS preparations. The inhibition experiments of the precipitations with various sugars and oligosaccharides suggested that the combining sites of these hybridoma antibodies were directed to an antigenic determinant structure containing →3 and →6 linked d‐glucose, d‐galactose, and N‐acetyl‐d‐galactosamine.