bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of Mouse‐Human Hybridoma as a Useful Fusion Partner for the Establishment of Mouse‐Human‐Human Hybridoma Secreting Anti‐Tetanus Toxoid Human Monoclonal Antibody of IgM or IgG Class
Authors: Hirata Yuko , Sugawara Isamu 
Published: November 2013

AbstractWe succeeded in establishing a mouse‐human (M‐H) heterohybridoma clone which provides parental cells useful for human monoclonal antibody (hMoAb) production. Electron micrographs show that the M‐H hybridoma cells retain characteristics of murine origin with regard to chromatin patterns, small granules, granular endoplasmic reticulum and Golgi apparatus. The human DNA incorporated into the M‐H hybridoma is estimated to be about 1% of the total human chromosomal DNA. Mouse‐human‐human (M‐H‐H) hybridomas obtained by hybridization of the M‐H hybridoma cells with Epstein‐Barr virus(EBV)‐transfected human B cells secrete immunoglobulin (Ig) in amounts comparable to those of murine hybridomas. Also the M‐H‐H hybridomas grow in nude mice and are capable of producing ascites containing large quantities of Ig. The Ig class switching takes place in the M‐H‐H hybridomas at a much higher frequency than in the original EBV transformant and the M‐H hybridoma. Cells secreting specific monoclonal antibody of different Ig classes could be separated and concentrated by the use of fluorescence activated cell sorter (FACS).
Heterogeneity in Immunologic Functions of Rat Alveolar Macrophages—Their Accessory Cell Function and IL‐1 Production
Authors: Oghiso Yoichi 
Published: November 2013

AbstractAlveolar macrophages (AM) from normal rats were separated into 4 different density fractions by centrifugation on a discontinuous Percoll gradient. These fractionated (I–IV) AM, as well as unfractionated (UF) AM, were then tested for their capacities to regulate mitogen‐induced T cell proliferation. Concanavalin A (Con A)‐induced response of nylon wool‐passed non‐adherent splenic T lymphocytes was suppressed by addition of UF or higher density (III and IV) AM, while an intermediate density (II) AM fraction could enhance T cell response in a dose‐dependent manner. Similar effects of UF or fractionated (I–IV) AM on T cell responses were noted when the cultures were exposed in vitro to inert, non‐fibrogenic titanium dioxide (TiO2) particles. On the contrary, T cell response was sustained by addition of UF or higher density (III and IV) AM, and was also more prominently enhanced by an intermediate density (II) AM after the in vitro exposures to fibrogenic dust particles, like silica and asbestos. Higher interleukin 1 (IL‐1) activity was detected from these silica‐ or asbestos‐exposed cultures of UF and fractionated (II, III, and IV) AM. The IL‐1 activity was also highly detectable from the cultures of an intermediate density (II) AM fraction when cultures were unexposed or exposed in vitro to TiO2 particles. The Ia antigen expression on the surface of UF or fractionated (II, III, and IV) AM was elevated in the Con A‐pulsed co‐cultures, but not significantly different whether or not they were exposed in vitro to dust particles. These results may indicate the presence of heterogeneity in accessory cell functions and IL‐1 production among rat AM.
The Bactericidal Mechanisms of Polymorphonuclear Leukocytes against Bacteroides fragilis: Significance of the Oxygen‐Dependent System
Authors: Nishimura Toshiro , Sasada Masataka , Kubo Akemi , Kakita Tokio , Moriguchi Toshihiro , Uchino Haruto 
Published: November 2013

AbstractThe mechanism by which polymorphonuclear leukocytes (PMNs) kill ingested Bacteroides fragilis was examined using PMNs from patients with chronic granulomatous disease (CGD) which is an inherited disease characterized by the defect of their PMNs in oxygen‐radical generation. The phagocytosis of B. fragilis by PMNs from CGD patients was comparable to that by normal PMNs. Although CGD cells killed B. fragilis to some extent, they did so less effectively than the normal PMNS. B. fragilis was killed by a xanthine oxidase system that generates oxygen radicals. When PMNs were incubated with opsonized B. fragilis, B. fragilis triggered the release of O2‐ and H2O2 from normal PMNs. Thus, normal PMNs appear to kill B. fragilis by both oxygen‐dependent and oxygen‐independent mechanisms.
Characterization of Mesosomes of Micrococcus luteus: Isolation and Properties of Mesosomal Ribosomes, and Localization of Penicillin‐Binding Proteins in Mesosomal Membranes
Authors: Nakasone Noboru , Masuda Kuniyoshi , Kawata Tomio 
Published: November 2013

AbstractMesosomes were isolated and purified from Micrococcus luteus under hypertonic conditions throughout preparation processes. The purified mesosomal preparation was composed of closed tubules and vesicles. Electron‐dense ribosome‐like particles were observed within the isolated mesosomal vesicles by electron microscopy. The ribosome‐like particles were isolated from the purified mesosomes by a procedure involving solubilization of the membranes with detergents followed by centrifugation on a linear density gradient of sucrose. The isolated particles have a sedimentation coefficient of 70S in the presence of 10 mm Mg2+, when Mg2+ concentration was lowered to 0.1 mm, the particles were dissociated into two sub‐particles of 30S and 50S. The 70S particles had the same appearance as cytoplasmic 70S ribosome particles upon observations of negatively stained preparations. These findings indicate that mesosomal tubules contain ribosomes. The isolated mesosomal ribosomes had the ability for protein synthesis when polyuridylic acid‐directed polyphenylalanine synthesis was assayed. The sensitivity of mesosomal ribosomes to inhibitors, chloramphenicol and streptomycin, for protein synthesis was significantly lower than that of both cytoplasmic and cytoplasmic membrane‐bound ribosomes. In addition, three penicillin‐binding proteins were detected in the mesosomal membranes. One of these was localized predominantly in the mesosomal membranes and the other two were distributed almost equally in both mesosomal and cytoplasmic membranes.
Immunoreactivity of Cytomegalovirus‐Induced Fc Receptors
Authors: Mackowiak Philip A. , Marling‐Cason Margaret 
Published: November 2013

AbstractThe present studies were undertaken to characterize the affinity of CMV‐induced Fc receptors for each of the subclasses of human IgG and to define the specific region of the IgG Fc fragment interacting with such receptors. To do this, we infected confluent human embryonic lung (HEL) cell monolayers with CMV (strain AD169) and then used a double radiolabel assay to measure adherence of antibody‐coated E. coli 06 to such monolayers. Preincubating monolayers with each of the 4 subclasses of human IgG (but not IgA, IgM, or human or bovine albumin) abrogated the enhancing effect of CMV infection on adherence of antibody‐coated E. coli 06 to HEL monolayers. Pepsin‐derived, purified Fc fragments of human IgG had a similar abrogative effect. Preincubating these with staphylococcal protein A did not reduce their capacity to interfere with binding of antibody‐coated E. coli to CMV‐induced Fc receptors. These observations establish a broad range of immunoreactivity for CMV‐induced Fc receptors, that encompasses all 4 subclasses of human IgG. They also provide indirect evidence that the reaction site of CMV‐induced Fc receptors is in the CH2 domain of the Fc fragment.
Evaluation of the Experimental Pathogenicity of Some Cryptococcus Species in Normal and Cyclophosphamide‐Immunodepressed Mice
Authors: De Bernardis Flavia , Palliola Ennio , Lorenzini Rodolfo , Antonucci Giovanni 
Published: November 2013

AbstractThe pathogenic potential of distinct Cryptococcus species has been evaluated in mice rendered leukopenic by one or two injections of the potent immunosuppressive drug cyclophosphamide (Cy). Pathogenicity assessment included enumeration of viable cryptococcal cells in animal organs and histopathological observations. It was found that putatively non‐pathogenic species of Cryptococcus, in particular C. cereanus and C. albidus, showed significant lethality for Cy‐treated mice. In Cy‐immunodepressed mice, challenged with the infectious cryptococcal cells two days after pharmacological treatment, a significant decrease of LD50 (equivalent to at least one order of magnitude) was observed for all Cryptococcus species. However, the pathogenicity enhancement due to Cy immunodepression was greater with C. neoformans. In all cases, brain and kidney were the most invaded tissues as also evidenced by histopathological examination, which showed the typical cystic lesion. All the observations made point to the conclusion that the pathogenic potential, for the immunomodulated host, of Cryptococci other than C. neoformans is significant being quantitatively and not qualitatively different from that of C. neoformans, as evidenced by a similar organotropism and similar type of histological lesions in the target organs (brain and kidney).
Human Macrophage‐Activating Factors for Cytotoxicity
Authors: Higuchi Masahiro , Sugimoto Masamichi , Kobayashi Yoshiro , Osawa Toshiaki 
Published: November 2013

AbstractHuman T cell hybridoma, H3‐E9–6, that produces macrophage activating factors for cytotoxicity (MAF‐C) was prepared by somatic fusion of phytohemagglutinin‐activated peripheral blood lymphocytes with emetine/actinomycin D‐treated cloned human acute lymphocytic leukemia cells (CEM 11). The activities of the following were assayed: (1) macrophage‐activating factor for cytotoxicity of monocytes (MAF‐C 1 day), (2) macrophage‐activating factor for cytotoxicity of monocyte‐derived macrophages (MAF‐C 6 day), (3) macrophage‐activating factor for cytotoxicity of murine macrophages (MAF‐Cm), (4) macrophage‐activating factor for glucose consumption (MAF‐G), (5) macrophage‐activating factor for O2‐ formation (MAF‐O). The culture supernatant of H3‐E9–6 showed MAF‐C 1 day‐MAF‐C 6 day, MAF‐Cm, and MAF‐G activities. The MAF‐Cm activity was considerably enhanced by the addition of murine recombinant interferon gamma (rIFN‐γ). The MAF‐C 1 day activity in the H3‐E‐9–6 sup was not decreased by heat treatment (56 C, 30 min), by pH 2 treatment or by the addition of monoclonal anti‐human IFN‐γ antibody or polymyxin B. These data suggest that MAF‐C in H3‐E9–6 sup is distinct from human IFN‐γ or lipopolysaccharide (LPS).
Human Macrophage‐Activating Factors for Cytotoxicity
Authors: Higuchi Masahiro , Sugimoto Masamichi , Kobayashi Yoshiro , Osawa Toshiaki 
Published: November 2013

AbstractTwo different factors (MAF‐C I and MAF‐C II) were obtained by anion exchange chromatography of the culture supernatant of a human T‐cell hybridoma, H3‐E9–6, which produces macrophage‐activating factors for cytotoxicity (MAF‐C). These 2 factors induced the cytotoxicity of monocytes synergistically as a priming signal (MAF‐C I) and a triggering signal (MAF‐C II), respectively. On gel filtration on a column of Superose 12, MAF‐C II was eluted mainly at the void volume, whereas MAF‐C I was eluted in the fractions corresponding to approximate molecular weights of 30–300 K. On the other hand, gel filtration in the presence of sodium deoxycholate revealed that MAF‐C II has an approximate molecular weight of 40,000, but MAF‐C I was unstable under these conditions. When the activity for mouse macrophages (MAF‐Cm activity) was tested, the MAF‐C II fraction showed high MAF‐Cm activity in the presence of murine recombinant interferon gamma (rIFN‐γ), but the MAF‐C I fraction did not show MAF‐Cm activity even in the presence of lipopolysaccharide (LPS). These results suggest that MAF‐C I (priming lymphokine) has species specificity but MAF‐C II (triggering lymphokine) does not.
Temperature‐Modulated Physiological Characteristics of Candida albicans
Authors: Hazen Kevin C. , Hazen Beth W. 
Published: November 2013

AbstractDespite numerous investigations on candidiasis, definitive conclusions concerning virulence factors are few because of oftentimes confusing and contradictory results. By utilizing various physiologic tests, which include germ tube induction, inhibition of germination by a morphogenic autoregulatory substance, enzyme production, susceptibility to exogenous chemicals, and cell surface hydrophobicity, we demonstrated that such variability is due, in part, to the environmental conditions in which cells were grown in preparation for analysis. Room‐temperature grown cells were generally less sensitive to environmental perturbation and germinated more uniformly than cells grown at 37 C. The implication of these results in relation to pathogenic studies and the epidemiology of candidiasis is suggested.
The Primary Immune Response of the Green Toad (Bufo viridis) to Challenge with Crithidia fasciculata
Authors: Al‐Yaman Fadwa , Ingram George A. 
Published: November 2013

AbstractThe primary immune response of the green toad (Bufo viridis) following immunization with Crithidia fasciculata choanomastigotes was studied. Lysins, agglutinins, and antibodies detectable by enzyme‐linked immunosorbent assay (ELISA) were first detected in the sera of immunized animals one week after injection. The antibody titers increased to significant levels (P<0.01) and maximum values were reached seven weeks post‐immunization. The stimulated immunoglobulins were antigen‐specific, partially heat‐labile, sensitive to the reducing agent dithiothreitol, possessed precipitin activity, effectively fixed complement and exhibited an electrophoretic mobility similar to the γ‐globulins of human serum. On this basis, it is probable that the antibody produced during the primary response in green toads is high molecular weight IgM. Increases in serum lysozyme levels paralleled the rise of antibody titers. Overall, the lysozyme concentration increased two‐fold compared to the appropriate controls. This is the first report of the immune response in amphibians to experimental injection with protozoan parasites and the use of the ELISA technique to detect antibodies in amphibian sera.
Hydrolysis of Phosphatidylinositol 4, 5‐Bisphosphate and Increase in Cytosolic Free Ca2+‐Concentration Induced in a Human T Cell Leukemia Line, JURKAT, by Monoclonal Antibodies against the T3 Complex
Authors: Sasaki Terukatsu , Takei Takashi , Hasegawa‐Sasaki Hiroko 
Published: November 2013

AbstractThe monoclonal antibodies against the T3 complex on human T lymphocytes, anti‐Leu‐4, OKT3, and T3, induced an accumulation of inositol phosphates in a human T cell leukemia line, JURKAT, in the presence of LiCl. The monoclonal antibodies also induced an increase in the cytosolic free Ca2+‐concentration ([Ca2+]i) in JURKAT. The accumulation of inositol phosphates and the increase in [Ca2+]i were specifically induced by the monoclonal antibodies against the T3 complex. Other monoclonal antibodies against differentiation antigens on human T lymphocytes were not active in inducing these responses in JURKAT. Stimulation of JURKAT by anti‐Leu‐4 induced a rapid and immediate decrease in phosphatidylinositol 4,5‐bisphosphate [PtdIns(4,5)P2] and an increase in the 32P‐labeling of phosphatidic acid, which occurred after a short lag period. An analysis of inositol phosphates formed in the anti‐Leu‐4‐stimulated JURKAT indicated the formation of inositol trisphosphate. These results strongly suggested that the T3 complex or T3/antigen receptor (Ti) complex functions as a receptor which transduces antigen signal, presented by either antigen‐presenting cells or target cells, into the hydrolysis of PtdIns(4,5)P2. Fetal bovine serum at a dose of 1–20 μl/ml induced a marked and transient [Ca2+]i increase in JURKAT immedately after addition. However, the level of formation of inositol phosphates was very small in cells stimulated by fetal bovine serum. Fetal bovine serum induced an immediate increase in the 32P‐labeling of phosphatidic acid in JURKAT. These and other results suggested that serum increased [Ca2+]i in JURKAT by a mechanism different from that for the anti‐Leu‐4‐induced [Ca2+]i response.
Two Groups of Mycobacterium avium Complex Strains Determined According to the Susceptibility to Rifampicin and Ansamycin
Authors: Tsukamura Michio 
Published: November 2013

AbstractMycobacterium avium complex strains previously not exposed to any antituberculosis agents could be divided into two groups according to their susceptibility to rifampicin and ansamycin; one group susceptible to 80 μg/ml rifampicin and to 1.25 μg/ml ansamycin, and another resistant to these concentrations. In each group, the ratio of the minimal inhibitory concentration of ansamycin against that of rifampicin was greatly different depending on the strain. This naturally occurring resistance to rifampicin and ansamycin was frequently correlated to naturally occurring resistance to ethambutol, kanamycin, enviomycin, kitasamycin, and minocycline, but not correlated to that to isoniazid and sulfadimethoxine. Ansamycin was more active than rifampicin against M. bovis, M. kansasii, M. marinum, M. xenopi, and M. haemophilum.
A Comparative Study of Alteration in Lymphocyte Subsets among Varicella, Hand‐Foot‐and‐Mouth Disease, Scarlet Fever, Measles, and Kawasaki Disease
Authors: Yanase Yoshio , Tango Toshiro , Okumura Ko , Tada Tomio , Kawasaki Tomisaku 
Published: November 2013

AbstractChanges in the lymphocyte subsets of 13 patients with varicella, 5 with hand‐foot‐and‐mouth disease, 4 with scarlet fever, 10 with measles and 20 with Kawasaki disease were examined by immunofluorescent flow cytometric analysis using monoclonal antibodies against lymphocyte cell surface antigens. The results were compared with those of age‐matched normal controls. A significant increase in the percentage of Leu‐2a positive (Leu‐2a+) cells was shown during the early convalescence of varicella, scarlet fever and measles. A significant decrease in the percentage of Leu‐3a+ cells during the acute phase was common to all the diseases examined, and a significant decrease of Leu‐4+ cells was observed except in measles. As a result, a significant decrease in the Leu‐3a+/Leu‐2a+ ratio was common to all the diseases examined during the acute and/or early convalescent phases. Leu‐M3+ cells increased significantly in varicella, scarlet fever, and Kawasaki disease. HLA‐DR+ cells increased significantly in varicella and Kawasaki disease. No significant changes in the proportions of Leu‐7+, Leu‐10+, and 2H7+ cells were found throughout the course of all the diseases examined.
Fluorometric Enzyme‐Linked Immunosorbent Assay for the Measurement of IgE Antibody to Mite Dermatophagoides farinae
Authors: Sakaguchi Masahiro , Inouye Sakae 
Published: November 2013

AbstractWe have developed a fluorometric enzyme‐linked immunosorbent assay for measuring IgE antibody to Dermatophagoides farinae. Polystyrene microplates were coated with proteins extracted from the mites. The IgE antibody which attached to the solid‐phase antigen was detected by anti‐IgE antibody conjugated with β‐galactosidase. Four‐methylumbelliferyl‐β‐D‐galactoside was used as the enzyme substrate and the fluorescence intensity of the reaction product was measured. The antibody levels determined by this method well correlated with those determined by the radioallergosorbent test (RAST). This method is simpler and less expensive to carry out than the RAST when dealing with a large number of serum specimens for seroepidemiological studies of asthma and nasal allergy.
Overproduction of Gene Products by the Super‐High‐Copy‐Number Plasmid pNR333
Authors: Mochizuki Akihiko , Horiuchi Sankichi , Goto Nobuichi , Nakaya Rintaro 
Published: November 2013

AbstractThe plasmid pNR333 is a kanamycin‐resistant, deletion derivative of pNR113 with an extremely high copy number in Escherichia coli and in Proteus mirabilis. In order to determine the usefulness of pNR333 as a replication gene of vector, the genes encoding chloramphenicol acetyltransferase (CAT) and β‐galactosidase (β‐gal) were cloned individually into both pNR333 and other low‐copy‐number plasmids. The expression of the cloned genes was compared by measuring the specific activity of each enzyme and the amounts of the proteins produced. A hybrid plasmid pNR333‐cat expressed 53 times as much activity of CAT as the low‐copy plasmid S‐a which had a copy number of four. The lacZ gene cloned in pNR333 produced 17 times as much β‐gal as in the low‐copy‐number plasmid pNR1150. These results suggest that pNR333 is a useful vector plasmid for producing a large amount of polypeptides in E. coli hosts.
Quantitative Evaluation of Colonizing Ability of Vibrio cholerae O1
Authors: Nakasone Noboru , Iwanaga Masaaki 
Published: November 2013

AbstractA new method to evaluate the adhesive ability of Vibrio cholerae O1 was proposed. Broth cultured V. cholerae O1 and a piece of formalin‐fixed rabbit intestinal wall were incubated together in KRT buffer and the number of adhered organisms was counted under a scanning electron microscope. This method was much less laborious than other methods that have been used so far, and most significantly, constant results were obtained in repeated experiments. The adhesive properties of toxigenic V. cholerae O1 evaluated by this method correlated well with its observed experimental pathogenicity.
Antigen‐Specific T Cell Cluster Formation on Antigen‐Pulsed Macrophage Monolayers in Mice
Authors: Saizawa Mitsuyoshi , Mabuchi Ayako , Yokomuro Kozo , Kimura Yoshitami 
Published: November 2013

AbstractWe describe the quantitative measurement of antigen‐specific clusters formed by antigen‐pulsed macrophages and immunized T cells in mice. We have found the peripheral blood T cells show very little non‐specific adhesion to macrophages in mice. By using this population of lymphocytes in the peripheral blood as the source of immunized T cells, we could quantitate antigen‐specific cluster formation. On OVA‐pulsed monolayers of peritoneal exudate macrophages from normal BALB/c mice, syngeneic peripheral blood T cells from donors immunized with the same antigen develop 20–40 clusters per 1,000 macrophages, whereas the same T cells on non‐pulsed monolayers develop only 0–5 cluster‐like accumulations of cells. On antigen‐pulsed monolayers of macrophages from allogeneic (C57BL/6 or A/J) mice, clusters are developed only in the negative range (0–5/1,000 macrophages). Considering the observation by Braendstrup et al, these data seem to suggest that histocompatibility between macrophages and T cells is required to develop antigen‐specific T cell clusters on antigen‐pulsed macrophage monolayers, and that the genetic restriction of immune responsiveness may be directly expressed in this initial form of cellular interaction between antigen‐bearing macrophages and specific T cells.
Characterization of Rat T Cell Subset Antigen by Monoclonal Antibody
Authors: Yamaki Toshiaki , Uede Toshimitsu , Sugawara Yukihiro , Wada Takuro , Yamaguchi Akira , Kokai Yasuo , Kikuchi Kokichi 
Published: November 2013

Abstract6B2‐B8 T cell hybridoma cells were used to immunize mice, and immune spleen cells were fused with NS/1 myeloma cells. One clone, designated RTH‐7, reacted with 89.5% of rat thymocytes, 30.2% of rat spleen cells, and 42.3% of rat lymph node cells. The RTH‐7 reacted with a subset of rat T cells but not with B cells. Double staining analysis demonstrated that RTH‐7 stained a rat T cell subset distinct from R1‐10B5‐positive cells that were known to be equivalent to mouse Lyt‐2. It was revealed that RTH‐7 and W3/25 recognize different antigenic epitopes on the same molecule. The RTH‐7 as well as W3/25 substantially inhibited the production of interleukin 2 by cells in mixed lymphocyte reaction and the lymphocyte proliferation induced by mixed lymphocyte reaction. The RTH‐7 inhibited the lymphocyte proliferation induced by Con A whereas W3/25 failed to do so. The RTH‐7 defined antigen has a molecular weight of 53,000 under reducing condition and 47,000 under nonreducing condition. The RTH‐7 defined antigen showed a wide range of heterogeneity in pI (6.2–8.8). The associated molecule of approximate molecular weight of 27,000 was occasionally detected with the RTH‐7 defined antigen in 6B2‐B8 T cell hybridoma cells as well as peripheral T cells but not in thymocytes. Thus, RTH‐7 detects a cell surface antigen of a functional T cell subset of rat origin.