bluuebunny/crossref_metadata_2025 huggingface dataset:
Cell‐Mediated Immunity to Mouse Adenovirus Infection
Authors: Inada Toshiki , Uetake Hisao
Published: November 2013

Migration of peritoneal exudate cells (PEC), which were prepared from mice immunized against mouse adenovirus (M‐Ad), was inhibited upon exposure to the antigenic extract of M‐Ad‐infected cells. This inhibition was shown to be blocked when infected cells or their extracts were pretreated with antiserum against M‐Ad‐induced cell surface(s) antigen(s) or with antisera against alloantigens of infected cells. Immune spleen cell‐mediated cytolysis of M‐Ad‐infected cells was also blocked in the presence of anti‐S, anti‐alloantigen or anti‐β2m serum. Immunofluorescent antibody staining of S antigen(s) was blocked when infected cells were pretreated with anti‐alloantigen or anti‐β2m serum, whereas it was not blocked when they were pretreated with anti‐mouse immunoglobulin or anti‐Thy‐1.2 serum. Conversely, immunofluoresent antibody staining of alloantigens was blocked when infected cells were pretreated with anti‐S serum. These findings indicate that S and alloantigens are associated with each other or at least located very close to each other.
Association of Mouse Adenovirus‐Induced Cell Surface Antigen(s) with Histocompatibility Antigens
Authors: Inada Toshiki , Uetake Hisao
Published: November 2013

The topological relationship on the mouse adenovirus (M‐Ad)‐infected cell surface between virus‐induced specific cell surface(s) antigens and serologically defined major histocompatibility antigens (H‐2) was analyzed by the cap formation technique. Rhodamine‐isothiocyanate (RITC)‐labeled anti‐S serum failed to stain the surface of virus‐infected lymphoid cells which were pretreated with anti‐H‐2 serum and fluorescein isothiocyanate (FITC)‐labeled anti‐mouse immunoglobulin serum (anti‐M‐Ig) to cap the appropriate H‐2 antigens. Conversely, the capping of the S antigens by pretreatment with anti‐S followed by FITC anti‐M‐Ig serum induced cocapping of H‐2 antigens. The β2 microglobulins (β2m) were also shown to be cocapped with S antigens by anti‐β2m or by anti‐S serum. The S antigens, however, did not cocap with mouse‐immunoglobulins or Thyl. 2 antigens on virus‐infected B or T lymphocytes, respectively.

To further elucidate the molecular relationship between S and H‐2 antigens, radio‐iodinated virus‐infected cells were solubilized with Nonidet P40 (NP40) and S antigens were precipitated with anti‐S serum. When the precipitates were analysed with sodium dodecyl sulfate Polyacrylamide gel electrophoresis, two major peaks were seen at positions of molecules of about 45,000 and 12,000 daltons both of which corresponded with molecules which were observed when NP40 extracts of virus‐infected or uninfected cells were precipitated with anti‐H‐2 serum. Sequential immunoprecipitation analysis of infected cell extracts showed that S antigens were coprecipitated with either H‐2K or H‐2D antigens. These results suggest that the S antigens are somehow associated with H‐2K or H‐2D antigens separately.
Purification and Some Properties of Tetanolysin
Authors: Mitsui Noriko , Mitsui Ken'ichiro , Hase Jun'ichi
Published: November 2013

Tetanolysin was purified from the culture fluid of a strain of Clostridium tetani by ammonium sulfate fractionation, acetone precipitation and repeated gel filtration. Two hemolysins with different molecular weights were separated by gel filtration, and the smaller one, tetanolysin, was further purified. The purification raised the specific activity of tetanolysin 1,050‐fold to 500 HU/μg of protein. The purified preparation gave a single, relatively broad band on polyacrylamide gel electrophoresis, in which the activity was roughly parallel with the protein concentration. However, on sodium dodecylsulfate‐gel electrophoresis it gave two bands with nearly equal amounts of proteins, showing molecular weights of 53,000 and 48,000±3,000. Furthermore, isoelectric focusing revealed four peaks of the activity whose isoelectric pHs were 6.1, 5.6, 5.3, and 6.6 in decreasing order of the activity. These findings suggest that the preparation contains four hemolysins with different pis, which are classifiable into two groups by molecular size. The preparation was completely free of tetanus neurotoxin and proteases. Tetanolysin was more strongly inhibited by cholesterol and more rapidly adsorbed onto erythrocytes than θ‐toxin of Cl. perfringens.
Effect of Oxidizing Agents and Sulfhydryl Group Reagents on Beta Toxin from Clostridium perfringens Type C
Authors: Sakurai Jun , Fujii Yoshio , Matsuura Miho
Published: November 2013

Purified beta toxin from Clostridium perfringens type C was inactivated by the oxidizing agents o‐iodosobenzoate (OIBA), oxidized glutathione, and ferricyanide, and by the sulfhydryl group reagents 5,5′‐dithio‐bis(2‐nitrobenzoic acid) (DTNB) and N‐ethylmaleimide, iodoacetamide, and iodoacetic acid, causing loss of activity in various degrees depending on the concentration used. The activity of the toxin was not influenced by exposure to 1.0 mM of p‐chloromercuribenzoate. The toxin treated by OIBA or DTNB was reactivated by incubation with 2‐mercaptoethanol and dithiothreitol. The data suggest that beta toxin contains thiol groups which are essential for the activity.
Comparison of Liquid and Agar‐Solidified Defined Media Regarding the Physiological Mechanism by which β‐2‐Thienylalanine Inhibits Growth of Escherichia, Shigella, and Salmonella Cultures
Authors: Brown Kurt J. , Tannock Gerald W. , Elliott Robert B. , Lines David R.
Published: November 2013

Growth comparisons were made, using Shigella, Escherichia, and Salmonella cultures, in liquid and agar‐solidified defined media containing β‐2‐thienylalanine (β‐2‐t). The comparisons were performed to determine the nature of growth inhibition by β‐2‐t under different physical growth conditions.

In a plate assay, with increasing β‐2‐t mixed into the agar, inhibition of Escherichia and Shigella increased. However, Salmonella cultures were not inhibited even at the highest β‐2‐t concentrations used. With β‐2‐t added to liquid cultures, however, dose‐response growth relationships were exhibited by all three genera.

The differences occurring in β‐2‐t inhibition between liquid and plate assay conditions were not due to composition of culture plates, time of challenge of cultures with β‐2‐t, availability of oxygen and associated differences in ratios of volume of media to available surface area, selection of mutants in the plate assay, or to extractable substances from the agar.

However, when β‐2‐t diffusion into the liquid medium was delayed by using agar plug diffusion cultures, a physiological mechanism was demonstrable which largely protected Salmonella cultures, but not Escherichia and Shigella cultures, from growth inhibition.
Gel to Liquid‐Crystalline Phase Transitions of Lipids and Membranes Isolated from Escherichia coli Cells
Authors: Kato Kyoko , Bito Yukio
Published: November 2013

Differential scanning calorimetry (DSC) was used to examine the relationship of the gel to liquid‐crystalline phase transition of lipids to fatty acid composition with membrane lipids and spheroplast membranes isolated from cells of a wild strain and an unsaturated fatty acid auxotroph of Escherichia coli grown under various conditions. These lipids and membranes underwent thermotropic phase transitions at different temperatures depending on the thermal properties of their constituent fatty acids. The lipid phase transition occurred at higher temperatures in biomembranes than in extracted lipids. DSC thermograms of lipids synthesized by bacterial cells which were observed at a temperature scanning rate as slow as 0.3 K min‐1 were characterized by a distinctly plain peak summit. Endothermic peaks given by samples derived from elaidic acid‐enriched cells were relatively narrow and asymmetric. The discrepancy between the transition temperatures measured with extracted lipids and with membraneous fractions, and the shape of the endothermic peaks, are discussed.
Increased Nitroblue Tetrazolium Dye Reduction by Human Neutrophils Stimulated by Interferon In Vitro
Authors: Pak Chin‐Bin , Imanishi Jiro , Kishida Tsunataro , Matsuo Akio
Published: November 2013

Human leukocyte interferon enhanced nitroblue tetrazolium dye (NBT) reduction by human neutrophils (PMNs). Increase in NBT reduction paralleled increase in interferon dose. When human leukocyte interferon was heated to 60 C or 80 C for 30 min, both the antiviral activity and the effect on NBT reduction decreased. Human leukocyte interferon neutralized with anti‐human leukocyte interferon serum showed no effect on NBT reduction. A human fibroblast interferon preparation also enhanced NBT reduction. The species dependency of interferon was shown in NBT reduction as well as in antiviral activity.
Effect of Puromycin on Guinea Pig Lymphotoxin Release and Lectin‐Induced Cellular Cytotoxicity
Authors: Sawada Jun‐Ichi , Beppu Masatoshi , Osawa Toshiaki
Published: November 2013

In order to confirm the role of guinea pig lymphotoxin (GLT) in lectin‐induced cellular cytotoxicity (LICC), the effect of puromycin, a potent enhancer of GLT activity, on the LICC to target L · P3 cells induced by phytohemagglutinin (PHA) was investigated under serum‐free conditions.

LICC was completely inhibited by puromycin, when it was added at the initiation of LICC culture, because of the inhibition of the release of GLT from the effector lymph node cells. However, LICC was markedly enhanced when puromycin was added several hours after the initiation of LICC culture. The interpretation of these facts is that GLT release can be inhibited by puromycin, but that the GLT already released exerts an enhanced cytotoxic effect on the target cells in the presence of puromycin. Enhancement of the cytotoxicity by the addition of puromycin several hours after the initiation of LICC culture was observed even after the removal of the GLT present in the supernatant, suggesting that the morphologically intact target cells were already affected by GLT in the early stages of LICC culture.
Hydrocephalus in Suckling Rats Infected Intracerebrally with Mouse Hepatitis Virus, MHV‐A59
Authors: Hirano Norio , Goto Naoaki , Ogawa Tetsuo , Ono Katsuhiko , Murakami Toshiaki , Fujiwara Kosaku
Published: November 2013

After intracerebral inoculation of mouse hepatitis virus, MHV‐A59 strain, into 3‐ to 5‐day‐old Wistar rats, some survivors at 14 days postinoculation (p.i.) were found to lack the cerebral cortex and to have an accumulation of a considerable amount of cerebrospinal fluid. The virus titer in the brain increased exponentially after inoculation, reaching a maximum 4 to 6 days p.i. when immunofluorescence revealed virus‐specific antigen within neurons in the cerebral cortex. A small amount of infectious virus was also detectable 14 days p.i. when the cerebral anomaly was evident. This brain malformation causing hydrocephalus was due to cerebral damage by viral infection.
On the Mechanism of Inactivation of Chikungunya Virus by Tannic Acid
Authors: Konishi Eiji , Hotta Susumu
Published: November 2013

The mechanism of the action of tannic acid (TA) on Chikungunya virus (CHIKV) was investigated. Both infectivity and hemagglutination (HA) activity of CHIKV were reduced by treatment with TA in vitro. Aggregation of the TA‐treated virus particles was observed by electron microscopy. The reaction was reversible, depending on the pH of the mixture. However, mere dilution of the TA‐virus mixture or addition of other protein, such as bovine serum albumin, did not restore the lost infectivity. TA also suppressed the infectivity of RNA extracted from the virus and the HA activity of the viral membrane. The affinity between TA and CHIKV structural proteins (E1, E2, and C) was determined by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis in a discontinuous buffer system and the order of affinity was found to be C>E1>E2. Specific conditions for binding of TA to each of the virus proteins were investigated.
Presence of Viral DNA Sequences in Hamster Tumors Induced by BK Virus, a Human Papovavirus
Authors: Yogo Yoshiaki , Hondo Ryo , Uchida Seijiro , Watanabe Sumie , Furuno Akemi , Yoshiike Kunito
Published: November 2013

DNA prepared from cell lines and transplanted tumors originating from five representative types of BKV‐induced hamster tumors was examined for the presence of the BKV genome by analyzing DNA/DNA reassociation kinetics. BKV DNA sequences were detected in all cases. There were only a few (1–4) copies of BKV DNA per cell in one osteosarcoma and two ventricular tumors (one choroid plexus papilloma and one ependymoma), but there were multiple (up to 150) copies in one osteosarcoma, one ventricular tumor (choroid plexus papilloma), two insulinomas, one pineocytoma, and one cerebral neuroblastoma. In some cases the number of copies of the viral DNA differed among sister cell clones derived from the same primary tumor. Apparently some tumors contained nonintegrated free viral DNA besides the integrated BKV genome.
Adjuvant Action of Capsular Polysaccharide of Klebsiella pneumoniae on Antibody Response
Authors: Yokochi Takashi , Nakashima Izumi , Kato Nobuo , Asai Junpei , Iijima Soichi
Published: November 2013

The sequence of histological changes in the regional lymph node and other lymphoid organs of mice injected with the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K) or bacterial lipopolysaccharide (LPS) was followed. Injection of CPS‐K, but not LPS, induced the following characteristic histological changes in the regional lymph node. In the early stage there was a marked decrease in the number of small lymphocytes, accompanied by the appearance of scattered fragmented nuclei and infiltration of polymorphonuclear neutrophilic leukocytes, and in the late stage there was marked proliferation of macrophage‐like cells and pyroninophilic cells. Histological changes in the thymus and spleen and changes in cell populations in the bone marrow and peripheral blood after CPS‐K injection were essentially the same as after LPS injection. Since CPS‐K has a much stronger adjuvant action on antibody response than does LPS, it is suggested that the characteristic histological changes in the regional lymph node after injection of CPS‐K are closely related to its extraordinarily strong adjuvant action.
Recognition of Heterologous Cells by Macrophages
Authors: Sano Katsuyuki , Sugimoto Masanobu , Yasuda Tomoyoshi , Egashira Yasuyuki , Yamada Masaatsu
Published: November 2013

The ability of macrophages to recognize homologous and various heterologous cells was studied in mice, rats, and guinea pigs, in terms of the in vitro phagocytosis of non‐opsonized viable thymocytes by macrophages. Mouse, rat, and guinea pig macrophages were found to phagocytize actively thymocytes from certain heterologous animals, including chickens. For instance, mouse macrophages displayed conspicuous phagocytic activities against chicken and duck thymocytes, moderate activities against guinea pig and frog thymocytes and weak activities against rat and mouse thymocytes. On the other hand, guinea pig macrophages revealed a different behaviour: they ingested only chicken thymocytes. These observations strongly suggested that mammalian macrophages possess some ability to discriminate homologous from certain heterologous thymocytes. The results, however, did not necessarily support the idea that the degree of phagocytosis is simply related to the phylogenetic distance between the animal species from which thymocytes and macrophages originated, because of the apparent exception in the mode of phagocytosis by guinea pig macrophages. Evidence demonstrating that antibodies are not involved in this phenomenon will be presented in the accompanying paper.
Aggregation of Tobacco Mosaic Virus by Sodium Chondroitin Sulfate
Authors: Sano Yoh , Inoue Hideo
Published: November 2013

The precipitation of tobacco mosaic virus by sodium chondroitin sulfate in an aqueous solution was investigated kinetically by means of turbidimetry. The virus solution became turbid after the addition of chondroitin sulfate. A threshold concentration of chondroitin, 1.33 mg/ml, was required for virus precipitation, irrespective of the virus concentration. The precipitation resulted from a mutual spatial exclusion phenomenon, leading to the separation of the virus as a crystalline phase. The dimension of chondroitin sulfate calculated at the threshold concentration agreed well with that obtained by other methods. The initial slopes and the aggregation half‐times of the virus aggregates depended on both chondroitin and virus concentrations and the former increased with the increase in concentration of each. Above the threshold concentration of chondroitin sulfate, tobacco mosaic virus aggregation was a rapid‐aggregation process and ended within 100 sec.
Isolation and Characterization of Temperature‐Sensitive Mutants of Sendai Virus
Authors: Adachi Akio , Kanda Tadahito , Shibuta Hiroshi
Published: November 2013

Sixteen temperature‐sensitive mutants of Sendai virus were isolated from mutagenized stocks (10 mutants, designated numerically) and persistently infected cultures (6 mutants, designated alphabetically).

Based on complementation tests, virion‐associated activities, thermal inactivation, and viral RNA and hemadsorbing antigen synthesis as well as virion production in chick lung embryo cells at nonpermissive temperature, these mutants were divided into seven groups as follows.

HANA group mutants (ts−5, −9, −10,−201), defective in hemagglutinin‐neuraminidase protein, complementation group I.

F group mutants (ts‐18, −108), defective in hemolytic and cell‐fusing activity, complementation group II.

Ts‐43, defective in RNA polymerase activity, complementation group III.

Ts‐23, defective in RNA polymerase activity, interfered with the other mutants in complementation tests.

Ts‐25, defective in the incorporation of hemagglutinin‐neuraminidase protein into the virion at the stage of virus assembly.

Ts‐110, belongs to F group mutants on one hand, but is considered to carry another undetermined defect.

C group (carrier culture‐borne group) mutants (ts‐a, ‐b, ‐c, ‐d, ‐e, ‐f), defective lesion not yet determined and belong to neither complementation group I nor II. Assignment of mutants in groups iv), v), vi), and vii) to complementation groups could not be achieved.
Nonspecific Stimulation of Antibacterial Resistance by a Synthetic Thymic Factor (FTS) in Mice
Authors: Matsumoto Takao , Nakano Masayasu
Published: November 2013

Treatment of mice with 0.01–1 μg of a synthetic serum thymic factor (FTS) significantly increased their resistance to lethal doses (1–5 × 106 organisms) of Salmonella typhimurium LT2 (LT2). At least seven injections of FTS were necessary for induction of resistance, since the effects of one to three injections were variable. FTS additively potentiated the protective immunity to LT2 induced by immunization with an avirulent mutant of LT2, irrespective of the structural components of the cell wall lipopolysaccharides of the mutants, which are basically correlated with their immunogenicity and virulence. Activated macrophages may have a direct role in the induction of host resistance, since peritoneal macrophages from FTS‐treated DBA/2 mice which are resistant to the doses of LT2 used could by themselves retard bacterial growth in vitro, whereas those from CBA/N mice which are relatively susceptible to LT2 did not inhibit bacterial growth without intervention of FTS‐treated syngeneic T cells.
Complement Proteins and Macrophages
Authors: Miyama Akio , Kawamoto Yasuko , Ichikawa Hidetaka , Okamoto Keinosuke , Hara Susumu , Inoue Takashi
Published: November 2013

The secretion of factor B by mouse peritoneal macrophages was found to be enhanced following in vivo or in vitro stimulation with lipopolysaccharide (LPS). The intravenous administration of LPS to mice of various strains caused an increased release of factor B but not the release of acid phosphatase by the peritoneal macrophages obtained from the stimulated mice. In vitro stimulation of cultured macrophages with LPS resulted in an enhanced secretion of both factor B and acid phosphatase. The dose‐dependent augmentation of factor B secretion by LPS was found in the macrophages from LPS‐responsive C3H/HeN mice, whereas the macrophages from LPS‐unresponsive C3H/HeJ mice did not respond to either phenol‐extracted LPS or butanol‐extracted LPS. The ability of LPS to cause the enhancement of factor B secretion by macrophages was abolished by alkali or acid treatment of LPS, indicating that its lipid A part was responsible for the observed effect.
Three Phases in Transformation of Human Lymphocytes by Epstein‐Barr Virus, as Revealed by Colony Formation in Soft Agar
Authors: Harada Shinji , Katsuki Takato , Hinuma Yorio
Published: November 2013

Human cord lymphocytes were examined for colony formation in soft agar medium at various times after infection with Epstein‐Barr virus (EBV). The transformation process could be divided into the following three sequential phases: (1) A pre‐early phase, lasting about 10 hr post infection (pi), during which the number of transformed colonies was virus dose dependent. (2) An early phase, between about 10 and 60 hr pi, during which cloning efficiency was markedly lower than that of the pre‐early phase. (3) A later phase, starting at about 60 hr pi, during which cloning efficiency was restored and the number of colony formers increased in an exponential fashion. EBV‐associated nuclear antigen‐positive cells first appeared at the beginning (12 hr pi) of the early phase and increased continuously thereafter. Increased DNA synthesis began at about 48 hr pi. These results suggest that the cloning efficiency of the infected cells changes during the transformation process, possibly reflecting altered cell sensitivity to agar.
Effect of Muramyldipeptide, a Synthetic Bacterial Adjuvant, on Enzyme Release from Cultured Mouse Macrophages
Authors: Imai Katsuyuki , Tanaka Atsushi
Published: November 2013

Monolayer cultures of macrophages obtained by peritoneal lavage of normal or thioglycollate‐stimulated mice spontaneously secreted lysosomal enzymes into the culture medium. When the elicited macrophages were cultured in the presence of muramyldipeptide (MDP), a 20–30% increase in the release of β‐glucuro‐nidase was consistently observed and the intracellular activity decreased to about 45% of that of control cells after 6–8 days' culture. A stimulatory effect of MDP on lysozyme secretion, though less profound, was also observed. In contrast, release of neither enzyme was stimulated in resident macrophages by the addition of MDP. A neutral α‐glucosidase, which has recently been found to localize also in granules of macrophages, remained inside the cells and neither its activity nor its release was affected by the addition of MDP to either type of macrophages. A large amount of lactic dehydrogenase was released only when the resident, not the elicited, macrophages were cultured for 3–4 days and then phagocytosed zymosan.
Genetic Control of Contact Hypersensitivity
Authors: Ishii Norihisa , Ikezawa Zenro , Nagai Ryukichi , Tani Kenji , Ishigatsubo Yoshiaki , Okuda Kenji
Published: November 2013

The characteristics of suppressor cells induced by 2,4‐dinitrophenyl (DNP)‐coupled syngeneic lymphocytes (syninduced suppressor cells) were studied. 2,4‐dinitro‐1‐fluorobenzene (DNFB) contact hypersensitivity was completely suppressed when the syninduced suppressor cells were transferred intravenously. These syninduced suppressor cells had surface markers of Thy‐1, FcR− and Lyt‐2+, 3+ antigens, as well as I‐J gene products on their cell surfaces. The suppression of DNFB contact hypersensitivity was abrogated when these suppressor T cells were incubated in Petri dishes coated with the DNP‐syngeneic lymphoid cell membrane, which suggests that these suppressor T cells had the specific antigen‐binding receptors on their cell surfaces.
Close Association of Virus‐Specific Cell Surface (S) and H‐2 Antigens in Ad12‐Infected and ‐Transformed Mouse Cells
Authors: Maeta Yasunobu , Hamada Chuya
Published: November 2013

A virus‐specific cell surface (S) antigen in adenovirus type 12 (Ad12)‐transformed mouse cells has been assumed to be a direct target for cytotoxic thymus‐derived lymphocytes (CTL). In this study, the spatial proximity between the S and H‐2 antigens was determined by three different methods, the proximity and co‐capping tests, and the test for blocking of CTL‐mediated lysis by anti‐H‐2 serum. In the proximity test with Ad12‐infected thymic and splenic lymphocytes, and an Ad12‐transformed line of C3H/He (H‐2k) mouse cells, anti‐H‐2k and anti‐S sera reciprocally inhibited fluorescent‐antibody staining of the opposite antigens. By contrast, anti‐Thy‐1, 2 serum as well as anti‐Ia and anti‐Ig sera failed to show any appreciable effect in this test, when paired with anti‐S serum. In addition, the S and H‐2 antigens co‐capped in the infected thymic lymphocytes, and CTL‐mediated lysis of the transformed cells was abrogated equally by treatment of cells with anti‐S and anti‐H‐2 sera. These results clearly demonstrate that there is a close proximity between the S and H‐2 antigens on the surface of Ad12‐infected and ‐transformed mouse cells.
Mitogen‐Induced DNA Synthesis in Various Mouse Strains Infected with a Large or Small Dose of Murine Leprosy Bacilli
Authors: Yamaura Noboru , Akiyama Takehisa , Nakano Takeshi
Published: November 2013

Mice of the C57BL strain have been shown to be rather resistant to infection with Mycobacterium lepraemurium, whereas C3H mice are highly susceptible. Accordingly, it seemed to be somewhat paradoxical that enhanced antibody formation coupled with a depressed state of cell‐mediated immunity as expressed by negative macrophage migration inhibition tests was observed not in C3H but in C57BL mice when they were inoculated with a large dose of murine leprosy bacilli, as reported in our previous studies.

In the present study mitogen‐induced DNA synthesis by lymph node cells was examined in 16 strains of mice which had been infected with a large or small dose of M. lepraemurium. According to the response to two kinds of T‐cell mitogens, these mouse strains could be roughly divided into three groups consisting of two polar groups represented by C57BL/6J and C3H/HeN, respectively, and one intermediate between them.

Furthermore, both humoral and cellular immune responses so far observed in C57BL and C3H mice were substantiated by DNA synthesis by lymph node cells harvested from these strains of mice and then exposed in vitro to B‐cell and T‐cell mitogens, respectively.

However, no correlation was found between mitogen‐stimulated DNA synthesis by these 16 strains of mice and their H‐2 specificity.
Analysis of Nuclear Accumulation of Influenza NP Antigen in von Magnus Virus‐Infected Cells
Authors: Maeno Koichiro , Aoki Hiizu , Hamaguchi Michinari , Iinuma Masao , Nagai Yoshiyuki , Matsumoto Toshisada , Takeura Shigeki , Shibata Motohiro
Published: November 2013

When 1–5C‐4 cells were infected with von Magnus virus derived from influenza A/RI/5+ virus by four successive undiluted passages in chick embryos, virus‐specific proteins were synthesized but production of infectious virus was inhibited. In these cells the synthesis of viral RNA was suppressed and the nucleoprotein (NP) antigen was found predominantly in the nucleus in contrast to standard virus‐infected cells in which the antigen was distributed throughout the whole cell. The intracellular location and migration of NP were determined by isotope labeling and sucrose gradient centrifugation of subcellular fractions. In standard virus‐infected cells NP polypeptide was present predominantly in the cytoplasm in the form of viral ribonucleoprotein (RNP) and intranuclear RNP was detected in reduced amounts. In contrast, in von Magnus virus‐infected cells NP polypeptide was present predominantly in the nucleus in a nonassembled, soluble form and the amount of cytoplasmic RNP was considerably reduced. After short‐pulse labeling NP was detected exclusively in the cytoplasm in a soluble form and after a chase a large proportion of such soluble NP was seen in the nucleus. It is suggested that a large proportion of the NP synthesized in von Magnus virus‐infected cells is not assembled into cytoplasmic RNP because of the lack of available RNA and the NP migrated into the nucleus and remained there.
Target Antigen of Vaccinia‐Infected Cells Recognized by Virus‐Specific Cytotoxic T Lymphocytes
Authors: Oie Masayasu , Ichihashi Yasuo
Published: November 2013

A vaccinia‐specific target antigen for recognition of anti‐vaccinia cytotoxic T lymphocytes (CTL) was found to be formed on the surface of infected cells through two distinct processes. In the first phase, the expression of the target antigen was dependent on the dose of inoculated virus, without specific protein synthesis. The target antigen seems to be produced by virus‐cell fusion. In the second phase, the expression of the target antigen was accompanied by synthesis of an early protein. In spite of the difference in their mode of expression, the first‐phase and the second‐phase target antigens were cross‐reactive in cytotoxicity inhibition assays.

Cowpox virus, CPR Cl strain, brought about a lower response than vaccinia virus, IHD‐J strain, in both sensitization of CTL and formation of CTL‐susceptible cells at both the first and the second phase. The cross‐reactive, but inefficient, recognition of anti‐vaccinia CTL for cowpox‐infected cells suggested a slight difference in the target antigens of the two viruses. Attempts to identify the target antigen were then made by comparing the polypeptide composition of vaccinia virus, cowpox virus, and their recombinants. SDS‐PAGE analysis of trypsinactivated viruses revealed 44K (cowpox)/45K (vaccinia) polypeptides which corresponded to the difference in target cell formation. Trypsinization of the viruses also increased the ability of the virus to induce the production of CTL‐susceptible target cells.
Requirement of Carbohydrates for Cell Fusion Induced by Vesicular Stomatitis Virus
Authors: Takehara Manabu
Published: November 2013

The fusion of BHK‐21‐KB cells by vesicular stomatitis virus was not induced in Eagle's minimal essential medium without glucose. In medium containing glucose, the rate of polykaryocyte formation decreased as the concentration of glucose was reduced below 5 mM. However, no reduction in virus production 24 hr after infection was seen under this condition. Addition of pyruvate or mannose to the culture medium caused a reversal of cell fusing activity. Cell fusion and virus growth were significantly suppressed by sodium azide and 2,4‐dinitrophenol.
Degradation of Ribosomal RNA in Bacteriophage Lambda Lysogens after Thermal Induction
Authors: Ono Tsuneko , Ohnishi Yoshinari
Published: November 2013

Stable RNA of Escherichia coli was extensively degraded about 40 min after thermal induction of lysogenized λcI857 phages at 42 C. When several nuclease‐deficient host cells were tested, RNase I activity in the host cells was inferred to be involved in the RNA degradation. Ribosomal structure was detectably altered before the degradation of ribosomal RNA was observed. 30S and 50S subunits began to sediment at 25–28S and 45–48S, respectively, still containing intact RNA. Nonpermissive host cells lysogenized with λcI857 susR produced progeny phages in normal burst size after thermal induction and then degraded stable RNA, though they were not lysed. In contrast cells lysogenized with λcI857 susS produced ten times more progeny phages under the same condition, but did not degrade stable RNA. These results indicate that the λS gene product, which acts as a positive effector of lysis, induced the degradation of stable RNA, presumably by a still uncharacterized effect on the cytoplasmic membrane.
Studies on the Termination of Immunological Tolerance in the Mouse Thymus Cell Population after Irradiation
Authors: Amagai Takashi
Published: November 2013

Immunological tolerance in the mouse thymus cell population induced by the intravenous injection of deaggregated bovine gamma globulin was terminated by whole body irradiation. After irradiation, the weight of the thymus recovered biphasically, and the termination of tolerance occurred as early as in the first phase. Both Thy‐1 antigen expression and helper activity of the thymus cell population in irradiated mice recovered in parallel with the recovery of the thymus weight. Sensitivity of the regenerating thymus cells to the tolerogen was not different from that of the normal thymus cells. The first phase of thymus regeneration may be caused by the proliferation and differentiation of relatively radioresistant and tolerogen insensitive precursors residing in the thymus. Tolerogen and/or immunogen reactive thymus cells may originate from the precursor.
Inhibitory Effect of 1‐β‐D‐Arabinofuranosylthymine on Synthesis of Epstein‐Barr Virus
Authors: Yonemura Kensuke , Sairenji Takeshi , Hinuma Yorio
Published: November 2013

The effect of 1‐β‐d‐arabinofuranosylthymine (ara‐T) on cell growth and synthesis of Epstein‐Barr virus (EBV) in human lymphoblastoid cell lines was determined. The growth of P3HR‐1 cells was not inhibited by 1 μg of the drug per ml; however, infectious virus production was strongly inhibited and was accompanied by decreased expression of early antigen (EA) and viral capsid antigen (VCA). The ability of 12‐O‐tetradecanoylphorbol‐13‐acetate or n‐butyric acid to induce synthesis of VCA, but not EA, in P3HR‐1 cells was inhibited by ara‐T. Similarly, VCA synthesis but not EA synthesis was inhibited by ara‐T in Jijoye cells superinfected with the P3HR‐1 strain of EBV. The results suggest that ara‐T has a specific inhibitory action against EBV replication.
Regulatory Mechanism of Delayed‐Type Hypersensitivity in Mice: III. In Vitro Analysis of Memory T Cells Involved in Augmentation of DTH Responses
Authors: Tamura Shin‐Ichi , Kojima Asato , Egashira Yasuyuki
Published: November 2013

The memory of delayed‐type hypersensitivity (DTH), manifested by the augmented responsiveness upon challenge with alum‐absorbed ovalbumin (OA), was induced in mice primed 7 days, 21 days, or 90 days previously with 1 μg of reduced and alkylated OA. The memory cells involved in the augmentation of DTH responses were analyzed in the in vitro induction system of T cells which mediate DTH against OA. Spleen cells from the primed mice generated DTH‐effector T cells (DTH‐Te) in a significantly accelerated fashion, compared with unprimed spleen cells, when cultured with OA. The accelerated generation of DTH‐Te in vitro was induced antigen specifically and was dependent on a certain T cell population in the primed spleen. The T cell population was found in the spleen of primed mice for at least 3 months after priming, corresponding to the persistence of DTH‐memory in vivo. Moreover, it was fractionated in the high‐density layer by discontinuous bovine serum albumin gradient centrifugation. The high‐density cell population decreased in density with increase in the time of culture and developed into DTH‐Te, which were separated in the low‐density layer on day 4 of culture. These results indicate that the T cells involved in the accelerated generation of DTH‐Te in vitro are long‐lived DTH‐memory T cells, which are probably precursor cells, capable of differentiating into DTH‐Te upon challenge with the antigen.
Restriction Endonuclease Analysis of the Genomes of Virulent and Avirulent Marek's Disease Viruses
Authors: Hirai Kanji , Ikuta Kazuyoshi , Kato Shiro
Published: November 2013

The DNAs from virulent strain BC‐1 and avirulent strain C2(A) of Marek's disease virus (MDV) were compared by electrophoresis on 0.5% agarose gels of the products obtained with the restriction endonucleases Bam H1, Sal 1, and Sma 1. The patterns of the fragments of the DNAs from these two strains were very similar, but showed some significant differences in the number and mobility of the DNA bands. The DNA fragments obtained with the restriction endonucleases, and especially Sma 1, were mostly present in equal molar ratios, but a few of those obtained with Bam H1 and Sal 1 were present in submolar amounts. In addition, several fragments obtained with Bam H1 and Sal 1 were present in greater than molar quantities, suggesting the presence of reiterated sequences in MDV DNA. The terminal fragments of MDV DNA were identified by their sensitivity to lambda 5′‐exonuclease. The terminal fragments obtained with Bam H1 A and Sal 1B showed heterogeneous electrophoretic mobility and contained sequences with high content of guanosine and cytosine, suggesting the presence of reiterated sequences at the end of the MDV DNA molecule.
Attachment of Mycoplasma hominis and M. orale to Human Diploid Lung Fibroblasts
Authors: Kihara Kojo , Yamada Kenichiro , Sasaki Tsuguo , Shintani Miharu , Okumura Hideo
Published: November 2013

The process of attachment of Mycoplasma hominis and M. orale to HAIN‐55 cells, derived from normal embryonic human lung, was investigated quantitatively. The attachment reached its maximum within about 2–4 hr at 37 C and increased linearly as a function of the number of organisms present in the system. The relative attachment efficiency of M. hominis was approximately 1% under our experimental conditions. Trypsin and EDTA were effective in detaching particles of M. hominis and M. orale from the surfaces of HAIN‐55 cells. Therefore it was suggested that some proteinaceous substance and salt bridges might be involved in the attachment of these mycoplasmas to HAIN‐55 cells.
Phenotypic Change of an SV40‐Transformed Mouse Macrophage Line, BB‐W‐531–2, Induced by Different Cultural Methods
Authors: Takayama Hisao , Tanigawa Takahiko , Takagi Atsushi
Published: November 2013

A simian virus 40 (SV40)‐transformed macrophage clone which was established from BALB/cAnN mouse bone marrow cells was used to study the effect of different cultural conditions on the expression of macrophage properties. The macrophage clone, BB‐W‐531–2 line, expressed and maintained the macrophage properties, immune phagocytosis or Fc‐ and complement receptors, under the growth‐inhibiting conditions of confluent density and of cultivation on bacteriologic dishes with reduced adhesiveness. However, the cells lost their ability to express the macrophage properties dependent upon cell density after repeated culture splits in the growing phase. These cells regained that ability when they were cocultured with cells having macrophage properties. These results suggest that there is a possible correlation between reduced multiplication and the expression of macrophage properties, and that macrophage properties which have been suppressed or blocked may be induced by diffusible factor(s) produced by macrophages.
Characterization of Two Inducible Bacteriophages, α1 and α2, Isolated from Clostridium botulinum Type A 190L and Their Deoxyribonucleic Acids
Authors: Kinouchi Takemi , Takumi Kenji , Kawata Tomio
Published: November 2013

Two inducible bacteriophages, α1 and α2, isolated from Clostridium botulinum type A strain 190L and their deoxyribonucleic acids (DNAs) were purified and characterized. Phage αl, which is unable to form plaques on any strain of C. botulinum, was produced in large quantities after treatment with mitomycin C (MC), whereas phage α2, which was induced in much lower quantities than phage αl, propagated in cultures of type A strain Hall. The phage DNAs were exclusively synthesized after induction with MC. αl and α2 DNAs had sedimentation coefficients of 34.0 and 30.6 S, corresponding to molecular weights of 31.9 × 106 and 23.5 × 106, respectively. The buoyant density in CsCl was 1.682 g/cm3 for al DNA and 1.680 g/cm3 for α2 DNA. Based on thermal denaturation characteristics, the genomes of both phages were shown to be double‐stranded DNAs. Agarose gel electrophoretic profiles of the phage DNAs digested with restriction endonuclease EcoRI revealed nine fragments for al DNA and six fragments for α2 DNA. The molecular weights of the phage DNAs as determined by restriction enzyme analysis were 30.55 × 106 for α1 DNA and 25.83 × 106 for α2 DNA. Nontoxigenic mutants obtained from strain 190L could, like the toxigenic parent strain, produce the two phages after treatment with MC. Lysogenic conversion to toxigenicity by phage α2 was not observed with the nontoxigenic mutants. It seems likely that there is no relationship between either phage genome and the toxigenicity of C. botulinum type A.
Adherence of Fimbriate and Non‐Fimbriate Strains of Yersinia enterocolitica to Human Epithelial Cells
Authors: Old David C. , Robertson Janet
Published: November 2013

Strains of Yersinia enterocolitica from different O serogroups were tested for their ability to adhere to cultured epithelial cells (H. Ep 2) after growth of bacteria in broth at 22 C or 37 C. Strains that adhered to H. Ep 2 cells generally did so better after growth at 22 C than at 37 C. The data indicate that the ability to produce either of the mannose‐resistant (MR) hemagglutinins, MR/Y or Kl, each associated with fimbriae, does not correlate with ability to adhere to epithelial cells in this particular in vitro model.
Relationship between Pigment Producibility and Drug Resistance in Serratia marcescens
Authors: Muto Yumiko , Tsuji Akiyoshi , Kaneko Yasuko , Goto Sachiko
Published: November 2013

Among the clinical isolates of Serratia marcescens, non‐pigmented cells appeared more frequently from pigmented, drug‐resistant strains than from pigmented, drug‐sensitive strains. Transfer of R plasmid from Escherichia coli to pigmented strains caused spontaneous loss of pigment producibility, whereas such spontaneous loss never occurred in fresh cultures of drug‐sensitive strains. The non‐pigmented strain was a better recipient of R plasmid from E. coli than was the pigmented strain. R plasmid was transferred from the non‐pigmented strain to the pigmented strain at a higher frequency than from E. coli to the pigmented strain. The results of the present investigation suggest that transfer of R plasmid may be one of the reasons for the significant increase of non‐pigmented, drug‐resistant strains of S. marcescens in nature.
Selective and Differential Media for Isolation and Tentative Identification of Each Species of Pityrosporum Residing on Normal or Diseased Human Skin
Authors: Ushijima Tsutomu , Takahashi Megumi , Ozaki Yoshikatsu
Published: November 2013

Selective and differential media were designed for each species of Pityrosporum; P. pachydermatis, P. ovale, and P. orbiculare in order to make feasible a quantitative cultivation.

Medium for P. pachydermatis (medium A) was composed of 1% trypticase peptone (BBL), 0.5% yeast extract (BBL), 0.3% glucose, 0.2% NaCl, 1.2% KH2 PO4 (anhydrous), 1.5% agar, 0.01% ampicillin, and 0.025% cycloheximide with a pH of 5.5. Medium for P. ovale (medium B) was medium A supplemented with 0.05% sodium acetate (anhydrous), 0.2% Tween 80, and 0.025% (selective medium) or 0.075% (differential medium) sodium laurate. Medium for P. orbiculare was medium B (devoid of laurate) supplemented with 2% olive oil, 0.25% glycerol, 0.25% gall powder, 0.05% sodium palmitate, 0.05% sodium stearate, 0.05% sodium oleate and 8% (selective medium) or 10% (differential medium) sodium lactate and an increase in Tween to 1%.

For isolation of Pityrosporum, specimens were suspended in 0.1% Tween 80 solution and inoculated onto agar plates of three selective media. The plates were incubated aerobically at 37 C for 8–10 days under conditions of prevention of water loss from the media. The plating efficiency of each selective medium, expressed as a ratio of cultural counts to microscopic counts was generally over 70%. Species of Pityrosporum could also be identified when we inoculated the cell suspension onto differential agar plates and incubated the preparations at 37 C for 7 days.
Characterization of Solubilized Products from the Cell Wall of Streptococcus pneumoniae during Autoplast Formation
Authors: Ohno Naohito , Yadomae Toshiro , Miyazaki Toshio
Published: November 2013

The structure of the cell wall of Streptococcus pneumoniae R36a was investigated by means of chemical analysis of the solubilized products formed during autoplast formation. By autoplast formation, almost all of the cell wall components were solubilized as the end products within several hours. Analysis of the solubilized products revealed that the pneumococcal cell wall consists of three macromolecular components, teichoic acid‐glycopeptide I (TA‐GP I), teichoic acid‐glycopeptide II (TA‐GP II), and glycopeptide (GP III). The molecular size of TA‐GP I was larger than that of TA‐GP II. TA‐GP I and TA‐GP II were constituted of similar components, galactosamine, 2‐acetamido‐4‐amino‐2, 4, 6‐trideoxyhexose, glucose, ribitol, choline, phosphate, and peptidoglycan components, but the ratio of teichoic acid to glycopeptide in TA‐GP II was higher than that in TA‐GP I. TA‐GP II was solubilized more slowly than TA‐GP I and GP III during autoplast formation. The assembly of the cell wall by TA‐GP I and II, and GP III is discussed in connection with the action of autolysin.
Further Studies of the Polysaccharide of Klebsiella pneumoniae Possessing Strong Adjuvanticity: II. Serological Relationship between the Adjuvant Polysaccharide and O3 Antigen of Klebsiella
Authors: Kato Nobuo , Ohta Michio , Hasegawa Takaaki , Mori Masashi , Yamaki Kenichi , Mizuta Kazutaka , Nakashima Izumi , Naito Setsuko
Published: November 2013

The serological specificity of the neutral polysaccharide possessing extraordinarily strong adjuvanticity originally isolated from the culture supernatant of Klebsiella K1 strain Kasuya has been investigated. Among all of the reference strains (K1–K82) of Klebsiella obtained from the International Escherichia and Klebsiella Center, Statens Seruminstitut, Copenhagen, only 13 strains have been shown to produce the adjuvant polysaccharide by the passive hemagglutination inhibition test. All of these 13 strains belong to the 03 group, and the strains which belong to other O groups or O groups of which were not identifiable did not produce it. The gel precipitation test has demonstrated that the adjuvant polysaccharide is antigenically identical to O3 antigen isolated from the cells of the decapsulated mutant (strain LEN 1) of Klebsiella K1 strain Kasuya and to O9 antigen of Escherichia coli isolated from either the culture supernatant or the cells, which has already been shown to be antigenically and structurally identical to the O3 antigen of Klebsiella.
Cell Surface Expression of I‐A Products Is Required for Contact Sensitivity Induction by Trinitrophenyl‐Coupled Epidermal Cells
Authors: Ikezawa Zenro , Sato Masahiro , Nagai Ryukichi , Okuda Kenji
Published: November 2013

Trinitrophenyl (TNP)‐coupled epidermal cells (EC) injected subcutaneously (s.c.) were more capable of inducing contact sensitivity (CS) to 2, 4, 6‐trinitro‐1‐chlorobenzene (TNCB) than similarly substituted spleen cells (TNP‐SC). Furthermore, the intravenous (i.v.) or intraperitoneal (i.p.) injection of TNP‐EC also induced CS responses, whereas the i.v. or i.p. injection of TNP‐SC failed to induce them. Treatment of mice with cyclophosphamide (Cy; 50 mg/kg) or anti I‐J serum allowed animals injected with TNP‐SC i.v. to develop significant CS responses, suggesting that Cy‐sensitive and I‐J positive regulatory cells were involved in the induction of unresponsiveness by the i.v. injection of TNP‐SC. Mapping studies of the major histocompatibility gene complex (MHC) region demonstrated that identity at the I‐A subregion alone between EC donor and recipient mice was sufficient for the induction of CS by TNP‐EC given i.v. Blocking experiments using antisera in the absence of complement indicated that I‐A subregion‐encoded antigens on the surface of TNP‐EC apparently are involved in the induction of CS, and are not simply phenotypic markers on the surface of accessory cells.