bluuebunny/crossref_metadata_2025 huggingface dataset:
Fluctuation of Antigen Binding Activity during the Cell Cycle in the Synchronized Population of the Murine T Hybridoma Cell Line
Authors: Kuwata Shigeki , Namba Yuziro , Hanaoka Masao
Published: November 2013

The murine T cell hybridoma line which specifically binds antigen (ovalbumin) was established using a cell fusion technique with Sendai virus. Regional lymph node cells from ovalbumin (OVA) immunized C57BL/6 mice were fused with thymidine kinase deficient variant cells of the EL‐4 cell line (originating from a thymoma of a C57BL/6 mouse). Approximately one hundred cell lines were established and the antigen binding activity was determined by rosette formation with OVA coated sheep red blood cells (SRBC). One hybridoma cell line, MMH‐77, could form rosettes and this formation was specifically inhibited by the addition of free OVA. The ability of the cell line to form rosettes varied from one stage of the cell cycle to the other with the maximum ability in the S phase.
Study of the Mode of Action of Ribosomal Vaccines from Klebsiella and Streptococcus pneumoniae and Their Ribonucleic and Protein Fractions Using Passive Immunization
Authors: Robert Daniel , Ivanoff Bernard , Fontanges Robert , Normier Gérard , Pinel Anne Marie , D'Hinterland Lucien Dussourd
Published: November 2013

The vaccinating potency of ribosomal fractions of Klebsiella pneumoniae and Streptococcus pneumoniae as well as their ribosomal RNA and protein fractions has been studied with respect to their ability to induce cellular or humoral immunity. Experiments with transfer of serum or spleen cells from vaccinated animals have shown that anti‐Klebsiella immunity is essentially cellular, while streptococcal immunity is exclusively humoral. Results have been discussed as a function of differential results for the various fractions under study.
An Attempt to Localize the Vaccinating Power of Klebsiella pneumoniae Ribosomal Preparations Using Saccharose‐Gradient Ultracentrifugation
Authors: Robert Daniel , Bienvenu Pierre , Lafont Sergyl , Jouanneteau Bernard , Normier Gerard , D'Hinterland Lucien Dussourd , Fontanges Robert
Published: November 2013

The study of the vaccinating power of ribosomal vaccines against Klebsiella pneumoniae led us to define the chemical nature which supports this protective activity. We tried to separate this support and the ribosomes by sucrose gradient ultracentrifugation. We isolated high protective membrane vesicles by this technique applied to salt‐washed ribosomal preparations. When the ribosomal preparations were exposed to SDS, the protective activity was conserved all along the gradient, with no correlation with the ribosome concentration. The addition of bovine serum albumin to the ribosomal preparation focused the protective activity on the ribosomal peak. No correlation was observed between the response to capsular polysaccharide and the vaccinating power of the fractions.
Adhesion of Haemophilus paragallinarum to Cultured Chicken Cells
Authors: Ueda Susumu , Nagasawa Yoji , Suzuki Taeko , Tajima Masanori
Published: November 2013

Adhesion of Haemophilus paragallinarum to chicken embryo fibroblasts (CEF) in vitro was investigated in correlation with its virulence. Under the scanning electron microscope, the organisms were seen adhering to CEF exposed to the bacteria. By transmission electron microscopy, the organisms appeared to attach to the plasma membrane of CEF by their fuzzy material. They were enclosed in membrane‐limited vesicles and appeared morphologically intact. Specific fluorescence was seen in the cytoplasm of CEF in later stages of infection. There was a good correlation between pathogenicity of the organisms for chickens and their ability to adhere to CEF. This correlation was reinforced by the fact that the ability of a pathogenic strain to adhere to the mucosal surface of the chicken trachea was demonstrated by scanning electron microscopy.
Metabolic Disorders of Serum Lipoproteins in Endotoxin‐Poisoned Mice: The Role of High Density Lipoprotein (HDL) and Triglyceride‐Rich Lipoproteins
Authors: Sakaguchi Shuhei
Published: November 2013

A study was performed to clarify the role of serum lipoproteins, especially high density lipoprotein (HDL) and triglyceride‐rich lipoproteins in endotoxemic or endotoxin‐poisoned animals. The level of HDL‐cholesterol decreased markedly in mouse serum 18–24 hr postintoxication, while the amount of low density lipoprotein (LDL)‐cholesterol in the sera of poisoned mice was about 175% of that of the controls. Serum lecithin‐cholesterol acyltransferase activity in the poisoned mice decreased slightly for 3–6 hr after endotoxin injection, but became markedly increased at 18–24 hr as compared with that in the controls. The amount of serum very low density lipoprotein (VLDL) showed a marked increase in the poisoned mice 8–24 hr postintoxication. The HDL fraction in the electrophoretic patterns of serum was reduced according to the dose of endotoxin 18 hr postintoxication. The HDL fraction in mice injected with lead acetate plus endotoxin was markedly lower than that in the poisoned mice. When streptozotocin‐diabetic mice were injected with endotoxin, the HDL fraction was higher than that in the endotoxin‐poisoned mice. In endotoxin‐poisoned mice a correlation was observed between the lipid peroxide and LDL levels in the serum. In disk electrophoretic patterns, the HDL fraction in mice given vitamin E‐supplemented diet showed a higher level than that in mice given a normal diet. Lipoprotein lipase (LPL) activity in poisoned mice significantly decreased to 59% of the control value 18 hr postintoxication, but hepatic triglyceride lipase activity was only slightly increased in endotoxin‐poisoned mice. In analysis of HDL apoprotein peptide in serum lipoprotein, the apo C‐II peptide level was clearly lower in mouse serum 18 hr postintoxication than that in the controls.

These results suggest that the decrease in LPL activity in endotoxin‐poisoned mice may be closely related to a decrease in the apo C‐II peptide level, and also that it plays an important part in HDL and triglyceride‐rich lipoprotein metabolism in the poisoned mice.
Restricted Viral RNA Synthesis in Establishment of Persistent Infection in Vero Cells with a Sendei Virus Mutant
Authors: Kanda Tadahito , Shibuta Hiroshi
Published: November 2013

It was previously shown that a temperature‐sensitive mutant of Sendai virus, ts‐23, readily establishes persistent infection in Vero cells at 37 C, a permissive temperature for growth of the mutant.

In the present study, it was demonstrated that the virus yield from ts‐23‐infected Vero cells at 37 C began to decrease 48 to 72 hr postinfection, after an initial phase of high virus production. Before the decrease in virus production, the formation of viral nucleoprotein declined, although synthesis of all species of viral protein continued. It was suggested that the limited formation of viral nucleoprotein and the decrease in virus production were due to the restriction of viral RNA synthesis which began to occur early after infection in ts‐23‐infected cells at 37 C. The mutant has a temperature‐sensitive defect in RNA polymerase activity and the temperature 37 C, used for establishment of persistent infection, would be a semi‐permissive temperature for the RNA polymerase activity of the mutant. The ts‐23 mutant interfered with the replication of the parental wild virus in Vero cells at 37 C.
Occurrence of Uronic Acid in Lipopolysaccharides of Vibrionaceae
Authors: Hisatsune Kazuhito , Kondo Seiichi , Iguchi Takehiro , Machida Masaaki , Asou Shinobu , Inaguma Makoto , Yamamoto Fumihiro
Published: November 2013

The occurrence of uronic acid as a sugar constituent of lipopolysaccharides (LPS) in Vibrionaceae was demonstrated for the first time. More than 100 strains were examined. Of five genera constituting Vibrionaceae, i.e., Vibrio, Aeromonas, Plesiomonas, Photobacterium, and Lucibacterium, the latter three contained uronic acid in LPS of all of their constituting members examined, while it was totally lacking in Aeromonas LPS so far tested. Only the members of genus Vibrio were found to be divided into uronic acid‐containing and ‐lacking groups; V. parahaemolyticus, V. alginolyticus, V. fisheri, V. costicola, Beneckea (‘Vibrio’), and V. fluvialis belonged to the former, while all four biotypes of V. cholerae regardless of their serotypes, V. vulnificus and V. anguillarum, belonged to the latter group.

The uronic acid content of V. parahaemolyticus O1 to O12 LPS ranged from 1.6 to 4.2%. The uronic acid residue released from V. parahaemolyticus O1, O4, O10, and O12 LPS by heating in 5% acetic acid at 100 C for 2 hr was identified as galacturonic acid; in particular, that from 012 LPS was characterized as d‐galacturonic acid.
Effector Cells Working for Rejection of Ad12 Tumors in Mice
Authors: Maeta Yasunobu , Toshima Muneatsu , Miura Rinjiro , Yamai Tereza Y. , Hamada Chuya
Published: November 2013

Immune spleen cells (ISC) capable of inhibiting the growth of adenovirus type 12 (Ad12) tumors were raised in C57BL/6 mice by immunization with Ad12, fractionated according to their affinity for plastic and nylon‐wool substrates or treated with various antisera plus complement, and subjected to the tumor‐neutralization test (Winn) to define the effector cells for the cell species. Full antitumor activity of ISC was recovered in the cell fractions nonadherent to the two substrates; the antitumor activity of ISC was abrogated entirely by anti‐Thy‐1,2 serum and almost entirely by anti‐Lyt‐2.2 ascites fluid plus complement. These results clearly indicate that T‐lymphocytes, particularly those bearing Lyt‐2.2 antigen, are the principal effectors in ISC against Ad12 tumors in animals.
Protective Effects of Antibody against Intestinal Invasion by Escherichia coli
Authors: Okabe Tatsuji
Published: November 2013

Seven Escherichia coli isolates from newborn calves with diarrhea were examined for enteropathogenic properties. One isolate penetrated into HeLa cells, four produced enterotoxin(s) and the remaining two possessed neither of these properties. Penetration of E. coli into HeLa cells was inhibited by antibody in bovine colostrum and in bovine and rabbit immune sera. The effective antibodies appeared to be mostly of the IgM class. The invasion by E. coli isolates was also examined by inoculation of the bacteria into the small intestine of E. coli‐immunized and non‐immunized guinea pigs. The isolate which penetrated into HeLa cells could penetrate the intestinal mucosa to be disseminated into various organs of non‐immunized guinea pigs but not of immunized guinea pigs, whereas no other isolates showed such pathogenicity in vivo. The inhibition of the invasion was observed when non‐immunized guinea pigs were inoculated with the bacteria together with colostral or serum antibody. The results show the importance of antibody in the local defense mechanism against E. coli invasion.
Variation in Virulence of Coxsackie Virus B3 in the Heart of Mice
Authors: Hashimoto Isao , Komatsu Toshihiko , Kohara Teruko
Published: November 2013

Histopathological analysis of the heart in adult mice inoculated with Coxsackie virus B3 (CB3) strains revealed that strain SK‐74 isolated from a patient suffering from severe diarrhea and fever produced severe myocarditis but strain T‐70 isolated from a healthy child induced no lesion in the hearts of mice tested, and that intensities of myocardial lesions in mice inoculated with strain PMH were higher than those in mice inoculated with prototype strain Nancy. The results support the conclusion in the preceding paper that strain SK‐74 is virulent but strain T‐70 is avirulent in mice. The results also partially indicated that the virulence of prototype strain Nancy in the heart of mice is enhanced by passages of the strain in the heart of mice. All four strains of CB3 produced lesions in the pancreas although lesions induced by strain T‐70 were less marked than those induced by the remaining three strains.
Epidemiological Studies on Echovirus Type 18 Infection in Toyama Prefecture
Authors: Matsuura Kumiko , Hasegawa Sumiyo , Nakayama Takashi , Morita Osayuki , Uetake Hisao , Masaki Akio
Published: November 2013

Echovirus type 18 (echo 18) was isolated from six aseptic meningitis children in Fukumitsu‐machi, Toyama Prefecture, from July to August, 1980. This was the first virologically‐confirmed epidemic of aseptic meningitis due to echo 18 in Japan. Epidemiological studies on the prevalence of this virus among the inhabitants in Toyama Prefecture were also performed. The results are summarized as follows. (1) Significant increases in neutralizing antibody titers against echo 18 were observed in all the paired sera of aseptic meningitis patients from whom echo 18 was isolated. (2) In October, 1980, echo 18 was also isolated from healthy children or from infants suffering from gastroenteritis in other areas of Toyama Prefecture. (3) Among the sera collected from 50 children (aged ≤ 12) in Fukumitsumachi in December, 1980, neutralizing antibodies against echo 18 were detected only in the younger groups (aged ≤8), 58.9% of these age groups showing a titer of 4 or higher. (4) Another epidemic around 1963 by echo 18 in Toyama Prefecture was retrospectively suggested by the examinations of sera collected in 1978 and 1981 from inhabitants in various areas of Toyama Prefecture. (5) Neutralizing antibody titers against the strain isolated, No. 35′80, were significantly higher than those against strain ‘Metcalf,’ the prototype of echo 18, in most sera including both aseptic meningitis patients and healthy inhabitants.
Interaction of the Eighth Component of Guinea Pig Complement (C8) with the Membrane‐Bound C5b‐7 Complex
Authors: Kinoshita Taroh , Hong Kyongsu , Inoue Kozo
Published: November 2013

The eighth component (C8) of guinea pig complement consists of three polypeptide chains, the α‐, β‐, and γ‐chains with M.W. of 60,000, 60,000, and 24,000, respectively. The α‐ and γ‐chains are bound by a disulfide bond(s) forming an α‐γ subunit, which is linked noncovalently to the β‐chain.

The α‐γ subunit and the β‐chain were separated and purified from C8 by treatment with sodium dodecyl sulfate (SDS) and gel chromatography on Sephacryl S‐300 in the presence of SDS.

After removal of SDS, neither α‐γ nor β showed the hemolytic activity of C8 when assayed independently, but showed significant activity in combination, indicating reconstitution of active C8. The recovery of hemolytic activity was 3.48%. When α‐γ and β were incubated successively with EAC‐7 with intervening washing, reconstitution of active C8 on the cells was insignificant, irrespective of the order of the reactions. α‐γ and β did not bind to EAC‐7 when added separately, but after recombination 7% of α‐γ and 9% of β bound to EAC‐7 when EAC‐7 was in excess.

These results indicate that the binding site of guinea pig C8 to the membrane‐bound C5b‐7 complex does not exist on either α‐γ or β only but stretches over both or is formed on one subunit after recombination of the subunits.
Mechanism of Antibiotic Resistance in Mycobacterium intracellulare
Authors: Mizuguchi Yasuo , Udou Takezo , Yamada Takeshi
Published: November 2013

The mechanism of resistance of Mycobacterium intracellulare strain 103 and other clinical isolates to a variety of drugs including aminoglycoside and peptide antibiotics was investigated. Enzymatic inactivation of aminoglycoside and peptide antibiotics could not be demonstrated. Ribosomes of the strain were found to be sensitive to the antibiotics. The levels of resistance of strain 103 and other clinical isolates decreased dramatically when the culture medium was changed from Dubos agar to Tween 80‐containing agar. These results suggest that a permeability barrier is the reason for naturally occurring resistance in M. intracellulare.
Conjugal Transferability of Multiple Resistance in Shigella Strains
Authors: Tanaka Tokumitsu , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

Shigella strains isolated in Japan between 1971 and 1979 were surveyed for drug resistance and distribution of R plasmids. Of 2,510 strains, 89.3% were resistant to either one or various combinations of four drugs, tetracycline, chloramphenicol, streptomycin, and sulfanilamide. About 66% of the Shigella isolates were quadruply resistant. The frequency of isolation of R plasmids from quadruply resistant Shigella strains was the highest when compared with other strains resistant to various combinations of the four drugs. The conjugal transferability of 204 quadruply resistant strains isolated between 1977 and 1979 was tested by various mixed‐culture methods. Among the total strains examined, 70.6% carried transferable resistance when tested by the conventional broth culture method, 90.2% transferred their resistance when, in addition the replica‐plating methcd was used and 97.5% could transfer their resistance when the membrane filter method was also used. Although the remaining five strains could not transfer their resistance by any of the mixed culture methods, the drug resistance of four of the five strains was mobilized by the concomitant presence of F‐tet or T‐kan plasmid.

These results indicate that almost all of the quadruple resistance in Shigella isolates was mediated by plasmid.
Subcellular Location of the Major Protein Antigens of Paramyxoviruses Revealed by Immunoperoxidase Cytochemistry
Authors: Nagai Yoshiyuki , Yoshida Tetsuya , Hamaguchi Michinari , Nagura Hiroshi , Hasegawa Hideki , Yoshimura Shinichi , Watanabe Keiichi
Published: November 2013

The major structural proteins of Newcastle disease virus and Sendai virus were localized in infected BHK‐21 and MDBK cells by ultrastructural immunoperoxidase cytochemistry using antibodies against the individual viral protein antigens. The intracellular glycoproteins were strictly membrane bound, being localized in the rough endoplasmic reticulum (RER), perinuclear spaces, smooth membrane vesicles, and presumed Golgi apparatus. The nucleocapsid proteins were detected exclusively in membrane free cytosol and accumulated there, forming inclusions. The membrane (M) protein was found both in cytosol and on RER. The viral proteins on RER exhibited a distinct site specificity; the glycoproteins were facing the lumen of RER whereas M protein was present at the outer cytoplasmic surface. All the viral proteins were detectable at the plasma membrane where virus assembly takes place. However, their modes of distribution differed remarkably. The glycoproteins were spread widely over the entire cell surface including the areas of virus budding and those of normal morphology, whereas M protein was localized in restricted areas of the membrane, frequently forming a patch of virus specific membrane. The presence of nucleocapsids was confined to the virus particles budding from the plasma membrane. These results complement and extend the earlier morphological and biochemical data on the assembly or morphogenesis of paramyxoviruses.
Antiviral and Antiproliferative Activities of Three Molecular Components of Leukocyte‐Derived Human Interferon
Authors: Dinolfo Elaine , Chadha Kailash C.
Published: November 2013

Antiviral and antiproliferative activities of three naturally occurring components of leukocyte‐derived human interferon (HuIFN‐α) separated by concanavalin A‐agarose affinity chromatography were studied in a variety of neoplastic cell lines. Significant differences were seen with these different components. The results strongly suggest that careful consideration must be given while selecting any one component of HuIFN‐α for any of the antiviral or antiproliferative studies. There is no clear evidence that glycosylation of HuIFN‐α has any significant influence on its in vitro antiviral or antiproliferative activities, although apparently glycosylated and non‐glycosylated components gave different antiviral and antiproliferative responses in different tumor cells.
Absence of Detectable Cytotoxic Lymphocytes in the Spleens of Guinea Pigs Immunized with Allogeneic Cells
Authors: Kataoka Tetsuro , Tokunaga Tohru
Published: November 2013

Using inbred guinea pigs of strains JY 1, JY 2, 2, and 13, whose major histocompatibility gene complexes are different from each other at the B region and/or the I region, the authors aimed to detect cytotoxic thymus‐derived lymphocytes (CTL) against allogeneic target cells. Various in vivo and in vitro sensitization methods that have been described for murine CTL systems were used. For detection of CTL, Con A‐ or PHA‐induced 51Cr‐labeled lymphoblasts were incubated with sensitized spleen cells or lymph node cells for 4 hr, and 51Cr released into the supernatant was assayed. Under these conditions, no significant lysis of the target cells was observed. On the other hand, significant levels of both delayed‐type hypersensitivity and humoral antibodies were detected in the animals sensitized by in vivo methods.
Morphological Differences in Flagella in Campylobacter fetus Subsp. intestinalis and C. fetus Subsp. jejuni
Authors: Morooka Tatsuya , Umeda Akiko , Amako Kazunobu
Published: November 2013

Intact flagella were isolated from human pathogenic strains of Campylobacter, C. fetus subsp. intestinalis and C. fetus subsp. jejuni, by the method of DePamphilis and Adler and examined by electron microscopy. The isolated flagella were composed of a filament, a hook, a basal body, and a large disk associated with the end of the hook region covering the basal body. The width of the hook was approximately 28 nm, somewhat greater than that of the filament (20 nm in diameter). The hook region of C. fetus subsp. intestinalis was curved, but it was straight in C. fetus subsp. jejuni. The structure of the basal body of the two subspecies was similar to that reported for other gram‐negative bacteria. The large disk detached from the flagella showed concentrically arranged circular structures. This structure was more clearly observed in the disk of C. fetus subsp. jejuni than in C. fetus subsp. intestinalis. Observations of thin‐sectioned profiles at the attachment site of the flagellum revealed that the large disk is located on the inner side of the outer membrane. The role of the large disk in bacterial movement is not clear, but it is assumed that it acts as an organ to protect the flagellar insertion site from vigorous rotation of the polar end inflicted during bacterial movement.
Elicitation of Delayed Type Hypersensitivity in Chicks after In Ovo Sensitization with Different Molecular Forms of the Same Hapten
Authors: Moriya Osamu , Ichikawa Yoichi
Published: November 2013

Lymphocyte sensitization, which participates in delayed type hypersensitivity (DTH) in chick embryos, was studied. The in ovo injection of dinitrophenylated keyhole limpet hemocyanine (DNP‐KLH) or DNP‐dextran (DNP‐D) led to delayed onset of the hapten‐specific reaction as shown by allergic contact dermatitis (ACD) after hatching. The extent of the ACD response was not directly dependent on the antigen dosage or the number of injections given for sensitizing. The magnitude of the ACD response was higher in chicks sensitized with DNP‐D than in those given DNP‐KLH. These findings suggest the presence of embryonic lymphocytes which can be sensitized by in ovo antigenic stimulation at the later stage of embryogenesis and may make possible the differentiation of functional lymphocytes. Antigen stimulation with higher doses may be inadequate for the in ovo sensitization of embryonic lymphocytes. The ACD response elicited by DNFB in chicks primed with either DNP‐KLH or DNP‐D was thought to be T‐cell dependent, from the kinetics of the ACD peak within a period of 24 to 48 hr. Furthermore the conditions for in ovo sensitization of embryonic lymphocytes by DNP‐D seem to be different from those for sensitization by DNP‐KLH.
Effective Immunity to Dental Caries: Protection of Gnotobiotic Rats by Local, Immunization with a Ribosomal Preparation from Streptococcus mutans
Authors: Gregory Richard L. , Michalek Suzanne M. , Shechmeister Isaac L. , McGhee Jerry R.
Published: November 2013

A ribosomal preparation from Streptococcus mutans 6715 was characterized for its ability to induce an immune response in gnotobiotic rats which was protective against S. mutans‐induced dental caries. Animals injected in the salivary gland region with the S. mutans ribosomal vaccine developed significantly higher (P < 0.01) salivary IgA and serum IgG antibody activities against whole S. mutans cells and ribosomal preparations than nonimmunized rats. Vaccinated animals had significantly lower (67%; P < 0.01) levels of S. mutans adherent to their molar surfaces than the control rats after infection with the homologous, cariogenic S. mutans. The immunized animals had significantly fewer (P < 0.01) carious lesions on their buccal, sulcal, and proximal molar surfaces than the nonimmunized rats following challenge with the virulent organism. Animals injected with the ribosomal preparation developed salivary IgA and serum IgG antibodies with specificities to various cell surface‐associated antigens such as lipoteichoic acid and glucosyltransferase, suggesting that the observed protection may be due to antibodies against cell surface contaminants of the ribosomal vaccine. These results are the first demonstration that a ribosomal preparation from S. mutans protected rats from caries formation after challenge with the homologous, virulent S. mutans.
Isolation and Partial Properties of a Porin‐Like Protein from Vibrio parahaemolyticus Cell Envelope
Authors: Koga Tetsuro , Kawata Tomio
Published: November 2013

The cell envelope of Vibrio parahaemolyticus pilot strain K‐11 contains a major protein with an apparent molecular weight of 35,000 which was not solubilized with 2% sodium dodecyl sulfate (SDS) at 50 C for 30 min and was resistant to trypsin. The protein was extracted from the SDS‐insoluble envelope with SDS containing 0.4 m NaCl and purified by acetone precipitation and gel filtration. The purified protein was completely dissociated into a monomer with a molecular weight of 35,000 in SDS at 60 C. The amino acid composition of the protein was nearly the same as that of porins from Escherichia coli and Salmonella typhimurium. Thus the protein seems to be porin‐like.
Type‐Specific and Cross‐Reactive Antigenicity of Capsid Proteins VP1 and VP2 of Echovirus Type 7
Authors: Hasegawa Ayako , Inouye Sakae
Published: November 2013

After disruption of echovirus type 7 virions with urea and heat, VP1 and VP2 were separated by isoelectric focusing in urea‐containing sucrose gradients. Antisera to these two polypeptides were produced in guinea pigs. In complement fixation, antiserum to VP1 reacted with native and heated virions (N and H antigens, respectively) of homologous virus, and also cross‐reacted with healed virions of some other enteroviruses used. Antiserum to VP2 was reactive only with heated virions of homologous and heterologous viruses. Interestingly, the anti‐VP2 serum reacted neither with native nor even with heated procapsids (naturally‐occurring empty capsids). Antiserum to VP1, but not VP2, showed neutralizing and hemagglutination‐inhibiting activities. These results suggest that 1) both VP1 and VP2 possess cross‐reactive antigenic determinants which are exposed on the surface of heated virions, and 2) type‐specific determinants of VP1 are located on the surface of native virions.
FcεR and FcγR Expressed on Murine IgE‐Specific Suppressor T Hybridomas
Authors: Yodoi Junji , Suemura Masaki , Kishimoto Tadamitsu
Published: November 2013

The expression of Fc receptors specific for IgE (FcεR) and those for IgG (FcγR) on murine IgE‐specific suppressor T hybridomas was studied. While parental T lymphoma cells (BW5147) failed to bind IgE‐sensitized red cells (mIgE‐TNP‐ORBC), the majority of T hybridoma cell lines having IgE‐specific suppressor activity contained rosette‐forming cells (RFC) binding mIgE‐TNP‐ORBC (2 to 13% of the total cells). The expression of FcεR was poor (2% or less) in T hybridoma cell lines without IgE‐specific suppressor activity. In addition to FcεR, IgE‐specific suppressor T hybridomas also expressed FcγR as detected by ORBC sensitized with IgG antibodies (EAoxγ). β‐Interferon (IFN) (1,000 to 2,000 U/ml) augmented the expression of FcγR, but not of FcεR. On the other hand, preculture of the cells with 40 μg of mIgE per ml enhanced the expression of FcεR without augmenting FcγR expression. IgE‐specific suppressor activity in the culture supernatants of T hybridomas was also augmented by preculture with mIgE. However, a dissociation between the expression of FcεR and IgE‐suppressor activity was observed. In some of the subclones, mIgE augmented the suppressor activity without inducing the expression of FcεR. Furthermore, when treated with 5 μg of melittin, a phospholipase A2 activator, per ml, IgE‐specific suppressor activity of the hybridomas was completely abrogated, whereas IgE‐induced expression of FcεR was enhanced by melittin.
A Clustering Outbreak of Hand, Foot, and Mouth Disease Caused by Coxsackie Virus A10
Authors: Itagaki Asao , Ishihara Junko , Mochida Kyo , Ito Yoshihiro , Saito Koichi , Nishino Yasuo , Koike Shigeyuki , Kurimura Takashi
Published: November 2013

A clustering outbreak of hand, foot, and mouth disease (HFMD) occurred from July, 1981 to January, 1982 in Matsue City and Gotsu City, Shimane Prefecture. Thirty‐seven patients with clinical HFMD were virologically and serologically examined, and Coxsackie virus A10 (CA10) was isolated in 18 patients from vesicles (7/16), throat‐swabs (9/31) and feces (6/7). During the period, no CA16 or enterovirus 71 were isolated from HFMD patients or from other diseases such as pharyngitis, febrile diseases, and aseptic meningitis. Serological diagnosis was performed employing an African green monkey kidney cell (AG‐1)‐adapted CA10 which demonstrated cytopathogenic effects on the cells. Paired sera from seven patients including three cases in which isolation failed showed a significant increase of neutralizing antibody titer against CA10. Finally, an etiological diagnosis was made in 21 out of 37 patients with clinical HFMD. This is the first report of a clustering outbreak of HFMD caused by CA10 in Japan.
Mouse Alloantibodies Capable of Blocking Cytotoxic T Cell Function
Authors: Kojima Masaharu , Taniguchi Masaru , Shinohara Nobukata
Published: November 2013

Blocking of cell‐mediated lympholysis (CML) by anti‐Lyt‐2 antibodies was compared with that by anti‐H‐2 antibodies which most likely inhibit CML by blocking antigen recognition by cytotoxic T lymphocytes (CTLs). Both antibodies were shown to inhibit the early Mg2+‐dependent process of killing. Moreover, the anti‐H‐2‐sensitive event was found to be reversible by the antibody as was the case with the anti‐Lyt‐2‐sensitive event, suggesting that the two antibodies block the same event taking place during the Mg2+‐dependent stage. Both types of antibody were also shown to be capable of inhibiting the phytohemagglutinin (PHA)‐mediated nonspecific killing activity of CTLs. However, in the case of anti‐Lyt‐2 antibodies, available monoclonal antibodies failed to inhibit PHA‐mediated killing whereas conventional antisera did. The results thus suggest multiplicity and heterogeneity of Lyt‐2 determinants or the existence of multiple products of Lyt‐2‐linked genes. In addition, an anti‐H‐2 antiserum also exerted a specific inhibitory effect on PHA‐mediated killing. Thus there appears to be a general requirement for involvement of the Lyt‐2 molecules on CTLs and major histocompatibility complex (MHC) products on the target cells. The implications of these observations are discussed.
T Cell‐Activating Monokines in Guinea Pigs: Comparison of High and Low Molecular Weight Factors
Authors: Sakamoto Shigeru , Onoue Kaoru , Ohishi Masamichi
Published: November 2013

Guinea pig monokines produced by lipopolysaccharide‐stimulated peritoneal macrophages were found in high (50,000–80,000) and low (10,000–30,000) molecular weight (m.w.) fractions by gel filtration. Both showed enhancing activity on the proliferative response of guinea pig and mouse thymocytes to PHA, but the high m.w. (65K) monokine was much more efficient than the low m.w. (15K) monokine in enhancing the response of lymph node T cells to PHA, suggesting its importance in the activation of peripheral T cells. The 65K monokine was coeluted with BSA present in the culture medium by DEAE‐cellulose chromatography, but was clearly separated from it by hydroxylapatite chromatography. The immunoadsorption experiment with anti‐BSA‐coupled gel also indicated that 65K monokine is not a complex of low m.w. monokine with BSA. Our series of studies showed that most monokine activities were always found in the 65K fraction in guinea pigs. Thus, in guinea pigs, the 65K component appears to constitute a major class of T cell‐activating moonkines.
Carbenicillin‐Hydrolyzing Penicillinases of Proteus mirabilis and the PSE‐Type Penicillinases of Pseudomonas aeruginosa
Authors: Takahashi Ikuko , Tsukamoto Kikuo , Harada Masako , Sawai Tetsuo
Published: November 2013

Three carbenicillin‐hydrolyzing penicillinases were found in Proteus mirabilis strains, N‐3, N‐29, and GN79. The former two strains were isolated in 1978, but strain GN79 was one of our stock cultures isolated in 1965. These penicillinases closely resembled each other, and the PSE‐1 and PSE‐4 enzymes produced by Pseudomonas aeruginosa, in their substrate profiles and kinetic properties for hydrolyzing various β‐lactams. However, differences were found in their molecular weights and isoelectric points which ranged from 22,000 to 27,000 and from 6.0 to 6.9, respectively. The antiserum against the purified penicillinase of N‐29 cross‐reacted with the enzyme of N‐3 and inhibited its activity by more than 80%. The antiserum also reacted with the PSE‐1 and PSE‐4 enzymes. The antiserum did not react with the penicillinase from strain GN79 and the PSE‐2 and PSE‐3 enzymes of P. aeruginosa. Enzyme production in N‐3 and N‐29 was mediated by R plasmids.
Sporulation Promoting Factor in Vegetative Cells of Bacillus subtilis
Authors: Hachisuka Yoetsu , Imura Hiromi , Nakata Akemi , Hattori Emiko
Published: November 2013

A simple experimental system for detection of sporulation promoting factors was presented. This system showed that there was a sporulation promoting factor in the vegetative cells of Bacillus subtilis cultivated on nutrient agar for 9 hr (at stage T0). The factor was partially purified from the sonicate of vegetative cells by ethanol fractionation, gel filtration, chromatography and preparative gel electrophoresis, and it was identified as manganese‐containing protein.
Plasmids in Vibrio parahemolyticus Strains Isolated in Japan and Bangladesh with Special Reference to Different Distributions
Authors: Arai Toshihiko , Ando Takao , Kusakabe Atsuko , Ullah Md. Arshad
Published: November 2013

We surveyed plasmids in naturally occurring Vibrio parahemolyticus strains isolated in Japan and Bangladesh. Among the strains isolated in Japan, about half of the strains isolated from stools of patients of domestic diarrhea outbreaks as well as of travelers returning from East Asia were found to have plasmids, but no strains from foods had plasmids. In contrast, among the strains isolated in Bangladesh, none of the four strains isolated from patients had plasmids, but two out of eight strains isolated from water had plasmids, suggesting that plasmids are common in strains from the water in Bangladesh. All plasmids so far reported in V. parahemolyticus were detected in strains isolated from stools of patients. Incidences of plasmids in this organism were not so high in either area. In Japan, all plasmids were detected in strains from human intestines at 37 C, but in Bangladesh, where the temperature is around 30–40 C, the plasmids were detected in strains from the natural environment. These results suggested the possibility that these plasmids can come from different bacteria under rather high temperatures and that incidences of plasmids are influenced by the incidences of plasmids in bacteria present in the vicinity of V. parahemolyticus strains. None of these plasmids were found to have any relation to the biological characters tested.
Adjuvant Activity of Klebsiella O3 Lipopolysaccharide: No Contribution of Proteins to the Expression of Adjuvant Activity
Authors: Kato Nobuo , Kido Nobuo , Ohta Michio , Naito Setsuko , Hasegawa Takaaki , Mori Masashi , Agata Norio , Nakashima Izumi , Kuno Tsuneharu
Published: November 2013

Previously we found that Klebsiella O3 lipopolysaccharide (KO3 LPS) isolated from culture supernatant of strain Kasuya (O3:K1) or its decapsulated mutant strain LEN‐1 (O3: K1–) exhibited very strong adjuvant activity in augmenting antibody response and delayed‐type hypersensitivity to protein antigens in mice. The preparation of KO3 LPS after deproteinization by four cycles of treatment with chloroform–butanol (5: 1) usually contained a small percentage of proteins and a definite amount of another antigen which was destroyed by heating at 100 C for 1 hr. This antigen proved to be derived from type 1 fimbriae which are responsible for mannose‐sensitive hemagglutination of guinea pig erythrocytes. The preparation of KO3 LPS isolated from culture supernatant of the strains which did not produce type 1 fimbriae exhibited strong adjuvant activity similar to that of the preparation from those which produced them. The preparation of KO3 LPS treated with hot phenol water which is known to remove lipid A‐associated proteins exhibited a similar strong adjuvant activity. The preparation of KO3 LPS after extensive deproteinizing, two cycles of pronase treatment followed by ten cycles of treatment with chloroform–butanol, no longer contained detectable amounts of proteins and the fimbrial antigen, but this preparation also exhibited similar strong adjuvant activity. Moreover, there was no difference in strength of the adjuvant activity between the preparation of KO3 LPS isolated from culture supernatant and that isolated by the phenol method from bacterial cells. The present study demonstrates that the strong adjuvant activity of the preparation of KO3 LPS does not depend in any way on proteins contaminating the preparation.
Incompatibility Group K Plasmids in Bacteria Isolated from a Urinary Tract Infection
Authors: Araki Yasuhisa , Inoue Matsuhisa , Mitsuhashi Susumu
Published: November 2013

Citrobacter freundii and Klebsiella pneumoniae were concurrently isolated from a patient with a urinary tract infection. Transferable drug resistant plasmids were isolated from both strains, pMS434 and pMS435. These plasmids belonged to incompatibility group K and both carried genes governing resistance to various aminoglycoside antibiotics, i.e., kanamycin, gentamicin C complex, streptomycin, and 3′, 4′‐dideoxykanamycin B, in addition to those governing resistance to sulfanilamide and ampicillin. They inactivated kanamycin, gentamicin C complex and 3′,4′‐dideoxykanamycin by adenylylation and kanamycin by phosphorylation. Electron microscopic observations disclosed that the molecular weights of the plasmids were about 67.8 megadaltons. These results indicated the similarity in genetic constitution of the two plasmids. This was the second isolation of incompatibility group K plasmids, following that reported by Hedges and Datta.
Fusion of Sendai Viruses or Subviral Envelope Components with Chicken Erythrocytes Observed by Freeze‐Fracture Electron Microscopy
Authors: Hosaka Yasuhiro , Yasuda Yuko , Fukai Konosuke , Ikeuchi Yoko
Published: November 2013

Four different types of envelope of Sendai virus or subviral components, that is, infectious and non‐infectious virions, reassembled envelope particles (REP), and Tween‐ether‐treated envelope fragments (TE), were studied comparatively for membrane interactions with chicken erythrocytes by freeze‐fracture electron microscopy, specifically for membrane alteration by envelope fusion.

The freeze‐fracture replicas of the attachment of the four envelopes in the cold exhibited a common pattern of impressions with attached envelopes, although the fracture plane traversed from erythrocyte to envelope at the periphery of the contact areas of three of the envelopes but not of TE, where the fracture plane mostly cut only through erythrocyte membranes impressed with TE. The freeze‐fracture replicas of the four envelopes reacting with erythrocytes after a short incubation period at 37 C exhibited distinctive features: infectious virions and REP displayed evidence of envelope fusion, but non‐infectious virions and TE showed a particular pattern of envelope association without fusion. Our data demonstrate that the pattern specific for envelope fusion is the formation of a continuous membrane from envelope to cell membrane in a cross fracture of an erythrocyte.
Biological Activities of Guinea Pig Mitogenic Factor from Immune Lymphocytes Stimulated with Antigen
Authors: Iribe Hideaki , Koga Toshitaka
Published: November 2013

Mitogenic factor was produced by sensitized guinea pig lymph node cells stimulated with a specific antigen. Both T lymphocytes and macrophages were required for the production of this factor. The culture supernatant of lymphocytes containing the mitogenic factor exhibited a strong helping effect on the proliferative response of T lymphocytes to phytohemagglutinin (PHA). Mitogenic factor and the factor with the helping activity coeluted in the molecular weight range of 25,000‐35,000 daltons in gel filtration. Furthermore the fraction containing mitogenic factor was found to support the proliferation of lymphoblasts induced by PHA or antigen, suggesting that the mitogenic factor may be the guinea pig equivalent of T cell growth factor (TCGF) reported in the mouse, rat, and human.

On the other hand, the T cell‐activating monokine of the guinea pig, possessing the helping activity for the proliferative response of T lymphocytes to PHA, never exhibited TCGF‐like activity.
Evidence for Generation of Suppressor T Cells in Mycobacterium lepraemurium‐Infected CBA/J Mice, a Mouse Strain Highly Susceptible to the Infection
Authors: Saito Nanako , Hirooka Yoshiko
Published: November 2013

Susceptibility to Mycobacterium lepraemurium (MLM) infection markedly differed between two mouse strains, CBA/J and C57BL/6. CBA/J mice showed high susceptibility to MLM infection and developed either very weak or no delayed‐type hypersensitivity (DTH) to MLM antigen after the injection of MLM. In contrast, C57BL/6 mice, which were resistant to MLM infection, showed significant DTH reaction to MLM antigen after the injection.

Treatment of CBA/J mice with cyclophosphamide (Cy) conferred significant resistance to MLM infection on the CBA/J mice, and the treated mice developed a strong anti‐MLM DTH response after the MLM injection. When spleen cells from MLM‐infected CBA/J mice were transferred to Cy‐treated and MLM‐infected syngeneic mice, the anti‐MLM DTH reaction of the recipient mice was suppressed. Treatment of the spleen cells to be transferred with anti‐Thy‐1.2 antibody or anti‐I‐Jk antiserum plus complement abrogated the suppressive activity. Thus, it is suggested that the high susceptibility of CBA/J mice to MLM infection is due to the generation of Cy‐sensitive, I‐Jk‐positive suppressor T cells after infection with MLM.
Translation of Human Interleukin 2 mRNA in Xenopus laevis Oocytes
Authors: Hirano Toshio , Sudo Tetsuo , Teranishi Tsuyoshi , Fujimoto Kouji , Iizuka Masahiko , Kobayashi Sigeyasu , Onoue Kaoru
Published: November 2013

Poly(A)‐positive mRNA extracted from tonsillar mononuclear cells stimulated with phytohemagglutinin‐M and 12‐o‐tetradecanoyl phorbol 13‐acetate was successfully translated into biologically active interleukin 2 (IL‐2) in Xenopus laevis oocytes, and secreted into the incubation medium. In control experiments, the extract of oocytes injected with either poly(A)‐negative RNA or buffer did not show any IL‐2 activity. By sucrose density gradient centrifugation analysis, IL‐2 mRNA was found as a single peak corresponding to a sedimentation coefficient of 10‐11S.
Delayed Hypersensitivity in Murine Salmonellosis: Specificity of Footpad Reaction in Mice Infected with Rough Mutants of Salmonella typhimurium
Authors: Cho Norio , Saito‐Taki Tatsuo , Nakano Masayasu
Published: November 2013

Delayed type (footpad) hypersensitivity (DTH) in BALB/c mice immunized with rough mutant strains of Salmonella typhimurium LT2 was examined. Injection of live organisms of an Rb mutant TV148 strain induced DTH in mice, while injection of the heat‐killed organisms did not. The mice immunized with live organisms of the Ra, Rb, Rc, Rd, and Re mutant strains showed positive footpad reactions to the heat‐killed cell antigen of LT2 (wild type) strain. The mice immunized with the Rb mutant strain also showed positive footpad swellings in response to heat‐killed cell antigens of S. paratyphi A, S. paratyphi B, S. typhi, S. enteritidis, and S. cholerae‐suis. Furthermore, positive reactions to antigens of Escherichia coli and Shigella flexneri were seen in the TV148‐immunized mice, but the mice did not respond to heat‐killed organisms of Pseudomonas aeruginosa or Staphylococcus aureus. The cross‐reactive footpad reaction to E. coli could be transferred adoptively with T cells prepared from the spleens of TV148‐immunized mice into syngeneic recipients. These results suggest that the cross‐reactive DTH antigen(s) is widely distributed among related organisms such as Shigella and Escherichia.
Increased Susceptibility to Escherichia coli Infection in Mice Pretreated with Corynebacterium parvum
Authors: Yoshikai Yasunobu , Miake Shunji , Sano Masatoshi , Nomoto Kikuo
Published: November 2013

The contribution of activated macrophages to protection against Escherichia coli was studied in mice treated intravenously with Corynebacterium parvum 7 days before infection. C. parvum‐treated mice showed increased phagocytic activity and enhanced resistance to Listeria infection. In contrast, these mice showed increased susceptibility to a subsequent challenge with E. coli that correlated closely with a reduction in the LD50 of lipopolysaccharide (LPS) in these mice. The peritoneal macrophages obtained from C. parvum‐treated mice had a strong ability to phagocytize and kill E. coli in in vitro experiments. A rapid decline in the number of bacteria in the liver of C. parvum‐treated mice was observed in the early period of infection. However, the number of bacteria in liver and spleen increased progressively to a lethal dose from 6 hr after infection. At this time, a significant increase in β‐glucuronidase, a lysosomal acid hydrolase, was found in the serum of these mice. In vitro experiments revealed that the peritoneal macrophages from C. parvum‐treated mice were highly susceptible to the cytotoxic effect of LPS after 6 hr of incubation with LPS. It is suggested that the hypersensitivity of activated macrophages to the cytotoxic effect of endotoxin derived from E. coli may be partly responsible for the increased susceptibility of C. parvum‐treated mice to E. coli infection.
Stimulation of Locomotion of Peripheral Blood Monocytes by Human Plasma Fibronectin
Authors: Yonemasu Kunio , Nakanishi Akira , Sasaki Takako , Kashiba Syuzo
Published: November 2013

The motility of human peripheral blood granulocytes and monocytes in response to human plasma fibronectin was quantified by an in vitro assay using blind‐well chemotaxis chambers. Purified fibronectin under nondenaturing conditions produced increased migration of granulocytes only at concentrations higher than 100 nm, and induced increased chemotactic and random locomotion of monocytes at concentrations higher than 0.1 nm. The monocyte migration‐inducing activity of fibronectin was concentration dependent, and was strongly inhibited by low concentrations of colchicine (100 nm–100 μm). These findings suggest the possibility that plasma fibronectin serves as a chemotactic stimulus for monocytes in vivo and attracts these cells to sites of microscopic tissue injury where plasma fibronectin is deposited.