bluuebunny/crossref_metadata_2025 huggingface dataset:
Micro‐Neutralization Test for Mumps Virus Using the 96‐Well Tissue Culture Plate and PAP (Peroxidase‐Anti peroxidase) Staining Technique
Authors: Okuno Yoshinobu , Yamanishi Koichi , Lwin Soe , Takahashi Michiaki
Published: November 2013

A rapid micro‐test method for mumps virus neutralization was developed. In this method, a 96‐well tissue culture plate was used for preparation of cell monolayers and the PAP staining technique was used for visualization of mumps virus infected cells. Clusters of infected cells were observed as a focus and the numbers of foci could be counted by the naked eye 2 days after the infection. A linear relationship between virus dilutions and focus numbers was observed. When neutralizing antibodies in sera from cases of natural mumps infection were assayed, a good correlation was observed between those obtained by the focus reduction method applying the micromethod and those obtained by the ordinary plaque method. Our results indicate that this micromethod is useful in mumps virus neutralization tests and it has many advantages over other methods previously reported.
Increased Multiplication of Dengue Virus in Mouse Peritoneal Macrophage Cultures by Treatment with Extracts of Ascaris‐Parascaris Parasites
Authors: Wiharta Agus S. , Hotta Hak , Hotta Susumu , Matsumura Takeo , Sujudi  , Tsuji Moriyasu
Published: November 2013

Methylcellulose‐elicited peritoneal macrophages from BALB/c mice were cultivated in vitro and inoculated with dengue virus (DV). At intervals thereafter portions of the culture fluids were taken and titrated for viral infectivity. Extracts from Ascaris suum and Parascaris equorum, either crude or Sephadex G‐100 fractionated, were examined for effects on the multiplication of DV. The macrophage cultures treated with the above substances produced larger amounts of DV compared with untreated control cultures. The enhancing effect of the substances depended on doses added and duration of treatment and was suppressed by co‐treatment with carrageenan, a specific macrophage‐inhibiting agent, but was not related to the viability of cultured cells. In fluorescent antibody (FA) as well as infectious center assay experiments, it was shown that the DV‐infected cells were found more frequently in treated cultures than in untreated control cultures. In the treated cultures phagocytosis by cultured cells was also of a higher magnitude than that in untreated cultures. In cocultures of macrophages and splenocytes from the same line of mice, no additive effect of splenocytes was noted. The limulus amebocyte lysate clotting enzyme reaction (Limulus test) indicated that involvement of bacterial lipo‐polysaccharides in the enhancement phenomena was negligible. The data so far obtained suggest that the enhancing effect was due to direct action of the parasitic extracts on macrophages. Four Sephadex G‐100 fractions from the crude extracts showed similar activities; however, the effects of fractions I and III appeared to be comparatively strong. Significance of the findings in relation to the pathogenesis of DV infection was discussed.
Dual Effects of OK‐432 on Mitogenic Response of Splenocytes to Concanavalin A
Authors: Tomioka Haruaki , Saito Hajime , Nagashima Kiyofumi
Published: November 2013

OK‐432, a streptococcal preparation, was studied for its effect on the concanavalin A (Con A)‐induced mitogenesis of the host spleen cells. When mice were given a single intraperitoneal injection of OK‐432, there was a substantial increase in the mitogenic response of splenocytes, whereas multiple injections conversely resulted in a marked reduction of the mitogenic response, when the spleen cells were cultured at high cell densities of over than 5 × 105 cells/well. The reduced Con A‐responsiveness in the latter was not restored by mixing spleen cells from mice given multiple OK‐432 injections with those from normal mice. Moreover, splenic macrophages from OK‐432‐injected mice exhibited marked inhibitory activity against Con A‐mitogenesis of normal splenocytes, while normal splenic macrophages failed to show such an effect. Splenic T cells from OK‐432‐injected mice also showed an inhibitory activity against Con A‐mitogenesis of normal splenocytes and similar activity was also noted in normal splenic T cells. Therefore, the OK‐432‐spleen cells contain two types of suppressor cells; one is a newly elicited suppressor macrophage and the other is a suppressor T cell supposedly resident also in normal spleen cells. In the OK‐432‐injected spleen cells, accessory cell function for T cell Con A‐mitogenesis was markedly reduced. On the other hand, it was noted that the interleukin 2‐producing ability of the OK‐432‐splenocytes was augmented more than that of normal splenocytes, indicating that multiple OK‐432 injections also cause an increase in the helper T cell activity of the host spleen cells.
Potentiality of Artificial Sea Water Salts for the Production of Carrageenase by a Marine Cytophaga sp.
Authors: Sarwar Golam , Oda Hiroshi , Sakata Taizo , Kakimoto Daiichi
Published: November 2013

Production of an extracellular enzyme complex (carrageenase) was studied by examining cell‐free fluids from cultures of a marine Cytophaga, 1k‐C783, growing on different media.

Among artificial sea water salts, only NaCl and MgCl2 were utilized by the organism to produce carrageenase. The minimal concentrations of suitable com2 binations of NaCl and MgCl2 were found to be 0.05 M NaCl plus 0.25 M MgCl2, and 0.15 M NaCl plus 0.15 M MgCl2. KCl and CaCl2 did not have any role in carrageenase production in ZoBell 2216 E broth medium.

Carrageenase was synthesized continuously within the resting cells and was released from the cells as well as in the growing cells, when nutrient had been supplied.
Abnormal Glucose Metabolism in Murine Cytomegalovirus (MCMV)‐Infected Mice
Authors: Hayashi Kozaburo , Fujisaki Yoko
Published: November 2013

Five strains of male and female mice were infected with salivary gland‐passaged murine cytomegalovirus (SG‐MCMV) intraperitoneally. Although none of them became hyperglycemic, 13–30% of BALB/c male and female, ICR/Slc male and female and C57BL/6J male mice had abnormal values in glucose tolerance tests (GTT) 14 to 30 day post infection. Serum insulin levels in BALB/c male mice were low and infective virus was detected in the pancreas during the acute phase of infection. Histopathology showed mild edematous changes with inflammatory cell infiltrations in the pancreas. Viral antigen was located in acinar cells and occasionally in beta cells by an immunofluorescence test.
Role of Interleukin 2 on Enhancement of Concanavalin A‐Induced Human Peripheral Blood Lymphocyte Proliferation by Murine B Cell Mitogens
Authors: Nitta Toshimasa , Konno‐Ejiri Hiromi , Okumura Seiichi , Ozawa Atsushi , Nakano Masayasu
Published: November 2013

Murine B cell mitogens such as bacterial lipopolysaccharide (LPS), butanol‐extracted water soluble adjuvant (Bu‐WSA), dextran sulfate (DS), synthetic muramyl dipeptide (MDP), and its analog MDP‐Lys (L18) do not show any mitogenic ability in vitro on human peripheral blood lymphocytes or mixed cell populations of purified T and B cells obtained from the lymphocytes in an ordinary culture system. However, these mitogens are capable of enhancing the mitogenic effect of concanavalin A (Con A) in the cultures.

In the presence of one of these mitogens, the activity of interleukin 2 (IL 2), but not interleukin 1, in the supernatants obtained from cultures containing Con A‐stimulated T cell and B cell populations was higher than that of control cultures. The role of the newly produced IL 2 in the synergistic effect of the mitogens in human lymphocyte cell cultures was discussed.
Enhancement of Dengue Virus Infection in Cultured Mouse Macrophages by Lipophilic Derivatives of Muramyl Peptides
Authors: Hotta Hak , Sanchez Luis Fernando , Takada Haruhiko , Homma Morio , Kotani Shozo
Published: November 2013

Dengue virus multiplication in cultures of a murine myelomonocytic cell line (WEHI‐3) as well as mouse peritoneal macrophages was enhanced by treatment of the cells with lipophilic derivatives of muramyl peptides for 2 or 3 days before virus inoculation, but not for 2 hr before virus inoculation or during the adsorption period. The infection‐enhancing activity of the materials was dependent on their chemical structure, correlating with their immunoadjuvanticity. The infection enhancement in WEHI‐3 cells was due primarily to an increase in the number of virus‐infected cells which was accompanied by an increased cellular capacity to bind latex particles to their cell surfaces.
Some Properties of Vibrio vulnificus Hemolysin
Authors: Shinoda Sumio , Miyoshi Shin‐Ichi , Yamanaka Hiroyasu , Miyoshi‐Nakahara Noriko
Published: November 2013

Some properties of hemolysin produced by Vibrio vulnificus were investigated. The hemolysin was heat labile, and the hemolytic activity was inhibited by adding cholesterol or divalent cations. Cholesterol inhibited the temperature‐independent hemolysin‐binding step, suggesting that cholesterol made up the binding site of the cell membrane, whereas the divalent cations inhibited the temperature‐dependent membrane‐degradation step. However, the V. vulnificus hemolysin was stable to oxygen and sulfhydryl reagents and was not inactivated by antiserum against streptolysin O, suggesting that the V. vulnificus hemolysin differs from oxygen‐labile hemolysins which bind to cholesterol. The V. vulnificus hemolysin seems to be one of the exceptional cholesterol‐binding hemolysins.
Attachment of Mycoplasma pulmonis to Rat and Mouse Synovial Cells Cultured In Vitro
Authors: Araake Minako , Yayoshi Masumi , Yoshioka Morimasa
Published: November 2013

The attachment of Mycoplasma pulmonis m53 organisms to mouse and rat synovial cells was examined by using the organisms and the synovial cells treated in various ways. M. pulmonis treated with trypsin attached to the synovial cells, but the organisms treated with pronase, formaldehyde, glutaraldehyde, or heat did not. These findings suggest that the sites for binding M. pulmonis to the mouse and rat synovial cells are of polypeptide nature. Treatment of M. pulmonis with sialic acid and treatment of the synovial cell sheets with neuraminidase did not affect the attachment. The synovial cell surface for receptors M. pulmonis organisms would be different from those on respiratory cells or erythrocytes for M. pneumoniae or M. gallisepticum. Even nonviable organisms and M. pulmonis membranes attached to the mouse or rat synovial cells. The nature of the receptor of mouse synovial cells would be different from that of rat cells, since rat cells were affected by treatment with formaldehyde or glutaraldehyde, but mouse cells were not.
Sequential Production of Alpha and Beta Interferons and Gamma Interferon in the Circulation of Listeria monocytogenes‐Infected Mice after Stimulation with Bacterial Lipopolysaccharide
Authors: Nakane Akio , Minagawa Tomonori
Published: November 2013

Sequential production of interferon (IFN)‐α/β and IFN‐γ in the circulation of mice which had been previously infected with viable Listeria monocytogenes was induced by injection of lipopolysaccharide (LPS) derived from Salmonella typhimurium. IFN‐α/β production occurred 2 hr after injection of LPS, thereafter IFN‐γ appeared and the maximum titer was demonstrated at 6 hr. At that time, almost all of the IFN was IFN‐γ. IFN‐γ production in response to LPS was observed from the 5th through the 11th day after infection with Listeria, but it was not demonstrated in either mice infected with lower doses of viable Listeria or mice immunized with heat‐killed bacteria. IFN‐α/β production was not drastically affected by treatment with hydrocortisone, cyclophosphamide, carrageenan, antithymocyte serum, or anti‐asialo GM1 antibody, whereas IFN‐γ production was suppressed by administration of all those agents. Noteworthily, IFN‐α/β, but not IFN‐γ, was produced even 6 hr after stimulation with LPS in cyclophosphamide‐ or antithymocyte serum‐treated mice. IFN‐γ induction by LPS was markedly suppressed in mice in which IFN‐α/β produced by Listeria infection itself had been depleted by treatment with anti‐mouse IFN‐α/β antibody, but it was not inhibited in mice when IFN‐α/β induced not by Listeria infection but by LPS had been depleted by treatment with anti‐mouse IFN‐α/β antibody.
Studies on Pathogenesis of Buffalo Pox Virus in Rabbits: Quantitative Assay of Virus in Different Organs
Authors: Chandra Rajesh , Singh Indra Pal , Garg Shri Krishna
Published: November 2013

The buffalo pox virus was found to multiply in the skin, the primary site of inoculation with an eclipse phase of 12 hr. The virus was then detected in the skin after 15 hr followed by its appearance in regional lymph nodes 36 hr postinoculation. Primary viremia was detected 48 hr postinoculation, followed by detection of virus in the lungs, liver, and spleen. The virus multiplied in the lungs on day 4 and in the liver and spleen on day 5 postinoculation and its release led to secondary viremia. In a follow‐up from day 7 to 14 postinoculation, the virus was detected in the kidneys, stomach, intestines, and gonads.
Monoclonal and Oligoclonal Immunoglobulins Localized in Human Dental Periapical Lesion
Authors: Matsumoto Yukihiro
Published: November 2013

The gamma‐globulin fractions of extracts obtained from 70 periapical lesions were studied by agarose gel electrophoresis and immunofixation. In agarose gel electrophoresis, homogeneous bands in the gamma‐globulin region were found in 25 of 70 specimens (36%). Among the 25 specimens showing homogeneous bands, these bands in six specimens were identified as monoclonal or oligoclonal immunoglobulins. Local production of oligoclonal immunoglobulins was suggested since the corresponding homogeneous bands were not detected in the serum of the same patient. These immunoglobulins may be produced as a result of local immunogenic response against infecting bacteria in periapical inflammation.
Anti‐O‐Phosphotyrosine Antibodies in Human Sera
Authors: Ogawa Ryoichi , Ohtsuka Masahiro , Noguchi Kenichi , Iwamura Kenichiro , Watanabe Yasushi
Published: November 2013

Antibodies reactive with O‐phosphotyrosine (PTYR) were detected in 60 out of 621 inpatients, with high frequencies in hematologic and lung malignancies, hepatic diseases, cerebrovascular diseases, and autoimmune diseases. Affinity‐purified antibodies proved capable of recognizing PTYR‐containing proteins in a human carcinoma cell line, A431, both by immunofluorescent staining and by immunoaffinity chromatography, but had no detectable affinity for phosphorylated serine or threonine, or for the nucleotides tested. In these respects, the antibodies observed in human sera were indistinguishable from anti‐PTYR antibodies raised experimentally in rabbits or mice.
Cloning and Expression of the pnd Gene of R16: Determination of Transcriptional Direction and Evolutionary Analysis
Authors: Sakikawa Tomi , Akimoto Shigeru , Ohnishi Yoshinari
Published: November 2013

The gene promoting nucleic‐acid degradation (pnd) of IncB plasmid R16 was cloned into the vector plasmid pACYC177. The pnd gene was found to be located on a 0.55‐kilobase (kb) AluI‐PstI fragment by constructing subclones carrying various portions of the initially cloned fragment. The direction of transcription of the pnd gene was determined by inserting the gene in both orientations into the lacZ′ gene of the plasmid pUR222. In the recombinant plasmid pCM2, transcription of the pnd gene was controlled by the lac promoter region. Addition of cAMP at 42 C resulted in rapid degradation of stable RNA in cells harboring pCM2. In contrast, no RNA degradation was observed in cells harboring pCM14, which has the same insert as pCM2 but in the opposite orientation.

The equivalent gene, pnd of IncIα plasmid R483, has previously been cloned, and a detailed restriction map of the region has been constructed (Akimoto, S., and Ohnishi, Y. 1982. Microbiol. Immunol. 26: 779–793). We constructed a detailed restriction map of the pnd region of R16 and compared it with that of R483. Restriction analyses revealed a similar structure in these two pnd regions. The results suggest that the pnd genes of R16 and R483 have a common evolutional origin.
Virulence of Chick Embryo Fibroblast‐Passaged Flury HEP Rabies Virus and Its Revertants in Mice
Authors: Arai Yohko T.
Published: November 2013

Chick embryo fibroblast‐passaged Flury high egg passage (HEP) rabies virus failed to kill nude mice or cyclophosphamide‐treated mice when inoculated intracerebrally. The virus regained neurovirulence for adult mice after three passages in mouse neuroblastoma C1300 cells (NA cells). However, even after 20 passages in NA cells, the virulence could not be increased to the level shown by the virus passaged several times in suckling mice.

Some physiological and biological properties of the virus showing and not showing mouse virulence after five serial passages and after one single passage in NA cells, respectively, were compared.
A Field Study of Infection with Human T‐Cell Leukemia Virus among Asian Primates
Authors: Ishida Takafumi , Yamamoto Kohtaro , Ishimoto Goichi , Shotake Takayoshi , Takenaka Osamu , Nozawa Ken , Hayami Masanori , Hinuma Yorio
Published: November 2013

Asian nonhuman primates were surveyed seroepidemiologically for natural infection with human T‐cell leukemia virus (ATLV/HTLV) or a closely related agent. Materials from various primates (three genera [Macaca, Presbytis, and Hylobates], 17 species, totalling 1,079 animals) under natural conditions were obtained in the field study. Virus infection was determined by the indirect immunofluorescence test using HTLV‐specific antigens. Animals seropositive for HTLV were found only among macaques originating from various localities, toque monkeys in Sri Lanka (17.5%), crab‐eating macaques in Thailand (1.3%), stumptailed macaques in Thailand (1.5%), rhesus monkeys in Thailand (3.3%), and Celebes macaques in Indonesia (16.9%). Langurs and gibbons were seronegative. Thus the wide distribution of HTLV in nature among various macaques suggests that the introduction of this virus into primates occurred in ancient times.
Persistence of Mumps Virus in Mouse L929 Cells
Authors: Ito Yasuhiko , Tsurudome Masato , Hishiyama Michiko
Published: November 2013

The characteristics of a persistent infection of L929 cells with mumps virus (MuV) is presented. The persistent infection (L‐MuV cells) was regulated by interferon (IFN) produced endogenously and almost all the properties showed that the carrier culture was maintained by horizontal transmission of the virus. Small‐plaque mutants, but not temperature‐sensitive variants, were selected during the persistent infection.

MuV released from L‐MuV cells (MuV‐pi) replicated efficiently in L929 cells, while infection of L929 cells with the original MuV‐o resulted in an abortive infection. The efficient replication of MuV‐pi in L929 cells can be explained by the findings that MuV‐pi induced IFN more slowly and had lower susceptibility to IFN in L929 cells than MuV‐o did. M protein was synthesized to a considerable degree in MuV‐pi‐infected cells, while it could not be detected in MuV‐o‐infected cells. By contrast, MuV‐pi formed small plaques in Vero cell monolayers and the yield of MuV‐pi in Vero cells was lower than that of MuV‐o. M protein induced by MuV‐pi decayed easily in Vero cells. M protein was considered to be a limiting factor for MuV replication in both cell lines.
A Novel Cell‐Surface Antigen Expressed on Most Leukocytes and a Minor Cortisone‐Resistant Population of Thymocytes in Rats: Characterization by Monoclonal Antibodies
Authors: Matsuura Akihiro , Ishii Yoshifumi , Iwaki Hiroyuki , Kikuchi Kokichi
Published: November 2013

A cell‐surface antigen on rat lympho‐hemopoietic cells was determined by using a monoclonal antibody, R2–1B3 (1B3). The 1B3 antibody, when tested for its reactivity with different hemopoietic cells by cytofluorography with a FACS analyzer, labeled more than 80% of lymph node, spleen, and bone marrow cells and 10–20% of thymus cells. Cytofluorographic analysis performed on purified rat T cells, B cells, macrophages, and granulocytes demonstrated that the antigen defined by 1B3 was readily detectable on all of these cell types, with the greatest expression on B cells. A minor population of thymocytes that were labeled by 1B3 appeared to be cortisone‐resistant and were located mainly in the thymic medulla. These 1B3 positive thymic cells seemed to be functionally more mature than 1B3‐negative thymus cells as suggested by the fact that the cytotoxic treatment of thymus cells with 1B3 antibody and complement (C) resulted in significant reduction of their responsiveness to phytomitogens and lymphokines derived from concanavalin A (con A) activated rat spleen cell cultures. Immunochemical data showed that 1B3 antibody recognized the broad ill‐defined band with a molecular weight of 32K to 47K daltons as estimated by SDS‐polyacrylamide gel electrophoresis. These data indicate that the 1B3 defined antigen is distinct from other, previously reported, antigens on rat lymphoid cells including leukocyte‐common (L‐C) and MRC OX‐22 antigens, and that this 1B3 antibody is a useful reagent for analyzing the intrathymic differentiation of T cells in rats.
Protective Effect of Colostrum in Mycoplasma pneumoniae Infection Induced in Infant Mice
Authors: Katsura Takuya , Kanamori Masato , Ogata Sachio , Kitamoto Osamu
Published: November 2013

The immunological responses and mechanism of maternal immunity in Mycoplasma pneumoniae infection of mice were investigated. ICR female mice, 4 weeks old, and infant mice, 2 to 4 days old, were infected with M. pneumoniae. Anti‐M. pneumoniae antibodies in serum and colostrum were determined by enzyme‐linked immunosorbent assay. The specific IgG antibody production persisted for 9 months or longer in both the young and infant mice. These infected mice were protected from rechallenge with M. pneumoniae. In addition, the infected dams conferred passive immunity on their offspring. The infant mice born to uninfected normal dams were protected from the challenge with M. pneumoniae when fed by infected foster dams. Conversely, the infant mice born to infected dams were not protected from the challenge with M. pneumoniae when the infants were fed by uninfected dams. The specific IgG antibody appeared in serum of infant mice inoculated orally with M. pneumoniae‐infected mouse serum and the infants were protected from challenge with M. pneumoniae, while the infants given protein A‐absorbed serum were not protected from the challenge. These results suggest that one of the factors involved in the resistance of infant mice to M. pneumoniae infection is the specific IgG antibody present in the colostrum rather than the result of transplacental transfer.
Numerical Identification of Slowly Growing Mycobacteria
Authors: Tsukamura Michio
Published: November 2013

Numerical identification using the range of a species defined as (M±2s)% was shown to be suitable for the identification of slowly growing mycobacteria. In the formula, M is the mean matching coefficient for individual strains of each species to the hypothetical median organism pattern of their own species and s is the standard deviation.
Immunomodulation in Mice by Experimental Infection with Yersinia enterooolitica
Authors: Ruiz‐Bravo Alfonso , Jimenez‐Valera MaríA , De Cienfuegos Gerardo Alvarez , Ruiz ConcepcióN , Kouwatli Khalil , Ramos‐Cormenzana Alberto
Published: November 2013

Intraperitoneal infection of mice with two strains of Yersinia enterocolitica resulted in an inflammatory response and immunomodulation which appeared to be related to the invasive properties of the bacteria. The primary antibody response to sheep erythrocytes was enhanced by noninvasive cultures of Y. enterocolitica (serotype O:4–33 grown at 22 C and at 37 C, and serotype O:3 grown at 37 C), when given at the same time or two days after the antigen (invasiveness was tested on HeLa cells). In contrast, invasive cultures of serotype O:3 grown at 22 C, injected three days before the antigen suppressed the antibody response; enhancement was caused by these cultures only when given on the day of immunization. Delayed‐type hypersensitivity to sheep erythrocytes was also suppressed by invasive cultures of Y. enterccolitica. These data indicate that the temperature of growth as well as some serotype‐linked factors play a role in immunomodulation by Y. enterocolitica.
Germination of the Decoated Spores of Bacillus megaterium
Authors: Nakatani Yoshihiro , Imagawa Masayoshi , Takubo Yoshihiro , Nishikawa Jun‐Ichi , Nishihara Tsutomu , Kondo Masaomi
Published: November 2013

Decoated spores of Bacillus megaterium ATCC 12872 were prepared by extracting the inner coat components with an alkaline solution containing sodium dodecyl sulfate and dithiothreitol (SDS‐DTT) from outer coat‐deficient mutant spores, which were produced from one of the mutants isolated and named MAE‐05 by us. The decoated mutant spores germinated as well as the intact spores of the mutant and the parent, indicating that the outer and inner spore cats cannot be essential structures for the initiation of germination.

When the SDS‐DTT‐treated MAE‐05 spores were converted to H‐spores by incubation in citrate‐phosphate buffer (pH 3.5) at 30 C for 3 hr, they lost their germinability by glucose and KNO3. Ca‐spores, prepared by treating H‐spores with 10 mM calcium acetate at 37 C for 60 min, regained the germinability. Experiments on the interaction of 45Ca with the cortex and the inner membrane isolated from H‐spores suggested that the calcium present in the inner membrane might be related to germinability.
Large‐Scale Survey of Hepatitis B Virus Infection in Families
Authors: Kashiwagi Seizaburo , Hayashi Jun , Nomura Hideyuki , Ikematsu Hideyuki , Kajiyama Wataru
Published: November 2013

To investigate HBV transmission in families on three islands in Okinawa, Japan, prevalence of HBV markers in two groups of inhabitants was determined. One group consisted of members of families in which there was at least one HBsAg carrier (carrier families); the other group consisted of members of families in which there were no HBsAg carriers (non‐carrier families). A total of 3,261 serum samples were collected from subjects on Iriomote Island, Hateruma Island, and Yonaguni Island. These samples were tested for HBsAg by reversed passive hemagglutination (RPHA) and for antibody to hepatitis B core antigen (anti‐HBc) by radioimmunoassay. Overall prevalences of HBsAg and anti‐HBc were 8.2 and 65.8 per cent respectively. The prevalence of anti‐HBC among members of carrier families (80.8%) was significantly higher than that among members of non‐carrier families (61.6%) (P< 0.001). The prevalence of anti‐HBc among members of carrier families was higher in all age groups, and was particularly so in children. Within carrier families, the prevalence of anti‐HBc was significantly higher in families in which there was at least one HBsAg carrier with HBeAg (94.5%) than in families with no HBeAg‐positive carriers (76.1%). This difference was especially marked in young children. These data suggest that in families with HBsAg carrier(s), the risk of transmitting HBV to members, particularly to young children, is higher than in families without carriers, and that the risk is further increased in families with HBeAg‐positive carrier(s).
Distinct Reactivities of Four Monoclonal Antibodies with Human Interleukin 2 Receptor
Authors: Tanaka Yuetsu , Tozawa Hideki , Hayami Masanori , Sugamura Kazuo , Hinuma Yorio
Published: November 2013

Two new murine monoclonal IgG1 antibodies, H‐31 and H‐A26, were characterized in comparison with two previously obtained monoclonal antibodies against human interleukin 2 (IL‐2) receptor (IL‐2 R), anti‐Tac and HIEI. In immunofluorescence assays with various human hematopoietic cells, H‐31 and H‐A26 antibodies both reacted with only IL‐2 R‐positive cells, and they precipitated IL‐2 R molecules, glycoproteins with molecular weights of 60K and 53K daltons (gp60/gp53), from human T‐cell leukemia virus type I (HTLV‐I)‐carrying MT‐2 cells, as demonstrated by sequential immunoprecipitation after absorption of IL‐2 R with anti‐Tac. Antibody‐binding competition assays showed that H‐31 and anti‐Tac, and H‐A26 and HIEI, respectively, competed reciprocally in binding to the cells, and that anti‐Tac also inhibited the binding of HIEI but not vice versa. H‐31, like anti‐Tac, strongly inhibited the IL‐2‐dependent proliferation of normal activated T‐cells, absorption of IL‐2 and direct binding of IL‐2 to the cells, while H‐A26, like HIEI, inhibited those processes only weakly. The spectra of reactivities of these antibodies with various simian cell lines derived by HTLV‐I infection were different, as revealed by immunofluorescence studies. Human IL‐2 R was shown to express a unique antigenic determinant, detected with HIEI, that was not detectable in IL‐2 R molecules of Old and New World monkeys, and also to express determinants common to simian IL‐2 R molecules. These observations indicate that H‐31 and H‐A26 recognize human IL‐2 R molecules and that the antigenic sites on the IL‐2 R molecule defined by H‐31, H‐A26, anti‐Tac, and HIEI are different.
Fibrinogen Binding Inhibits the Fixation of the Third Component of Human Complement on Surface of Groups A, B, C, and G Streptococci
Authors: Chhatwal Gursharan S. , Dutra Iveraldo S. , Blobel Hans
Published: November 2013

Effects of fibrinogen binding to M protein‐positive and ‐negative streptococci on fixation of the third component of human complement (C3) were determined. In all test cultures of serological groups A, B, C, and G fixation of C3 was observed in normal human serum as revealed by quantitative fluorescent immunoassay. Fibrinogen binding inhibited the fixation of C3 on streptococci. The degree of inhibition was proportional to the extent of fibrinogen binding. Thus, inhibition of C3 fixation was most pronounced in strongly fibrinogen‐positive streptococci of groups A, C, and G and not demonstrable in fibrinogen‐negative cultures of groups C and G. Trypsinization of the streptococci destroyed their capacity to bind fibrinogen and consequently the inhibitory effects on C3 fixation. The carboxymethylated α and β chains of fibrinogen moderately inhibited C3 fixation whereas γ chain had no influence. These studies may indicate that fibrinogen binding structures other than M protein could also be involved in streptococcal pathogenicity.
Stability of the Hexagonal Lattice Structure Formed by an R‐Form Lipopolysaccharide of Klebsiella: Study of Long‐Range Stability
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Ito Hideo , Naito Setsuko , Kuno Tsuneharu
Published: November 2013

The R‐form lipopolysaccharide (LPS) from Klebsiella strain LEN‐111 (O3‐:K1‐) forms a hexagonal lattice structure with a lattice constant of 14 to 15 nm when it is precipitated by addition of two volumes of 10 mM MgCl2‐ethanol. The stability of this hexagonal lattice structure in long‐term incubation at 4 C was investigated. The hexagonal lattice structure was stable for at least 220 days when the LPS was suspended in distilled water, but it had been disintegrated into a rough mesh‐like structure when the LPS was suspended in 50 mM tris(hydroxymethyl)aminomethane (Tris) buffer, pH 8.5, at 4 C for 60 days. Half of the Mg bound to the LPS was released when the LPS was suspended in Tris buffer for 60 days, whereas Mg was not released when it was suspended in distilled water even for 220 days. By contrast, it was stable for at least 220 days in Tris buffer containing 5 mM MgCl2. The LPS suspended in Tris buffer for 60 days, at which time the structure had been disintegrated, could be restored to the original hexagonal lattice structure within 24 hr by addition of 5 mM MgCl2. From these results it is concluded that the hexagonal lattice structure of the LPS retains long‐range stability if Mg bound to the LPS is not released from the LPS.
Effect of Passive Administration of Alloantiserum Containing Antibody against Putative Acceptor(s) for T Cell‐Replacing Factor (TRF) in the Neonatal Stage on Development of B Cell Activity Responsive to TRF
Authors: Hashimoto Yasuhiro , Tsukada Satoshi , Hamaoka Toshiyuki , Sano Yoshimi , Koyama Nobuo , Takatsu Kiyoshi
Published: November 2013

Previously, we showed that the antiserum raised in male (DBA/2Ha × BALB/c)F1(DCF1) mice (T cell‐replacing factor [TRF]‐low response animals) by immunizing them with activated B cells from BALB/c mice (TRF‐high‐responders) contained antibodies against putative TRF‐acceptor site(s). We have now evaluated the hypothesis that neonatal treatment of mice with the above antiserum suppresses the development of B cells responsive to TRF. Male DCF1 mouse anti‐BALB/c B‐cell antiserum or normal DCF1 mouse serum as a control was injected into BALB/c mice within 24 hr after birth. In the antiserum‐treated mice, no augmented primary immunoglobulin M (IgM) antibody responses to sheep red blood cells (SRBC) were observed under the conditions in which markedly augmented IgM anti‐SRBC responses were induced in control BALB/c mice, suggesting that development of B cells reacting with male DCFi mouse anti‐BALB/c B‐cell antiserum is suppressed by the neonatal treatment with the antiserum. Furthermore, the development of B cell activity responsible for helper factors derived from T cells, such as TRF, was markedly suppressed in the neonatally antiserum‐treated mice, whereas activity of B cells capable of interacting directly with helper T cells through antigen‐bridges was not significantly affected by the same treatment. Such suppression of the B cell activity could be induced only when the antiserum was administered within 48 hr after birth. Moreover, neonatal treatment of mice with the antiserum induced suppressed responsiveness of B cells to a T‐independent type 2 antigen, TNP‐Ficoll. Neither serum‐borne suppressive serum components nor suppressor cells were detected by the system employed. These results support the hypothesis that TRF responsive B cells constitute a subpopulation distinct from the other B cells capable of cooperating with helper T cells via cognate interaction.
Protective Effect of Measles Virus Inoculation on Subacute Sclerosing Panencephalitis Virus‐Infected Mice
Authors: Ohuchi Masanobu , Ohuchi Reiko , Mifune Kumato
Published: November 2013

The Biken strain of subacute sclerosing panencephalitis (SSPE) virus caused a fatal neurologic disease in adult mice after intracerebral inoculation. However, the mice were completely protected from the disease when a high dose of measles virus was given intracerebrally after the SSPE virus infection. The measles virus inoculation induced interferon production and immune responses. An experiment with athymic nude mice showed that interferon and anti‐measles antibody were able to prolong the incubation period of the disease but not to protect the SSPE virus‐infected nude mice from death. For complete protection, T lymphocytes appeared to be essential. The present study suggested that the protective effect of measles virus inoculation is basically due to the induction of immune responses and that SSPE virus infection in mice is susceptible to immune reactions.
Protective Effects of Nasal Immunization in Mice with Various Kinds of Inactivated Sendai Virus Vaccines
Authors: Miyamae Takeo
Published: November 2013

The immunoprophylactic effects of nasal vaccination with 13 different kinds of inactivated Sendai virus vaccines were compared by contact exposure to infector mice. Efficacies of the vaccines were evaluated on the basis of the presence of virus‐infected cells by immunofluorescent examination of the entire respiratory tract, including the nasal mucosa.

A single or double inoculations of B‐propiolactone (0.5%)‐vaccine promoted the infection in the respiratory tract, particularly in the nasal mucosa, whereas three inoculations of B‐propiolactone (0.2%)‐vaccine provided considerable protection throughout the respiratory tract with only slight development of serum HI titer. Formalin (0.1%)‐vaccine and UV irradiated‐vaccine strongly protected the nasal mucosa from infection, but did not sufficiently safeguard the lower respiratory tract even with three vaccinations despite adequate development of serum antibody. Nearly complete protection of the entire respiratory tract was induced with six to eight inoculations of a vaccine treated excessively with both UV rays and 1% formalin, without significant development of serum antibody. Out of eight thermal vaccines, five (inactivated at 23 C, 30 C, 37 C and 7 C, and 30 C and 7 C) provided strong protection against infection when inoculated three times. The others inactivated at higher temperatures (37 C, 50 C, or 60 C) were not so protective. High serum HI titers developed, on the whole, with the drop in the temperature required for inactivating the virus. In eight immune mouse groups in which infection was strongly suppressed in the entire respiratory tract, most of the mice harbored less than 50 viral antigen‐positive cells in their nasal mucosa in the postexposure period. The number of the cells was assumed to be a useful criterion for evaluation of vaccine efficacy.
Protective Efficacy against Group B Streptococcal Infection in Neonatal Mice Delivered from Preimmunized Pregnants
Authors: Itoh Takako , Yan Xiao‐Jie , Nakano Hisao , Yoshioka Morimasa
Published: November 2013

Neonatal mice delivered from mothers preimmunized with heated or formalinized whole cell vaccines of type Ia, Ia/c and III/c group B streptococci were infected with each type of bacteria, and then serum antibodies of mothers and neonates who survived the experiments were measured by enzyme‐linked immunosorbent assay. The relationship between the protectivity in neonate mice and the antibody titers to the type specific polysaccharide antigens and the protein c antigen of their sera were examined.

In the Ia‐immunized group which showed high protection against the type Ia infection, anti‐Ia IgG antibody titers were low, and anti‐protein c IgG antibody was not detected. Type Ia/c and III/c vaccines were highly effective against both type Ia/c and III/c infection, but less effective in type Ia infection. The protein c antigen was identified in both type strains by the double diffusion assay, and the IgG antibodies to the protein c were significantly high in sera of both maternal mice immunized with types Ia/c or III/c organisms and their newborn infants. High titers of the protein c IgG antibody retained 3 to 4 weeks after the last injection of vaccines which corresponded to the period of pregnancy and lactation. Small amounts of IgM antibody to all antigens were detected only in maternal sera.

These results suggest that IgG antibodies to the protein c antigen and to the type‐specific polysaccharide antigens are equally important protective factors which are transferable from preimmunized mothers to their newborn infants through placenta and/or lactation.
A Field Study of Infection with Human T‐Cell Leukemia Virus among African Primates
Authors: Ishida Takafumi , Yamamoto Kohtaro , Shotake Takayoshi , Nozawa Ken , Hayami Masanori , Hinuma Yorio
Published: November 2013

African non‐human primates were surveyed seroepidemiologically for natural infection of human T‐cell leukemia virus type I (ATLV/HTLV‐I) or its closely related virus(es). Materials from three genera (Cercopithecus, Papio, and Theropithecus), four species (grivet monkey, Anubis baboon, Hamadryas baboon, and gelada), totalling 983 animals under natural conditions, were obtained in a field study in Ethiopia. Virus infection was determined by the indirect immunofluorescence test using HTLV‐I specific antigens. Animals seropositive for HTLV‐I were found among grivet monkeys and Anubis baboons including the hybrid offspring between Anubis and Hamadryas baboons but not pure‐Hamadryas baboons and geladas. From these results, the HTLV‐I family was proved to be widespread on the African continent and was regarded as a common retrovirus among catarrhines.
Properties of Monospecific Antibodies to the Glycoprotein of Western Equine Encephalitis Virus
Authors: Yamamoto Kiichi
Published: November 2013

Monospecific (MSp‐) antisera against E1 and E2 glycoproteins of western equine encephalitis (WEE) virus were prepared and examined for binding activities to whole virions, hemagglutination‐inhibition (HI), neutralization (NT) and protection. Both anti‐E1 and anti‐E2 MSp‐antibodies (Abs) showed HI activity, but not NT activity. Both NT‐negative MSp‐Abs protected mice against WEE virus challenge. A competition experiment with monoclonal antibodies showed that these MSp‐antisera appear to lack the antibody population for some epitopes involved in viral neutralization.
Inherited Deficiency of the Seventh Component of Complement Associated with Meningococcal Meningitis: Lack of Serum Bactericidal Activity against Neisseria meningitidis in a Girl with C7 Deficiency and HLA Studies of a C7‐Deficient Japanese Family
Authors: Miyake Takeshi , Ohta Kuniaki , Kawamori Juzou , Hirao Takao , Akagaki Youji , Inai Shinya
Published: November 2013

An 8‐year‐old girl with meningococcal meningitis lacked serum complement activity. The seventh component of complement (C7) could not be detected in her serum by either functional or immunochemical analysis. The levels of the other components were within the normal range. Her serum complement activity was restored by the addition of purified C7. Her fresh serum showed a total absence of bactericidal activity against Neisseria meningitidis, group Y, but her serum bactericidal activity was restored by the addition of purified C7. The restoration of her serum bactericidal activity was completely inhibited in the presence of Mg2+ EGTA. These findings suggest that restoration of the bactericidal activity of her serum against N. meningitidis might be mediated by the specific antibody against N. meningitidis and the reconstituted complement system in her serum.

Heterozygous deficiency of C7 was found in 10 of her family members. Genetic studies showed that the mode of inheritance might be an autosomal codominant trait. No genetic linkage between deficiency of C7 and the HLA system was found.
Ecological Studies of Legionella Species
Authors: Ikedo Masanari , Yabuuchi Eiko
Published: November 2013

The occurrence and viable counts of Legionella pneumophila in acid‐treated water samples of 62 cooling towers on the main island of Japan were determined by inoculating them onto plates of Wadowsky‐Yee‐Okuda (WYO) agar medium. WYO plate cultures of 39 (63%) of the samples yielded L. pneumophila with viable counts ranging from 10 to 104 colony‐forming units per 100 ml.

Of the L. pneumophila isolates, 157 were serologically identified as serogroup 1, and the remaining 21 were agglutinated by serogroup 3 (2 strains) and serogroup 6 (19 strains) antisera. In each culture‐positive water sample, the pH and the number of other bacteria were found not to be statistically significantly correlated with the viable counts of L. pneumophila. However, a higher rate of recovery of L. pneumophila was obtained with the water samples with a smaller number of other bacteria. Practical use of commercially available antialgal or antimicrobial agents was found not to be significantly effective for controlling the occurrence and growth of L. pneumophila in cooling tower water.
Experimental Infections of Dogs with Type C Influenza Virus
Authors: Ohwada Kazuo , Kitame Fumio , Homma Morio
Published: November 2013

A study was performed to determine if type C influenza infection could be established in dogs as a model for human cases. Mongrel dogs were infected with the Ann Arbor/1/50 strain of type C influenza virus and were examined for clinical symptoms, virus isolation and antibody response. After the first exposure to the virus, all infected animals developed nasal discharge and some of them also showed swelling of the eyelids, and suffusion of the eyes with tears and eye mucus, within 1 to 4 days. The animals showed an increase in hemagglutination‐inhibiting (HI) serum antibody, and recovery of the agent from the nasal swabs was successful. The symptoms lasted for as long as 10 days in most infected dogs, which was comparable to our human cases reported previously (Katagiri, S., Ohizumi, A., and Homma, M. 1983. J. Infect. Dis. 48: 51–56). After the second and third virus exposures at intervals of 50 days, all animals developed the same symptoms as those described above and the rise in antibody titer was evident. The virus could be recovered from four of the six dogs 2 to 5 days after the second exposure and from one dog as late as 10 days after the third exposure. Increases in antibody titer in the IgM fraction were observed after every infection. In control dogs which were mock‐infected with UV‐inactivated virus, no symptoms were evident and recovery of the virus was not successful although an increase in HI serum antibody titer was seen. These results show that mongrel dogs are sensitive to type C influenza virus and that repeated infections characteristic of human influenza C can be experimentally produced in dogs.
Serological Differences between Human T‐lymphotropic Virus Type‐I (HTLV‐I) and HTLV‐III/LAV
Authors: Harada Shinji , Purtilo David T. , Koyanagi Yoshio , Yamamoto Naoki
Published: November 2013

Sera from each of five preselected groups of patients with acquired immune deficiency syndrome (AIDS), AIDS‐related complex (ARC), hemophilia, adult T‐cell leukemia (ATL), and healthy controls were examined for antibodies to human T‐cell leukemia (T‐lymphotropic) virus type‐I (HTLV‐I) and HTLV‐III by indirect immunofluorescence (IF) and radioimmunoprecipitation (RIP) methods. All sera from five patients with AIDS, ARC, and hemophilia reacted at titers from 1: 512 to 1: 5,120 with fixed H9/HTLV‐III cells by IF but not with fixed MT‐1 cells carrying HTLV‐I. Similarly, sera from patients with AIDS, ARC, and hemophilia precipitated HTLV‐III‐specific polypeptides of 120K, 46K, and 24K. In contrast, sera from five patients with ATL did not react with fixed H9/HTLV‐III cells, but reacted with fixed MT‐1 cells. Moreover, HTLV‐I‐specific polypeptides of 68K, 28K, and 24K were precipitated with sera from ATL‐patients but not with anti‐HTLV‐III‐positive sera. Recently, we infected HTLV‐I‐carrying MT‐4 cells with HTLV‐III and provoked strong cytopathic effects. This system enabled testing for neutralizing antibodies to HTLV‐III. Neutralizing titers to HTLV‐III of five anti‐HTLV‐III‐positive sera ranged from 1:720 to 1:9,000. In contrast, all five seronegative controls showed no or only low reactivity to HTLV‐III envelope (1:80 and 100). However, three out of five anti‐HTLV‐I‐positive sera exhibited weak cross‐reactivities with HTLV‐III. The reactivities were expressed as <1:160, except for one case (1:720). They were considered to be nonspecific since they were negative for HTLV‐III antibodies in the radioimmunoprecipitation studies.
Muramyl Dipeptide Induces Acute Joint Inflammation in the Mouse
Authors: Koga Toshitaka , Kakimoto Kiichi , Hirofuji Takao , Kotani Shozo , Sumiyoshi Akinobu , Saisho Koichiro
Published: November 2013

Acute joint inflammation was produced in BALB/c mice by a single intravenous injection of synthetic muramyl dipeptide (MDP), its stereoisomers and 6‐O‐acyl derivatives of MDP. Four adjuvant‐active, but not five adjuvant‐inactive MDP analogs induced acute swelling and erythema of the ankles and wrists which were detected around 6–10 hr, reached the maximum severity by 18–24 hr and subsided by days 3 to 4 after injection. Introduction of the stearoyl group, but not the α‐branched long chain fatty acid group into the C‐9 hydroxyl group of MDP enhanced and prolonged the joint lesions compared with MDP.