bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Antigen‐Presenting Cells: Characterization of the Cells in the T‐Lymphocyte Proliferative Response
Authors: Koide Yukio , Awashima Fumiyoshi , Akaza Tatsuya , Yoshida Takato O.
Published: November 2013

Human antigen‐presenting cells (APC) which present the antigen to T lymphocytes resulting in a T‐lymphocyte proliferative response were found among peripheral mononuclear cells (MNC), by employing purified protein derivative (PPD) as soluble antigen.

To assess the adherence capacity of human antigen‐presenting cells, MNC were separated by plastic Petri dishes or nylon wool columns. Plastic nonadherent cells were almost equivalent to unseparated cells in antigen‐presenting ability. Plastic adherent cells, however, showed better antigen‐presenting ability than unseparated cells. On the other hand, cells passed over nylon wool columns showed essentially no ability to present PPD to T lymphocytes. Removal of phagocytic cells by carbonyl iron resulted in about 50–70% reduction in antigen‐presenting ability. Carrageenan, which is known to be toxic to macrophages, had no effect on APC.

By using both rabbit anti‐human Ia‐like antiserum and alloantiserum specific for HLA‐DR phenotype and complement, it was shown that APC possessed Ia‐like antigens, whereas they did not bear surface immunoglobulins.

These results indicate that the human APC is probably a cell in the monocyte‐macrophage lineage.

Allogeneic MNC were used as APC in order to determine whether any genetic restriction exists between MNC as APC and responding T lymphocytes. Optimal stimulation was shown to require identity of mixed leukocyte reaction (MLR)‐activating determinants between APC and T lymphocytes. It is, however, obscure whether an HLA‐D region restriction exists in these combinations because PPD‐pulsed allogeneic MNC lost their ability to elicit even MLR. It is possible that this failure to elicit MLR was caused by T lymphocytes among the MNC used as APC.
Lactate Dehydrogenase‐Elevating Agent Is Responsible for Interferon Induction and Enhancement of Natural Killer Cell Activity by Inoculation of Ehrlich Ascites Carcinoma Cells into Mice
Authors: Koi Minoru , Saito Motoo , Ebina Takusaburo , Ishida Nakao
Published: November 2013

Inoculation of Ehrlich ascites carcinoma cells (EAC) into the peritoneal cavities of outbred ddY mice induced interferon (IFN) in the circulation. The maximum titer (1,280 U) was obtained at 24 hr after inoculation. This induced IFN had the characteristics of type I IFN, i.e., stability at pH 2 and lability at 56 C. An increase in natural killer cell (NK) activity was also observed for the first 3 days after inoculation. In addition, plasma lactate dehydrogenase (LDH) activity was elevated in these mice. Inoculation of ascitic fluid or serum of EAC‐bearing mice into normal mice increased plasma LDH activity six‐ to sevenfold over normal levels and elevated activities persisted throughout the life of the mice. These results suggest that the LDH‐elevating agent was responsible for IFN induction and for enhancing NK activity. Because lactate dehydrogenase‐elevating virus (LDV) can be eliminated from tumor cells by passage in vitro, we attempted to grow EAC in tissue culture for several months and re‐examined whether the inoculation of such cells could elevate plasma LDH activity, induce IFN and enhance NK activity. The results showed that inoculation of the passaged cells had no effect on these activities in normal mice. Therefore, we concluded that the IFN inducer was LDV which contaminated the EAC and then enhanced the NK activity. N‐tropic murine leukemia virus also contaminated EAC, but this virus was not responsible because cultured cells of EAC still shed this virus.
Structural Proteins of Poliovirus Type 2 Isolates
Authors: Yoneyama Tetsuo , Hagiwara Akio , Hara Minoru
Published: November 2013

Seventeen strains of type 2 poliovirus isolated from vaccinees and vaccine‐contacts in Japan were analyzed for viral structural proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis. An alteration in electrophoretic mobility of VP1 was observed in most of the strains from paralytic patients and healthy vaccinees. The alteration in VP1 was observed at a high frequency among many strains, but no significant relation between the mobility change of VP1 and antigenic marker or rct/40 marker was observed. No alteration in mobility of other structural proteins was observed among the strains examined except for one with altered VP3. The results suggested that VP1 of Sabin type 2 strain was easily changeable during replication in human intestines.
Electrochemical Modification of Vesicular Stomatitis Virus Glycoprotein by‐Host Cell Transformation
Authors: Miki Takashi
Published: November 2013

Electrochemical properties of the glycoprotein of vesicular stomatitis virus (VSV) grown in Rous sarcoma virus (RSV)‐transformed cells was compared with that of its counterpart grown in nontransformed cells. In DEAE‐Sephadex column chromatography, the glycoproteins of VSV derived from transformed cells appeared more heterogeneous and had a tendency to elute with higher concentrations of NaCl than those from nontransformed cells. In isoelectric focussing, the glycoproteins of VSVs derived from transformed and nontransformed cells appeared as multiple components differing in the isoelectric point, and the glycoproteins from virus from transformed cells had isoelectric points that were more acidic than their counterparts from nontransformed cells. These results show that the glycoprotein of VSV consists of populations of molecules differing in charge and their isoelectric points were shifted to the acidic side by host cell transformation.
Isolation and Characterization of Mycoplasma and Acholeplasma from Apparently Healthy and Diseased (Infectious Sinusitis) Turkeys
Authors: El Sayed Shawki Ali Khider , Chandiramani Nebhan Karamchand , Garg Daya Nand
Published: November 2013

Investigation of 136 turkeys (24 manifesting infra‐orbital sinusitis, 112 apparently healthy) resulted in isolation of 79 strains of Mycoplasma and 4 of Acholeplasma. By the disc growth inhibition test with 16 reference antisera of avian serogroups, 55 strains were identified serologically and 28 remained unidentified. Thirteen strains of Mycoplasma gallisepticum, 1 of M. meleagridis, and 2 of Acholeplasma laidlawii were isolated from turkey sinusitis whereas serogroups C (2), D (19), F (8), M. meleagridis (4), M. anatis (4), A. laidlawii (2), and 28 unidentified strains were isolated from apparently healthy turkeys. Three patterns were recognized on the basis of glucose, maltose, and sucrose, fermentation. The most frequent, pattern I, included 13 M. gallisepticum strains whereas 5 M. meleagridis strains belonged to fermentation pattern III. Isolates were also studied for reduction of tetrazolium, methylene blue, potassium tellurite, resistance to methylene blue and sodium taurocholate, and production of arginine deiminase and “film and spots.” Inoculation of selected isolates into developing chick embryos revealed that 2 A. laidlawii strains were nonpathogenic and 13 M. gallisepticum, 1 serogroup D and 2 serogroup F strains were pathogenic, causing 50–100% mortality. In vitro antibiotic disc sensitivity tests indicated that rovamycin (solubilized spiramycin) may be recommended for turkey mycoplasmosis. Isolation of 2 A. laidlawii strains from turkey sinusitis and 4 M. anatis strains from apparently healthy turkeys appears interesting.
Analysis of Human T‐Cell Antigens by One‐ and Two‐Dimesional Polyacrylamide Gel Electrophoresis
Authors: Fujimoto Junichiro , Ishii Yoshifumi , Kon Shinichiro , Matsuura Akihiro , Kikuchi Kokichi
Published: November 2013

One‐ and two‐dimensional polyacrylamide gel electrophoresis (PAGE) was performed on immunoprecipitates formed between anti‐human T‐cell xenoantiserum (ATS) and cell‐surface glycoproteins of human lymphocytes, that had been radioiodinated by lactoperoxidase and purified on a Lentil lectin‐coupled Sepharose 4B column. In some experiments, the cells were 3H‐labeled by periodate‐tritiated borohydride. ATS that was absorbed with B cells recognized a number of cell‐surface antigens expressed preferentially on human thymus and T cells, with molecular weights of 150K (T150), 94K (T94), 72K (T72), and 65K (T65) daltons. Whereas T150 appeared to consist of multiple components of heavily sialylated glycoproteins and to be expressed largely on thymus and T cells, and to a much lesser extent on B cells, the remaining T94, T72, and T65 glycoproteins seemed to be present on thymus and T cells but absent from B cells. Two‐dimentional PAGE analysis of these T‐cell glycoproteins precipitated by ATS demonstrated that T94 was an acidic glycoprotein with pI of 4, while T72 and T65, the latter being found on thymus and T cells but not on T cell‐type leukemic cells, exhibited marked electric charge heterogeneity with pI ranging from 4 to 7. These data clearly suggest that human thymus and T cells possess a complex antigenic make‐up on their cell surfaces, comparable to that of mouse T cells with a variety of Ly antigen systems.
Drug Resistance and R Plasmids in Bordetella bronchiseptica Isolates from Pigs
Authors: Shimizu Mikio , Kuninori Kenji , Inoue Matsuhisa , Mitsuhashi Susumu
Published: November 2013

A total of 207 strains of Bordetella bronchiseptica isolated from pigs in 1978 and 1979 were tested for drug resistance and for the properties of their R plasmids. Apart from intrinsic resistance to spectinomycin, single (sulfadimethoxine), double (sulfadimethoxine and streptomycin), andt riple (sulfadimethoxine, streptomycin, and ampicillin) resistance were found in 54.1%, 1.0%, and 15.9% of the strains, respectively. All of the triple‐resistance determinants were associated with mercury resistance and were conjugative. pBB1, one of these R plasmids, was identified as Fi− (F) and Spp− (no suppression of phage multiplication) type, and as a member of incompatability group IncP. The single‐ and double‐resistance determinants were nonconjugative. pBB2, one of the double‐resistance determinants, was mobilized by an R plasmid, RP4, with the high efficiency of 80% and at a frequency of 3.3% without cotransfer of RP4. The molecular weight of pBB1 and pBB2 was estimated at 36 × 106 and 13 × 106 daltons, respectively, by electron microscopy and agarose gel electrophoresis. pBB1 had five cleavage sites for EcoRI endonuclease, and four sites for HindIII. pBB2 had two EcoRI sites, one HindIII, and one BamHI site. Cells carrying pBB1 or pBB2 produced enzymic activity that inactivated streptomycin in the presence of ATP.
In Vitro Growth of Mycobacterium lepraemurium under the Influence of Macrophage Cultures
Authors: Matsuo Yoshiyasu , Tatsukawa Hiroshi
Published: November 2013

Elongation and limited multiplication of Mycobacterium lepraemurium was observed extracellularly when the bacilli spotted on a coverslip were placed face to face with cultures of mouse peritoneal macrophages adhering to the inside of a test tube held at an angle of 15°. There was no doubt that certain growth‐promoting but unstable factors were released from the macrophages.
Isolation of an Hswl Nav4 Influenza Virus from a Tufted Duck (Aythya fuligula) in Japan
Authors: Tsubokura Misao , Otsuki Koichi , Yamamoto Hiroshi , Kawaoka Yoshihiro , Nerome Kuniaki
Published: November 2013

A hemagglutinating agent was isolated from a tufted duck captured in Lake Shinji, Shimane Prefecture, Japan, and identified as influenza A by double immunodiffusion tests with the antiserum to influenza A virus ribonucleoprotein. The hemagglutinin of the isolate was antigenically related to that of A/New Jersey/8/76 but was not identical with it. The neuraminidase antigen of this Hswl subtype was closely related to that of A/turkey/Ontario/6118/68 and was shown to be Nav4 subtype. After experimental infection of 5‐week‐old SPF‐chickens with the isolate, virus was recovered from various organs including the brain, despite the absence of signs of disease.
Enhancement of Cytotoxicity of Human Peripheral Blood Lymphocytes by Interferon
Authors: Kato Tsuneo , Minagawa Tomonori
Published: November 2013

Human lung carcinoma cells persistently infected with mumps virus (Pc‐10/MpV) were lysed with human peripheral blood mononuclear leukocytes (PBML) obtained from seropositive donors who had anti‐mumps virus‐neutralizing antibody in their sera. This cellular cytotoxicity was due not to the cytotoxic T lymphocytes but mainly to the non‐T, non‐B cells, possibly related to natural killer (NK) cells. Moreover, it was concerned not with antibody against mumps virus antigens but with alpha‐interferon (IFN‐α) produced in the mixture of human PBML and Pc‐10/MpV cells, since this cellular cytotoxicity was suppressed by anti‐human IFN‐α rabbit serum. Exogeneous IFN‐α augmented the cytotoxicity of non‐T, non‐B cells, not T cells, for the uninfected Pc‐10 cells. IFN‐γ that had been induced by heat‐killed Listeria monocytogenes in PBML had the same capacity to augment NK activity did IFN‐α.
Agglutination, Toxigenicity and Sorbitol Fermentation of Clostridium difficile
Authors: Nakamura Shinichi , Serikawa Toshihiko , Mikawa Masato , Nakashio Satoshi , Yamakawa Kiyotaka , Nishida Shoki
Published: November 2013

A total of 79 Clostridium difficile strains from different sources (50 strains from the fecal specimens of healthy adults, 13 from patients receiving antibiotics without gastrointestinal complications, 13 from antibiotic‐associated pseudomembranous colitis (PMC) or diarrhea patients, and three strains from ATCC) were investigated for agglutinability, using formol‐treated cells as antigen, in relation to toxigenicity. C. difficile strains tested were divided into four serovars, I, II, III, and IV, by the cross‐agglutination test. The agglutinin absorption test revealed that strains of serovar I, agglutinable with high titers (5,120–10,240) to antiserum prepared against a highly toxigenic C. difficile strain, ATCC 17859, possessed the serovar‐specific antigen. All of the strains of serovar I were highly toxigenic and all 13 strains isolated from the fecal specimens of antibiotic‐associated PMC or diarrhea patients belonged to this serovar, whereas 19 (38%) out of 50 strains from healthy adults and four (30.8%) out of 13 strains from patients receiving antibiotics without gastrointestinal complications possessed this antigen. None of the strains of other clostridial species than C. difficile were agglutinated by the three reference antisera used. Further study on the sugar fermentation test disclosed that the sorbitol‐fermenting property of C. difficile is very closely related to the toxigenicity and agglutinability.
Further Studies of the Polysaccharide of Klebsiella pneumoniae Possessing Strong Adjuvanticity: I. Production of the Adjuvant Polysaccharide by Noncapsulated Mutant
Authors: Ohta Michio , Mori Masashi , Hasegawa Takaaki , Nagase Fumihiko , Nakashima Izumi , Naito Setsuko , Kato Nobuo
Published: November 2013

In culture fluid, Klebsiella pneumoniae type 1 Kasuya strain produces polysaccharide exhibiting a strong adjuvant effect. The active substance responsible for the strong adjuvant effect of the polysaccharide is not its acidic polysaccharide fraction (the type‐specific capsular antigen) but the neutral polysaccharide fraction. In the present study, a mutant which did not produce the type‐specific capsular polysaccharide was isolated from ultraviolet‐irradiated cells of K. pneumoniae type 1 Kasuya strain which had been labeled with leucine‐requiring marker by selecting unagglutinable cells with the antiserum to the type‐specific capsular polysaccharide. Serological tests showed that the type‐specific acidic capsular polysaccharide was present neither on the cells surface nor in the culture fluid of the mutant. Electron microscopically, the mutant did not possess any capsular material. On the other hand, nearly an equal amount of neutral polysaccharide antigen was produced in culture fluids of the noncapsulated mutant and the parent strain. The neutral polysaccharide antigen produced by the noncapsulated mutant exhibited the same degree of strong adjuvant effect on antibody response to bovine gammaglobulin in mice as that produced by the parent strain. The relationship between the neutral polysaccharide antigen in culture fluid and the O antigen of K. pneumoniae was discussed.
Comparison of Three Electron Microscopy Techniques for the Detection of Human Rotaviruses
Authors: Trépanier Pierre , Alain Robert , Micusan Valeriu , McLaughlin Bernadette , Berthiaume Laurent
Published: November 2013

Rotavirus detection by direct electron microscopy was compared with direct and indirect immune electron microscopy techniques. The latter two approaches permitted the enumeration of 25 and 103 times more rotaviruses respectively, than direct electron microscopy. Also, 70% and 90% of the virus particles were aggregated by direct and indirect immune electron microscopy techniques respectively, thus facilitating their detection.
Stimulation of Nonspecific Host Resistance to Infection Induced by Muramyldipeptides
Authors: Matsumoto Kensuke , Ogawa Hidemasa , Nagase Osamu , Kusama Tsuneo , Azuma Ichiro
Published: November 2013

The effect of muramyldipeptide (MDP), N‐acetylmuramyl‐l‐alanyl‐d‐isoglutamine [MDP(Ala)], and its analogs on bacterial infection was studied using the experimental model of sepsis infection in mice. Injection of MDP(Ala) gave mice definitive protection against E. coli infection, but only partial protection against P. aeruginosa or K. pneumoniae infection. Several factors influencing the protective activity of MDP (Ala) on E. coli infection were studied, and it was demonstrated that the activity was induced by various routes of administration of MDP(Ala), including the oral route, and was markedly influenced by the bacterial inoculum size. It was also shown that the effective dose of MDP(Ala) was 100 μg per mouse for intraperitoneal, intravenous or subcutaneous injections and 1,000 μg per mouse when administered orally. Furthermore, the optimal interval between MDP‐treatment and infection was 24 hr when the treatment was carried out before infection. Clearance of bacterial cells in blood was observed after E. coli infection in mice treated with MDP(Ala).

The efficacy of MDP(Ala) and two analogs, N‐acetylmuramyl‐l‐valyl‐d‐isoglutamine [MDP(Val)] and N‐acetylmuramyl‐l‐seryl‐d‐isoglutamine [MDP (Ser)], was evaluated for the E. coli infection; MDP(Val) was proven to be slightly less active than MDP(Ala), and MDP(Ser) to be the least effective, although MDP(Val) or MDP(Ser) was reported to have higher adjuvanticity than MDP (Ala) for the development of delayed‐type hypersensitivity.
In Vitro Proliferative Response and Polyclonal Antibody Production in Spleen Cells of Immunologically Defective CBA/N and C3H/HeJ Mice by Water‐Soluble Adjuvant (Bu‐WSA) Extracted from Bacterionema matruchotii
Authors: Okumura Seiichi , Nitta Toshimasa , Nakano Masayasu
Published: November 2013

Mitogenicity and the polyclonal plaque forming cell(PFC)‐inducing property of a water soluble‐adjuvant extracted from Bacterionema matruchotii by butanol (Bu‐WSA) were examined in vitro in the spleen cells of hybrid (CBA/N female × BALB/c male) F1 mice and C3H strain of mice. The hybrid F1 male cells which expressed a CBA/N‐defect were unable to respond to Bu‐WSA, when assessed by the incorporation of [3H] thymidine into the cells and the generation of anti‐trinitrophenyl (TNP)‐PFC or autoantibody PFC defined by the anti‐bromel‐ain‐treated mouse erythrocyte PFC assay. However, hybrid F1 female cells with normal traits responded to Bu‐WSA. Cultured spleen cells of bacterial lipopolysaccharide (LPS)‐nonresponsive C3H/HeJ mice responded to Bu‐WSA as in the case of cells of LPS‐responsive C3H/He mice, and the [3H]thymidine‐uptakes and the numbers of PFC in these culture cells increased. Re‐extraction of Bu‐WSA by phenol did not affect its activities, while the activity of butanol‐extracted LPS on C3H/HeJ cells decreased after re‐extraction by the same procedure with phenol.
Biological and Biochemical Characterization of Macrophage Activating Factor (MAF) in Murine Lymphocytes: Role of Mannopyranosyl Residue of the MAF Molecule in Macrophage Activation
Authors: Fukazawa Yoshimura , Kagaya Keiko , Miura Hiroshi
Published: November 2013

Activation of macrophages with macrophage activating factor (MAF) was evaluated by measuring the intracellular killing activity of murine macrophages against Salmonella typhimurium. Concanavalin A (Con A)‐induced MAF‐rich fraction was obtained by a Sephadex G‐100 column, which contained molecules ranging from 25,000 to 67,000 daltons. The intracellular killing ability of mouse peritoneal macrophages against S. typhimurium was found to be increased by 0.1 m d‐mannose as well as by Con A‐induced MAF‐rich fraction. Both 0.1 m d‐mannose and MAF exhibited a similar timing pattern for macrophage activation. The same concentration of d‐glucose or l‐rhamnose did not change bacterial uptake and intracellular killing by macrophages. Moreover, when MAF‐rich fraction was applied to a Con A‐Sepharose column, a fraction that was adsorbed on Con A and eluted with 0.1 m α‐methyl d‐mannoside exhibited MAF activity. These results suggest the possibility that mannopyranosyl residues in the MAF molecules play an important role as a ligand in macrophage activation.
A Halophilic Vibrio Isolated from a Case of Chronic Cholecystitis
Authors: Konishi Kenichi , Yamagishi Takayoshi , Sakamoto Kenichi
Published: November 2013

We found a halophilic vibrio in B bile from a 75‐year‐old female patient with chronic cholecystitis, and examined its biochemical characteristics. The organisms are gram‐negative short rods or comma shaped, with some ring forms. They have a single polar flagellum, but no capsule. The strains can grow in peptone water with 1.0 to 4.0% NaCl, but not with no NaCl or 6.0% NaCl. The characteristics of the organisms are positive dextrose fermentation, catalase, oxidase, and ornithine decarboxylase, and negative lysine decarboxylase, arginine dihydrolase, and absence of gas from glucose. They are sensitive to 2,4‐diamine‐6,7‐diisopropyl pteridine (0/129). These characteristics indicate that the isolated strain should be a halophilic vibrio. However, no growth on Salmonella‐Shigella (SS), SS with added sucrose and bromcresol purple (SS‐SB), MacConkey's or thiosulfate citrate bile salts (TCBS) agar plates was demonstrated. Nitrate reduction, Simmons' citrate agar, indole, o‐nitrophenol‐β‐d‐galactopyranoside (ONPG), motility and esculin hydrolysis were positive. Urease, gelatinase, Voges‐Proskauer, phenylalanine deaminase and malonate reactions were negative. Acid was produced from amygdalin, arabinose, cellobiose, fructose, galactose, glucose, lactose, maltose, mannitol, salicin, starch, sucrose, trehalose, and xylose, but not from adonitol, dulcitol, inositol, mannose, melibiose, raffinose, rhamnose and sorbitol. From these characteristics the isolate is considered to be not identical with V. parahaemolyticus, V. cholerae, V. vulnificus or other vibrios. It can be presumed that this isolate represents another species of halophilic vibrio.
Suppression of Hen Egg‐White Lysozyme (HEL)‐Specific Delayed‐Type Hypersensitivity Responses in Mice by Suppressor T Cells after Neonatal Administration of Anti‐Idiotypic Antibodies
Authors: Semma Masanori , Amano Tsunehisa , Fujio Hajime , Sakato Nobuo
Published: November 2013

Anti‐idiotypic rabbit antiserum (anti‐Id) directed to the idiotypes of anti‐hen egg‐white lysozyme (HEL) antibody from a single C3H mouse (No. 2) was shown to be capable of recognizing only a fraction of the anti‐HEL antibody populations produced by other C3H mice. Experiments were performed to examine the effect of this particular anti‐Id on the delayed‐type hypersensitivity (DTH) response specific for the same protein antigen. A group of 60‐day‐old C3H mice which had been administered anti‐Id within 24 hr after birth were tested for HEL‐DTH response. The results indicated that the DTH response was completely suppressed by the anti‐Id treatment. The inhibition of DTH reactivity is due to active suppression and involves the generation of suppressor T cells. Thus, the suppression induced with a single injection of anti‐Id was transferable with both spleen cells and thymocytes from mice that received anti‐Id. These suppressor cells are T cells since their ability to suppress DTH is completely abrogated by treatment in vitro with anti‐Thy 1.2 serum and complement.
Intranuclear Incorporation of Thymic Low Molecular Weight RNA by Murine Bone Marrow Immunoblasts and Inhibition of Plasma Cell Formation by a Derivative of Rifampicin
Authors: Nakamura Kunie , Knight Richard A.
Published: November 2013

An in vitro culture system for the proliferation of IgG‐forming plasma cells from mouse bone marrow cultures has previously been described. The present study attempts to elucidate the mode of action of thymic RNA in these cultures. Autoradiography after using radiolabeled thymic RNA showed that radioactive material was mainly incorporated into the nuclei of IgG‐forming plasma cells. No radiolabeled thymic RNA was incorporated into the cells except immunoblasts. The incorporated thymic RNA was acid insoluble and digested by RNase, but resistant to DNase and pronase. Radioactivity in the nucleotide pool after the cells were cultured with radiolabeled thymic RNA was negligible, indicating that reutilization of degraded RNA did not occur in the nuclei of the plasma cells. Moreover, the incorporation of radiolabeled thymic RNA by the cells was not prevented by excess unlabeled nucleosides. Escherichia coli transfer RNA, L‐cell RNA and synthetic polynucleotide poly(A U) were incorporated but were distributed in a different manner in the cells. A derivative of rifampicin, 2′,5′‐dimethyl N(4′)benzyl‐N(4′)[desmethyl]rifampicin (AF/ABDMP), a possible inhibitor of RNA‐dependent DNA polymerases, suppressed both the incorporation of thymic RNA and the differentiation of immunoblasts. AF/ABDMP suppressed DNA synthesis by bone marrow cultures to the same level as those pretreated with anti‐mouse B‐cell antibodies and complement. DNA dependent RNA polymerase activity was observed in the supernatant of bone marrow cultures stimulated by normal syngeneic thymic RNA and human gammaglobulin as antigen. These results imply a possible relationship between B‐cell differentiation and RNA‐dependent DNA polymerases.
Changes in Prostaglandin Levels in Cultures of SV40‐Transformed Macrophage Cell Lines in Relation to Their Phenotypic Expression
Authors: Tanigawa Takahiko , Suzuki Takao , Takayama Hisao , Takagi Atsushi
Published: November 2013

A radioimmunoassay was performed to determine the total amounts of the A, B, and E series prostaglandins (prostaglandins) in culture fluids of simian virus 40 (SV40)‐transformed mouse clonal macrophages, line 28–12, and its subline, 28–12(Ara). When the proportion of phagocytic cells in confluent 28–12 cell cultures increased, the prostaglandin levels in the culture fluids decreased. On the other hand, stably phagocytic 28–12(Ara) cells, which were derived from 28–12 cells and which had a reduced growth rate, did not release prostaglandins under the usual culture conditions; however, when they were treated with lipopolysaccharide or streptococcal preparation OK‐432, large amounts of prostaglandins were released. In contrast, nonphagocytic cell populations in the cultures of 28–12 cells were not responsive to the drug treatment. These results suggest that there is a correlation between the phenotypic change from the nonphagocytic to the phagocytic state and a decrease in prostaglandin levels in culture fluids, and indicate that phagocytic cells are responsive to prostaglandin inducers, whereas nonphagocytic cells are not.
Microbicidal Activity of Bone Marrow Cells. Augmentation of Candida Killing Activity of the Cells by Culturing with Fibroblast Conditioned Medium
Authors: Nozawa Ryushi T. , Yokota Takeshi
Published: November 2013

Mouse bone marrow cells were seeded in 96‐well trays and infected with twofold serially diluted Candida parapsilosis. Outgrowth of the fungi in each well was determined after a 48‐hr incubation period. Freshly collected cells demonstrated a candidacidal activity which increased with increase in the number of cells seeded. When fresh cells were cultivated with conditioned medium of mouse embryo fibroblasts, the candidacidal activity continuously increased for a few days and reached a plateau. The activity was augmented more than 1,000‐fold compared with that of fresh marrow cells. It is suggested that augmentation is due not to the increase in number of effector cells but to the maturation of effector cells. Carrageenan, which is specifically cytotoxic to macrophages, inhibited augmentation of the activity. Therefore, it appears that most effector cells that matured by cultivation with the conditioned medium belong to the monocyte‐macrophage lineage.
Antimicrobial Effect of Human Serum IgA
Authors: Funakoshi Satoshi , Doi Takuji , Nakajima Tsunetaka , Suyama Tadakazu , Tokuda Masao
Published: November 2013

Serum IgA, IgG and colostrum secretory IgA prepared from specimens pooled from a large number of human beings were shown to have measurable levels of antibodies against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, poliovirus, Coxsackie B virus, echovirus and influenza virus. Serum IgA exerted a bacteriostatic effect in vitro on E. coli and P. aeruginosa, which increased in the presence of the iron‐binding proteins lactoferrin and transferrin. This bacteriostasis was reduced when the iron‐binding proteins were saturated with iron. Similar results were obtained with IgG and secretory IgA. The bacteriostatic effect of serum IgA was also shown in vivo, in the peritoneal cavity of mice. The effect was suppressed by iron. Iron‐chelating substances, siderophores, excreted by E. coli diminished the cosoperative bacteriostatic effect of serum IgA and transferrin. Siderophore production by E. coli was inhibited in the presence of serum IgA, but not when serum IgA was deprived of specific antibody by absorption with E. coli. These results indicate that serum IgA has a potent bacteriostatic effect in cooperation with transferrin or lactoferrin because of the inhibitory effect of the specific antibody on siderophore production by E. coli.
Studies on Streptococci
Authors: Kawai Yasuo , Suegara Nobuo , Shimohashi Hirotaka
Published: November 2013

The population levels of bacteria in the contents and the walls of the gastrointestinal tract of gnotobiotic rats inoculated with lactic acid bacteria (streptococci, lactobacilli, and bifidobacteria) from humans and rats were determined. Lactobacilli as well as streptococci isolated from rats colonized in the digestive tracts of the gnotobiotic rats at a high population level, characteristically highest in the stomach. On the other hand, in the rats inoculated with human lactic acid bacteria, streptococci were dominant in the lower tract. The human lactobacilli or bifidobacteria did not colonize when the organisms in each genus were inoculated together with streptococci. However, when all three genera were inoculated together, lactobacilli and bifidobacteria colonized. Observations on the species of streptococci showed that the intestinal type of streptococci was found to colonize at a high population level, but the oral type was not. Strains of the same genus of lactic acid bacteria from humans and from rats showed different colonization patterns.
Subpopulation of T Cells Sensitive to Natural Thymocytotoxic Autoantibody (NTA) of New Zealand Mice: I. Distinct Cytotoxic Sensitivity of Functional T Cell Subsets to NTA and Anti‐Thy‐1 Antibodies
Authors: Nagata Norikazu , Suda Kinya , Maruyama Naoki , Koike Takao , Shirai Toshikazu
Published: November 2013

NZB mice produce a natural thymocytotoxic autoantibody (NTA) capable of specifically injuring thymocytes and T cells. NTA‐reactive antigen (NTA‐A) shows a different density distribution among T cells, and partial killing with NTA and complement can eliminate T cells bearing NTA‐A in high density. Thy‐1 antigen is similar to NTA‐A in this respect. To determine the effects of NTA and anti‐Thy‐1 on distinct functional subsets of T cells, Con A‐induced suppressor T cell (Con A‐Ts) activity against the allogeneic mixed lymphocyte reaction (MLR), responding T cell (TMLR) activity in the allogeneic MLR, and Con A‐induced cytotoxic T cell (Con A‐Tc) activity were examined simultaneously in BALB/c spleen cells before and after partial elimination of NTA‐ and anti‐Thy‐1‐sensitive T cells. Treatment with NTA and complement resulted in a marked reduction in Con A‐Ts activity, a significant increase in TMLR activity and a slight and inconstant decrease in Con A‐Tc activity. Since Con A‐generated Ts were much less sensitive to NTA, the NTA‐sensitive T cells involved in Con A‐Ts activity appear to be precursors or promoters of the Con A‐Ts. In contrast, the precursors of Con A‐Tc seem to be relatively resistant to NTA. The increase in TMLR activity caused by NTA suggests the possibility that NTA is less cytotoxic for TMLR and cytotoxic for some suppressor T cells in allogeneic MLR. The monoclonal anti‐Thy‐1 antibody showed no such preferential cytotoxic effects on the three T cell functions. The NTA‐sensitive T cells, in contrast to anti‐Thy‐1‐sensitive T cells, were reduced gradually during Con A stimulation. All these findings indicate that NTA‐A not only differs from Thy‐1 antigen but that it appears to be a unique T cell antigen.
The Influence of Site of Antigen Deposition on the Local Immune Response in the Mammary Gland of the Ewe
Authors: Watson Dennis L.
Published: November 2013

Experiments were carried out to compare the local antibody responses in mammary glands of ewes immunized by infusion of antigen (killed Brucella abortus) into the lactiferous sinuses (trans‐epithelial presentation of antigen) or injection into the mammary tissue near the supramammary lymph node (interstitial presentation of antigen). Although both methods of antigen presentation resulted in similar antibody levels in blood, infusion of antigen into the lactiferous sinus resulted in significantly higher levels of agglutinating antibody in milk whey than did injection of antigen. When within‐animal comparisons were made, infusion of antigen was also significantly superior to injection of antigen in terms of levels of non‐agglutinating antibody in milk as determined by Coomb's antiglobulin assays. Evidence from immunoglobulin estimations in milk whey suggested that any elevation in concentrations of immunoglobulins (including IgA) in milk from ewes, which had received injections of antigen into mammary tissue, was associated with chronic inflammatory damage to these glands.
Cellulase Enzyme from Aspergillus niger
Authors: Kassim El‐Sayed A.
Published: November 2013

Cellulase enzyme was produced by a selected strain of Aspergillus niger isolated from deteriorated wood and grown on different carbon sources. Filter paper gave the highest yield, followed by carboxymethyl cellulose (CMC). Cellobiose as well as glucose gave a low yield, while the yield from lactose was negligible. The concentration of filter paper cellulose that induced the maximum yield of the enzyme was 1%. Both soluble cellulose (CMC) and cotton cellulose treated with phosphoric acid (swollen) were easily hydrolyzed by cellulase; an increase in cellulase concentration lead to more hydrolysis of CMC and gave linearity in the reaction velocity. At certain concentrations of the enzyme, increase in CMC concentration, (up to 1%) resulted in more reducing sugar. Beyond this point no more hydrolysis occur.
Conjugation of Drug‐Resistance Plasmids from Vibrio anguillarum to Vibrio parahaemolyticus
Authors: Hayashi Fujio , Harada Kenji , Mitsuhashi Susumu , Inoue Matsuhisa
Published: November 2013

Drug‐resistant strains of Vibrio anguillarum, a fish pathogen, were isolated from diseased fish in culture ponds. In investigations of these strains, the transfer of resistance to chloramphenicol, tetracycline, and sulfanilamide from multiple resistant organisms to laboratory recipients was observed. The plasmid from V. anguillarum was stably maintained in both recipient strains of Vibrio parahaemolyticus and Escherichia coli. The plasmids isolated from Vibrio anguillarum belong to incompatibility group C. The molecular weight of these plasmids determined by electron microscopic observation was about 103 to 113 megadaltons.
Binding of HBs Antigen to HeLa Cells by Use of Reconstitution into Liposomes and Fusion by Sendai Virus
Authors: Tanabe Kiyokatsu , Naruto Hiroshi , Shimizu Yoshitaka
Published: November 2013

Hepatitis B surface antigen (HBsAg) was reconstituted into liposomes composed of phosphatidylserin. An isolated membrane fraction of HeLa cells was mixed with the liposomes, then the liposomes were incubated with HeLa cells in the presence of Sendai virus. In this system the attachment of HBsAg to the cell surface was enhanced and became resistant to treatment with trypsin‐EDTA. The presence of the HBsAg on the cell surface was revealed by immunoelectron‐microscopy. This technique may provide a model system for studying immunological reactions to HBsAg‐bearing target cells in vitro.
Mechanism of Antibody‐Mediated Protection against Herpes Simplex Virus Infection in Athymic Nude Mice: Requirement of Fc Portion of Antibody
Authors: Hayashida Isao , Nagafughi Seiho , Hayashi Yoshinobu , Kino Yoichiro , Mori Ryoichi , Oda Hiroshi , Ohtomo Nobuya , Tashiro Akira
Published: November 2013

Experiments were performed to investigate the resistance of the host due to antibody‐mediated mechanisms to herpes simplex virus (HSV) infection. Transfer of hyperimmune anti‐HSV mouse serum inhibited the development of skin lesions and prolonged the survival of lethally HSV‐infected nude mice. Relatively high concentrations of antibody were required to achieve this protection. Antisera prepared in heterologous animals were also effective, while administration of anti‐cowpox virus serum or interferon provided no protection. This type of protection is therefore due to specific antibody and cannot be attributed to interferon. In order to delineate the requirement for antibody in antibody‐mediated protection, human gamma globulin preparations were transferred to lethally HSV‐infected nude mice. Transfer of intact human gamma globulin (GG) was effective in controlling infection. S‐sulfonation of GG did not diminish the protective ability. However, purified F (ab′)2 did not have any protective action even when it was administered frequently to maintain serum neutralizing antibody titer. GG was effective in C5‐deficient mice lethally infected with HSV. These results indicate that in vivo antibody‐mediated protection to HSV infection requires the Fc region of the intact IgG molecule and suggest that antibody‐dependent cell‐mediated cytotoxicity may be operative in vivo.
Synthesis of Glucan on the Cell Surface of Streptococcus mutans: Chemical and Scanning Electron Microscopic Studies
Authors: Tsumori Hideaki , Mukasa Hidehiko , Zinnaka Yutaka
Published: November 2013

The appearance and continuing growth of extracellular material on Streptococcus mutans HS6 cells in sucrose‐containing Merthiolated buffer was observed in a scanning electron microscope and was found to be related to the glucan synthesis on the cell and to adherence of the cell to a smooth surface. Cells grown in broth completely deprived of sucrose by invertase (HS6‐IV) had a characteristic, slightly rugged surface structure. On incubation of HS6‐IV in the sucrose‐containing buffer, a few small globular particles appeared on the surface and grew to an irregular shape (globular to fibrilar) after several hours. The increase in the total glucan content of the cells paralleled the growth of the globular material, to which ferritin‐conjugated anti‐dextran globulin was found to bind. On the cell surface of cells harvested from conventional broth, both small globular and irregular structures, which possibly formed from sucrose in the broth, existed originally and continued to grow during incubation, along with the material newly appearing on the surface. The accumulation of glucan on the cells resulted in their adherence to a glass surface. The inhibition of growth of the extracellular material on the cells by trypsin, dextranase or anti‐glucosyltransferase corresponded to the decrease in glucan synthesis and the loss of adhering ability. These results indicated that the material growing on the cell surface was glucan synthesized by glucosyltransferases.
Effect of Interferon on the Production of HBsAg and Induction of an Antiviral State in Human Hepatoma Cell Line PLC/PRF/5
Authors: Nakajima Yukio , Kuwata Tsuguo , Tomita Yoshimi , Okuda Kunio
Published: November 2013

The effects of human α and β interferons (IFN) on the production of HBsAg by PLC/PRF/5 cells, an HBsAg‐producing human hepatoma cell line, were studied in the exponential and stationary phases of cell growth. When exponential phase cells were treated with 100 or 1,000 U of IFN per ml for 48 hr, the amount of HBsAg in the culture medium decreased. The number of cells and the synthesis of DNA and proteins were also reduced by the IFN treatment. These results suggested that IFN did not affect the production of HBsAg specifically in exponential phase cells. When cells in the stationary phase were similarly treated with IFN, HBsAg production was not inhibited nor did the number of cells decrease.

To examine the antiviral state induced by IFN in PLC/PRF/5, induction of 2′5′‐oligo(A) synthetase and susceptibility to two kinds of viruses were examined. The 2′5′‐oligo(A) synthetase activity was increased in an IFN‐dose dependent manner. Susceptibility to vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV) was decreased by treatment with 10 and 100 U of IFN per ml for 20 hr. It was concluded that IFN‐α and IFN‐β induce 2′5′‐oligo(A) synthetase and the antiviral state, but do not inhibit HBsAg production by PLC/PRF/5 cells.
Effect of Anti‐Ia Serum In Vitro on Lipopolysaccharide‐Induced Proliferative Responses of Murine Bone Marrow and Spleen Cells
Authors: Tanabe Masao J. , Nakano Masayasu
Published: November 2013

The in vitro proliferative response of murine bone marrow cells and spleen cells to bacterial lipopolysaccharide (LPS) and the effect of anti‐Ia serum on the response were studied. The incorporation of [3H]thymidine into cells prepared from bone marrow increased in the presence of LPS, but the addition of anti‐Ia serum to the cultures reduced the incorporation. Pretreatment of bone marrow cells with anti‐Ia serum and complement did not abolish the ability of the cells to respond to LPS, while the same pretreatment destroyed this ability in spleen cells. These results suggest that cultures of Ia‐negative bone marrow cells generate Ia‐positive cells during the culture period, and the Ia‐positive cells are responsive cells to LPS.

The proliferative response of 1‐ or 2‐week‐old spleen cells was easily suppressed by anti‐Ia serum when compared with that of 4‐week‐old spleen cells. Furthermore, the responses of spleen cells obtained from γ‐irradiated and syngeneic bone marrow cell‐reconstituted mice were prominently suppressed by anti‐Ia serum in comparison with that of normal adult spleen cells. These findings suggest that LPS‐responsive lymphocytes in the developmental stage are quite sensitive to anti‐Ia serum. The effect of anti‐Ia serum on the maturation of bone marrow‐derived lymphocytes was discussed.
Effect of Lead Acetate on Superoxide Anion Generation and Its Scavengers in Mice Given Endotoxin
Authors: Sakaguchi Osamu , Abe Hiroharu , Sakaguchi Shuhei , Hsu Cheng‐Chin
Published: November 2013

The administration of endotoxin to mice rendered hypersensitive by lead acetate resulted in profound lipid peroxide formation in the liver 6 hr postintoxication. Endotoxin plus lead acetate administration depressed glutathione peroxidase and superoxide dismutase activities in mouse liver, whereas superoxide anion generation significantly increased in the livers of endotoxin plus lead acetate‐treated mice compared with that in mice treated with endotoxin alone. Serum acid phosphatase and lactate dehydrogenase isozyme exhibited much more leakage in endotoxin plus lead acetate‐injected mice than in sera of mice given endotoxin alone. Nonprotein SH level in the liver was reduced markedly in endotoxin‐lead treated mice compared with those receiving endotoxin alone. The plasma vitamin E level was found to decline by 6 hr postintoxication in both endotoxin‐lead and endotoxin alone‐treated mice, and the transient elevation of the plasma level at 18 hr may be considered to indicate mobilization from other tissues into the blood.
Neutralization of Sendai Virus by the IgG Subclass Antibodies of the Guinea Pig
Authors: Togashi Hiroyuki , Tozawa Hideki
Published: November 2013

A comparative study was made of the neutralizing activities of IgG subclasses IgG1 and IgG2, fractionated from guinea pig antisera against Sendai virus. The yields of IgG2 from the antisera were about 16 times as much as those of IgG1. The neutralizing activity of IgG2 per unit weight was four times as high as that of IgG1. This neutralizing activity of both IgG subclasses was enhanced about 10 times by addition of antibodies to the L‐chain of guinea pig immunoglobulin. It is suggested that, in the complement‐dependent neutralization of the virus, IgG1 and IgG2 activate the complement through the alternative and the classical pathway, respectively.
Further Studies of the Polysaccharide of Klebsiella pneumoniae Possessing Strong Adjuvanticity: III. Augmentation of the Antibody Response to Subcutaneously Injected Sheep Red Blood Cells by the Adjuvant Polysaccharide
Authors: Yokochi Takashi , Nakashima Izumi , Nagase Fumihiko , Kato Nobuo , Ohta Michio , Fujii Yasuaki , Mizoguchi Kenji , Isobe Ken‐Ichi , Saito Mitsuru
Published: November 2013

The adjuvant action of the O3 antigen of Klebsiella (KO3) on the antibody response to sheep red blood cells (SRBC) was elucidated by injecting both KO3 and SRBC subcutaneously at the right inguinal region of SMA mice. We demonstrated that KO3 exhibits a novel ability to augment anti‐SRBC plaque‐forming cell responses in both the local lymph node and the spleen at a relatively late stage of immunization. Escherichia coli lipopolysaccharide, dextran sulfate and concanavalin A showed such an action only minimally. In parallel with the development of the adjuvant action, KO3 definitely activated B cells in the local lymph node polyclonally for either IgM or IgG synthesis, suggesting that the mechanism of the adjuvant action includes direct stimulation of B cells by KO3 at the late stage. Neither increase in trapping of lymphocytes in the local lymph node nor change in tissue distribution of antigen was shown to be primarily involved in the mechanism of the adjuvant action.
Variable Sugar Fermentation by Clostridia
Authors: Nakashio Satoshi , Nakamura Shinichi , Nishida Shoki
Published: November 2013

Forty‐two sugar fermentation characteristics recorded as “d” in the 8th edition of Bergey's Manual of Determinative Bacteriology were reinvestigated by using eight different peptone media and 205 strains of nine species of pathogenic Clostridia. In the absence of sugar, the final pH of 7‐day cultures in the basal medium varied widely depending on the peptone employed, the species tested, and even the strain of the species. In the presence of sugar, the final pH of 7‐day cultures was markedly influenced by these factors. Since the sugars with reactions recorded as “d” were weakly fermented and, consequently, the fermentation reactions were easily affected by cultural variations resulting in strain instability, we employed the pH difference method (ΔpH method) in which the pH difference of 0.5 between cultures with and without sugar was used as the critical level. Thirty‐six (86%) of the 42 sugar reactions recorded as “d” were sorted into “+” or “‐” by the ΔpH method. Six sugars, however, still remained as “d” because of their extremely weak fermentation characters. The use of the ΔpH method in any peptone medium not only minimizes incorrect evaluation but also can provide some distinct criteria for identification of Clostridia.