A protein associated to prodigiosin formation was found in Serratia marcescens. The protein was not found in nonpigmented strains and was correlated with the pigment level. The protein was about 100 kilodaltons (kDa) and was also found in nonpigmented bacteria of the pigmented strain grown in glucose medium, at high temperature, or under anaerobic condition. The 100kDa protein was found not in the outer membrane and the periplasm, but in the inner membrane and/or the cytoplasm. The protein was also found singly or dominantly in pigmentprotein complexes and pigment-localizing vesicles described in previous reports. These results suggest that the 100kDa protein is associated with prodigiosin formation.

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A subtype (CS2) of the colonization factor antigen II (CFA/II) of human enterotoxigenic Escherichia coli (ETEC) was studied. Analysis revealed that CS2-possessing ETEC was predominant among isolates from traveller's diarrhea at Osaka, Japan. TH61 pili produced by a clinical strain (TH61) were purified as a native form to homogeneity by zone electrophoresis and successive column chromatographics on Sepharose 4B and Phenyl-Sepharose CL-4B. It was demonstrated by immunogold staining technique and bacterial agglutination test that antisera against the purified pili of strain TH61 recognized pili of both strain TH61 and strain #C91f, a control strain possessing only CS2 pili. This suggests that TH61 pili purified in this study are CS2 pili. Subunit (pilin) of the purified pili has a molecular weight of about 16, 000. Strains bearing CS2 could attach to human jejunal epithelial cells, and this attachment was inhibited by pretreating the enterocytes with purified pili. These indicate that CS2 pili are a factor responsible for attachment of ETEC bearing CS2 to human intestinal cells.

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Active immunization of mice with high doses of heat-killed unencapsulated strains of Staphylococcus epidermidis, which were grown in brain heart infusion media, protected mice against challenge with encapsulated strains of S. epidermidis. The unencapsulated strains were capable of absorbing the protective antibody in rabbit hyperimmune sera prepared with the encapsulated strains. Also, mice treated with rabbit hyperimmune sera prepared with the unencapsulated strains were protected against challenge with the encapsulated strains. The protective activities of these rabbit hyperimmune sera were assumed to be essentially identical to those of the protective antibody induced by the encapsulated strains.

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The cell wall of Clostridium difficile GAI 4131 was revealed by electron microscopy to have an outer layer composed of a nearly square array and contained the two major proteins with molecular weights of 38kDa and 42kDa. The properties and reassembly of the two major proteins into the regular array were investigated. When the isolated cell walls were treated with hydrophobic bond-disrupting agents or a chelating agent specific for Ca²⁺, the two major proteins were effectively removed and the regularly arranged outer layer disappeared. The amino acid composition of the two major proteins differed from each other. The two major proteins also gave different peptide maps from each other upon proteolysis with Staphylococcus aureus V8 protease. The major proteins solubilized from the isolated cell walls with 8M urea or 4M guanidine hydrochloride could be reassembled into open-ended cylinders possessing the native regular pattern by dialysis against neutral buffer containing 5mM CaCl₂. The reassembled cylinders purified by centrifugation on a Percoll density gradient were composed of almost equal amounts of the 38kDa and 42kDa proteins and freed from the other proteins. These results suggest that the regular array in the outer cell wall layer is constructed from the two major cell wall proteins and requires Ca²⁺ for its assembly.

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In order to determine the transcripts of gpI and gpIV of varicella-zoster virus (VZV), RNA was isolated from human embryonic fibroblast cells infected with VZV and Northern blot analysis was carried out using cloned DNA probes of unique short region including gpI and gpIV genes. The analysis of RNA revealed two discrete transcripts of 3.6 and 2.15 kilobases (kb) and three transcripts of 3.6, 2.9, and 1.6kb which hybridized to DNA probes covering the gpI and gpIV region, respectively. Next, mRNAs were hybrid-selected, translated in vitro and the polypeptide products were immunoprecipitated with antibodies against these glycoproteins. The polypeptides with a molecular weight of 70, 000 (70K) and 37K which were in vitro translational products of mRNA hybrid-selected with the DNA clone covering gpI and gpIV were detected using antibodies against gpI and gpIV, respectively. The result showed that the 70K polypeptide is presumably the translational product of 2.15kb mRNA and the 37K polypeptide is that of 1.6kb mRNA. DNA fragment encoding gpI or gpIV was inserted into vaccinia virus DNA and the recombinant viruses, mO74 (gpI) and mO39 (gpIV), were used for immunological analysis. In consequence, the gpI derived from cells infected with mO74 showed antigenic characteristics similar to those of gpI from VZV-infected cells as determined from the immunoprecipitation pattern, although the molecular weight of each polypeptide was different, and antibody produced in rabbits infected with recombinant virus had a high neutralizing activity, when the reaction was performed with complement. This suggested that gpI plays an important role for protection and recovery from VZV infection.

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Early development of T lineage cells were compared between AKR and C3H mice by using two experimental strategies-neonatal thymectomy (NTx) and bone marrow transplantation (BMT)-between these two strains of mice. After NTx, AKR mice developed less wasting disease and showed better maintenance of several T cell functions. In addition, the response of neonatal spleen cells to PHA and ConA was much greater in AKR mice than in C3H mice. Further, when AKR mice were used as recipients of BMT, cell numbers recovered from thymuses between 2 and 7 weeks after reconstitution were consistently much greater (about 10 times greater) than those from chimeras where C3H mice were used as recipients, regardless of the donor strains of bone marrow cells. However, 4 weeks after BMT the proliferative responses to ConA were consistently higher in the donor-derived thymocytes from chimeras where AKR mice were used as bone marrow donors than in those from chimeras in which C3H were donors. The present findings suggest that these differences may be attributed to characteristics of recipient microenvironment (e.g., thymic stroma) which maintain developing thymocytes and supply them to the peripheral lymphoid tissue. Alternatively the differences may to some degree also be attributable to characteristics of the thymic progenitors themselves, which may determine the rates of maturation of thymocyte functions.

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B220, a pan-B marker, is known to be also expressed on immature T cells of MRL/1pr or other congenic 1pr mice and minor population of immature thymocytes but not on peripheral T cells. In this study, we investigated in vivo the possibility whether B220 is one kind of premature T or prethymic T precursor cell marker as well as a pan-B cell marker. A monoclonal antibody against B220 glycoprotein was intravenously injected every 2 days into various strains of mice. After the third administration of this antibody, thymocytes decreased remarkably compared with those from the rat IgG-treated group, and spleen cells were also reduced significantly. Further, the number of cells expressing Thy-1, Ly-1, Lyt-2, and asialo GM1 (asGM1) in the spleen were significantly reduced. On the contrary, the number of cells expressing surface IgM (sIgM) or B220 were increased by this treatment, especially after the 8th treatment. Some T-dependent immunological functions including mitogenic responses to lectins and cytotoxic T cell activity were markedly suppressed but mixed lymphocyte reaction (MLR) and natural killer (NK) activity were rather augmented. Thus, B220 may be expressed on some kinds of T cell progenitor. Taken together, in vivo treatment with anti-B220 antibody may influence differentiating stages of some T cells from bone marrow progenitors before or just after their homing into the thymus.

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Pertussigen [pertussis toxin (Ptx)] from Bordetella pertussis, when detoxified, induces protection in mice to intracerebral challenge (ic) with virulent B. pertussis. In its native form, minute nonprotective doses promote the development of immunity induced by other antigens of B. pertussis. As little as 4ng of Ptx, given with a nonprotective dose of 8×10⁷ killed cells of the phase III Sakairi strain, promoted detectable protection to is challenge. Native Ptx in doses of 0.4 to 400ng did not protect mice, and vaccines made from strains not producing Ptx induced only weak protection. The marked enhancing action of Ptx was also observed with 5μg of purified filamentous hemagglutinin and with vaccines made from other species of the Bordetella genus, such as B. parapertussis and B. bronchiseptica, but it was not observed with B. pertussis endotoxin. In addition, Ptx was still effective when given as late as 7 days after the vaccine. Antibodies to surface antigens of the challenge strain were demonstrated in sera of mice immunized with vaccines prepared with the different Bordetella species tested, but antibodies to Ptx were detected only in the sera of mice immunized with the wild-type B. pertussis strains. Glutaraldehyde detoxified Ptx does not have this action. Pretreatment of normal mice with Ptx, also enhanced the protective action of a mouse antiserum to a wild-type strain of B. pertussis. These observations show that antigens other than Ptx are responsible for the protection, and that Ptx acts non-specifically to enhance the mouse protective action of those antigens.

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A water-soluble oligosaccharide, N-acetyl chitohexaose (NACOS-6) was able to enhance the protecting effect of BALB/c male mice against Listeria monocytogenes infection, when administered intraperitoneally 24hr before the challenge with this microbe. Significant decrease in number of microbes within the peritoneal cavity, spleen, and liver from the mice of NACOS-6-administered group was not observed 1 day after the infection but 4 days after the infection. Administration of NACOS-6 enhanced the delayed-type hypersensitivity response against sheep red blood cells (SRBC) or heat-killed L. monocytogenes. Splenic T lymphocytes from mice administered NACOS-6 released macrophage activating factor (MAF). These results suggested that NACOS-6 was also able to elevate the function of cellular immunity. Macrophages treated with a combination of NACOS-6 and the culture supernatant of splenic T lymphocytes from mice administered NACOS-6, “NACOS-6 sup, ” were found to exert a fairly strong growth-inhibitory effect on L. monocytogenes. Interferon-γ (IFN-γ) and interleukin 2 (IL-2) were able to enhance the growth-inhibitory effect on L. monocytogenes by the NACOS-6-treated macrophages.

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A 33-year-old man complained of purpura (petechial hemorrhage) in chelidons, poples, axillae, and bilateral chest in addition to other symptoms such as lumbago, arthralgia, muscular pain, and fever. On the next day of the onset, human parvovirus (HPV/B19) antigen and HPV/B19 DNA were detected in his serum, and twelve days later IgM antibody to HPV/B19 became detectable. This case supports the relationship between purpura and HPV/B19 infection.

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