Atsushi Yoshida

Advantage of gene gun-mediated over intramuscular inoculation of plasmid DNA vaccine in reproducible induction of specific immune responses.

Utilizing a plasmid DNA encoding a single cytotoxic T lymphocyte (CTL) epitope and that encoding ovalbumin (OVA), we compared the reproducibility in the induction of immune responses by gene gun and intramuscular immunization. As compared to intramuscular inoculation, gene gun DNA immunization appeared to bring about highly reproducible and reliable results in the induction of specific CTL and IFN-gamma production to the CTL epitope and production of anti-OVA IgG. The results obtained by intramuscular inoculation vary significantly. Our data shown here strongly suggest that gene gun immunization of skin is a much more reliable method for DNA vaccination to induce effective immune responses in an animal model.

Protective CTL response is induced in the absence of CD4+ T cells and IFN-γ by gene gun DNA vaccination with a minigene encoding a CTL epitope of Listeria monocytogenes

Our work was undertaken to learn the mechanism of induction of protective cytotoxic T lymphocytes (CTL) by gene gun DNA vaccination with p91m encoding an H-2Kd-restricted T cell epitope of listeriolysin O (LLO). Vaccination with p91m induced vigorous antigen-specific CD8+ CTL that produce IFN-gamma and was able to confer partial protection against listerial challenge. However, the p91m-induced protective immunity was revealed to be independent of the IFN-gamma and CD4+ T cell help. The CTL induction is also suggested to require neither adjuvant activity of the plasmid used nor IFN-gamma. The data may be feasible for the design of CTL inducing vaccines in various immunodeficiencies.

Induction of protective immunity to Listeria monocytogenes with dendritic cells retrovirally transduced with a cytotoxic T lymphocyte epitope minigene

ABSTRACT In the present study, we developed a cytotoxic T lymphocyte (CTL) epitope minigene-transduced dendritic cell (DC)-based vaccine against Listeria monocytogenes. Murine bone marrow-derived DCs were retrovirally transduced with a minigene for listeriolysin O (LLO) 91-99, a dominant CTL epitope of L. monocytogenes, and were injected into BALB/c mice intravenously. We found that the DC vaccine was capable of generating peptide-specific CD8+ T cells exhibiting LLO 91-99-specific cytotoxic activity and gamma interferon production, leading to induction of protective immunity to the bacterium. Furthermore, we demonstrated that the retrovirally transduced DC vaccine was more effective than a CTL epitope peptide-pulsed DC vaccine and a minigene DNA vaccine for eliciting antilisterial immunity. These results provide an alternative strategy in which retrovirally transduced DCs are used to design vaccines against intracellular pathogens.

Biased T cell receptor Vbeta gene expression in bronchoalveolar lavage fluid from Japanese patients with sarcoidosis.

Sarcoidosis is believed to be one of the T cell‐mediated granulomatous diseases with unknown aetiology. We attempt to search for the causative T cell clones of sarcoidosis.

Defining the taxonomic status of Streptococcus suis serotype 33: the proposal for Streptococcus ruminantium sp. nov.

To clarify the taxonomic classification of Streptococcus suis serotype 33, we performed biochemical and molecular genetic analyses using isolates (GUT-183, GUT-184, GUT-185, GUT-186, GUT-187T, GUT-188, GUT-189, GUT-190, GUT-191, GUT-192 and GUT-193) from bovine endocarditis. A comparative sequence analysis showed 99.2-100u200a% sequence similarity among the reference strain of S. suis serotype 33 and our isolates for the 16S rRNA gene. These similarities were higher than those between the isolate GUT-187T and S. suis and other streptococci. Comparison of sodA genes also showed high degrees of similarities among the reference strain of S. suis serotype 33 and our isolates (99.7-100u200a%), which were higher than those between the GUT-187T and S. suis and other streptococci. DNA-DNA relatedness among three isolates (GUT-186, GUT-187T, the reference strain of S. suis serotype 33) was over 76.7u200a%. In contrast, the relatedness between GUT-187T and the other streptococcal species (S. suis, Streptococcus parasuis, Streptococcus acidominimus and Streptococcus porci) was 8.4-24.9u200a%. Phylogenetic analyses showed that the isolates did not affiliate closely to any known species of the genus Streptococcus. Moreover, GUT-187T could be distinguished from S. suis and other closely related species of genus Streptococcus using biochemical tests. On the basis of the phenotypic and molecular genetic data, we propose that the isolates of S. suis serotype 33 should be classified into the genus Streptococcus, Streptococcus ruminantium sp. nov. with the type strain GUT-187T (=DSM 104980T=JCM 31869T).

Fatal fungal endocarditis by Aspergillus udagawae: an emerging cause of invasive aspergillosis

Aspergillus udagawae has morphological similarities to Aspergillusfumigatus; however, it shows a low susceptibility to common antifungal drugs and poor in vitro sporulation. We present the first reported case of infectious endocarditis caused by A. udagawae. An awareness of this newly described Aspergillus species is vital for further clarification.

Purulent lymphadenitis caused by Staphylococcus argenteus, representing the first Japanese case of Staphylococcus argenteus (multilocus sequence type 2250) infection in a 12-year-old boy

Staphylococcus argenteus is a novel species separated from a strain of coagulase-positive, non-pigmented S.xa0aureus. Although S.xa0argenteus has been reported to occur globally, multilocus sequence type (ST) 2250 is mainly found in Northeastern Thailand. Because conventional biochemical testing misidentifies this pathogen as S.xa0aureus, multilocus sequence typing (MLST) or nucA sequencing is recommended to distinguish between S.xa0argenteus and S.xa0auereus. The patient was a previously healthy 12-year-old boy who was admitted because of right inguinal lymphadenitis and cellulitis. Although intravenous cefazolin was administered, his lymphadenitis worsened and formed an abscess on day 6 of hospitalization. Incision and drainage were performed on day 7 of hospitalization. Cefazolin was changed to oral cefaclor, and the patient was successfully treated over a period of 5 weeks. No recurrence was observed throughout 12-months of follow-up. He had a history of right axillary lymph node abscess 2 months before this admission, which was successfully treated with incision, drainage, and antibiotic therapy. He has lived in Japan since birth and never traveled abroad. He had no opportunity to interact with foreigners. His immune function, especially neutrophil function, was tested and we did not find any dysfunction. First, methicillin-sensitive S.xa0aureus was misidentified from the abscess culture. Subsequently, the causative agent was re-identified as S.xa0argenteus ST2250 based on MLST. To our knowledge, this is the first case of S.xa0argenteus ST2250 infection in Japan. This pathogen should be taken into consideration in the diagnosis if the patient has atypical non-pigmented S.xa0aureus.

Nuclear moment studies with polarized radioactive nuclear beams

Magnetic dipole and electric quadrupole moments of nuclei in the light-mass neutron-rich region have been studied by taking advantage of spin-polarized radioactive nuclear beams that have been obtained from the projectile fragmentation reaction. Analyses of the results reveal a few interesting phenomena characteristic of nuclear structures in this region. In particular, we report in some detail the latest result on the magnetic moment of the 17C ground state. The distinctly small value of the g-factor obtained, |g(17C)|=0.5054±0.0025, clearly excludes a Iπ=1/2+ candidate for the spin-parity assignment of this marginally bound nucleus, providing a reasonable account of the non-halo nature reported in recent breakup reaction experiments. Finally, future plans at the upcoming radioactive beam facility presently under construction at RIKEN are briefly mentioned.

Field-driven transition in the Ba

Through 2.6 GeV U irradiations, we have induced bimodal splayed columnar defects in Ba

_{1-x}

_{1-x}