Yasuhiro Shibasaki

Yasuhiro Shibasaki
Nihon University | Nichidai · College of Bioresource Sciences

Ph. D. DVM

About

47
Publications
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939
Citations

Publications

Publications (47)
Article
Full-text available
Unlike mammals, fish express two type II interferons, IFNγ and fish‐specific IFNγ (IFNγ‐related or IFNγrel). We previously reported the presence of two IFNγrel genes, IFNγrel 1 and IFNγrel 2, which exhibit potent antiviral activity in the Ginbuna crucian carp, Carassius auratus langsdorfii . We also found that IFNγrel 1 increased allograft rejectio...
Article
Germinal centers (GCs) or analogous secondary lymphoid microstructures (SLMs) are thought to have evolved in endothermic species. However, living representatives of their ectothermic ancestors can mount potent secondary antibody (Ab) responses upon infection or immunization, despite the apparent lack of SLMs in these cold-blooded vertebrates. How a...
Preprint
Full-text available
The co-evolution between secretory immunoglobulins (sIgs) and microbiota began with the emergence of IgM over half a billion years ago. Yet, IgM function in vertebrates is mostly associated with systemic immunity against pathogens. sIgA and sIgT are the only sIgs known to be required in the control of microbiota homeostasis in warm- and cold-bloode...
Article
Full-text available
Hemoglobin beta (Hbß) is a heme-binding protein capable of oxygen delivery. The oligopeptides derived from Hbβ in fish mucus are active against a variety of gram-negative bacteria and protozoa. To gain information on the physiological and immunological roles of Hbβ in the mucosal tissues of fish, we analyzed changes in Hbß gene expression levels in...
Article
To search immune defense proteins in skin mucus of Japanese flounder fed with a diet containing high concentration of ascorbic acid, we carried out 2D-PAGE and compared the resolved pattern of proteins between control group that fed commercial diet and ascorbic acid supplemented group (AsA group) fed a diet supplemented with high concentration of a...
Article
Full-text available
Background Two species of deer ked ( Lipoptena cervi and L. mazamae ) have been identified as vectors of Bartonella bacteria in cervids in Europe and the USA. In an earlier study we showed that Japanese sika deer ( Cervus nippon ) harbor three Bartonella species, namely B. capreoli (lineage A) and two novel Bartonella species (lineages B and C); ho...
Article
Although mammalian secretory immunoglobulin A (sIgA) targets mucosal pathogens for elimination, its interaction with the microbiota also enables commensal colonization and homeostasis. This paradoxical requirement in the control of pathogens versus microbiota raised the question of whether mucosal (secretory) Igs (sIgs) evolved primarily to protect...
Article
Our previous work has shown the existence of distinct subsets of B cells in vertebrates that are highly phagocytic. To understand further the involvement of B cells in other potential innate immune functions, we performed the first comparative transcriptome analysis on FACS-sorted IgT+ and IgM+ B cells in rainbow trout, a primitive teleost fish spe...
Article
Full-text available
TCR/CD3 complex is composed of the disulfide-linked TCR-αβ heterodimer that recognizes the antigen as a peptide presented by the MHC, and non-covalently paired CD3γε- and δε-chains together with disulfide-linked ζ-chain homodimers. The CD3 chains play key roles in T cell development and T cell activation. In the present study, we found nor or extre...
Data
Immuno-precipitation and Western blot analysis. After immuno-precipitation of the thymus and spleen protein samples with anti-gCD3ε Ab, the proteins were detected by western blotting with anti-gCD3ε Ab. Both samples show some bands around 20–25 kDa expected to be gCD3ε and single band around 55 kDa expected to be heavy-chain.
Data
Protein sequencing by LC-MS/MS. Protein sequencing was determined using a protein band reactive with anti-gCD3ε Ab detected by Western blot. LC-MS/MS revealed 28 amino acid residues (gray highlight) and 16.1% of residues matched with gCD3ε amino acid sequence (A). Mass spectrum and fragmentation tables of each amino acid fragments are shown in (B,C...
Data
Expression analysis of CD3ε in ginbuna tissues by RT-PCR. Total RNA was prepared from peripheral blood leukocytes (PBL), thymus, head-kidney, trunk-kidney, spleen, liver, ovary, intestine, skin, and gill tissues, and used for RT-PCR analysis. ef-1a was used as an internal control. Numbers to the right indicate PCR cycles.
Data
Modulation of CD3ε expression. After allo-antigen stimulation (A) or Edwardsiella tarda infection (B), kidney lymphocyte was stained with anti-gCD3ε as described above and analyzed by FACS. Mean ± SD of more than three independent experiments are shown. Statistical significance was calculated using t tests to each gene (ns, not significant; p > 0.0...
Data
Effect of in vitro culture on the leukocytes composition. Before in vitro (0 h) culture, leukocytes from kidney are composed of 46.2% of granulocytes, 11.1% of monocytes, and 39.3 % of lymphocytes. Similarly, after in vitro (24 h) culture, leukocytes from kidney are composed of with 50.0% of granulocytes, 7.6% of monocytes, and 32.2% of lymphocytes...
Data
Gene expression analysis of T and B cell related genes in sorted CD3ε+ lymphocytes. Spleen cells were stained with anti-CD3ε Ab as described. Lymphocytes fraction from spleen were gated on FS and SS dot plot and anti-CD3ε Ab positive cells were sorted by FACS. Total RNA was prepared from 1×106 sorted cells and used for RT-PCR analysis. mRNA express...
Data
FS and SS dot plots of kidney and spleen leukocytes. Lymphocytes, myeloid cells, and granulocytes were gated on FSClow SSClow, FSChigh SSClow, and FSCmed SSChigh population, respectively (A). Lymphocytes from kidney and spleen were gated on FSClow SSClow population. The percentages of anti-CD3ε pAb positive cells were shown in the histogram (B).
Data
Ginbuna CD3ε sequence. (A) Ginbuna CD3ε sequence. Nucleotide and amino acid sequence of ginbuna CD3ε are shown. Predicted signal peptide, extracellular domain, transmembrane region, and cytoplasmic domain are labeled, CXXC motif and ITAM are boxed. Amino acid numbers are at right. (B) Schematic illustration of gCD3ε. Ginbuna CD3ε can be divided int...
Data
Specificity test of rabbit serum by immune-absorption. Absorption test of gCD3ε Ab was performed using transmembrane deletion mutant (TMDM) recombinant gCD3ε protein. Western blot analysis shows no band when anti-gCD3ε Ab was absorbed with antigen (TMDM rgCD3ε, right), while the Ab not absorbed with the antigen shows positive band (left).
Data
Migration of donor cells in recipient organs. CFSE-labeled donor cells were detected in recipient kidney, spleen, and peripheral blood leukocytes (PBL) on the histograms. Mean ± SD of more than three independent experiments are shown.
Data
CD3ε expression in tissues of other cyprinid species. Spleen and kidney leukocytes from carp (A) and goldfish (B) were stained with anti-CD4-1 and CD8α mAbs followed by Alexa Fluor® 488 anti-rat IgG, and stained with anti-gCD3ε Ab or anti-hZAP-70 mAb followed by 647 goat anti-rabbit IgG. Lymphocytes were gated on FS and SS dot plot. Mean ± SD of mo...
Article
The symbiosis between metazoans and prokaryotes is one of the most ancient and successful relationships found in nature. Mucosal surfaces are the main interface between animal hosts and their environment, and these surfaces are colonized by unique microbial communities that shape every physiological system of the host. Teleost fish have both an inn...
Article
We have monoclonal antibodies (mAbs) against CD4-1 (6D1) and CD8α (2C3) in ginbuna crucian carp Carassius auratus langsdorfii. In our previous studies we showed that 2C3 mAb positive cells are the primary cell type showing specific cytotoxicity against allogeneic targets, suggesting that CD8α⁺ lymphocytes in ginbuna are equivalent to cytotoxic T ly...
Article
Background/aim: Bacterial lipopolysaccharide (LPS) is involved in the activation of the innate immune responses on monocytes/macrophages in vitro, and by intravenous injection. Although small quantities of LPS are usually found in traditional Chinese medicines, vegetables and fruits, the mode of action of orally administered LPS is still unclear....
Article
Full-text available
Cartilaginous and bony fish are the most primitive vertebrates with a thymus, and possess T cells equivalent to those in mammals. There are a number of studies in fish demonstrating that the thymus is the essential organ for development of T lymphocytes from early thymocyte progenitors to functionally competent T cells. A high number of T cells in...
Article
Oral administration of lipopolysaccharide (LPS), a major outer-membrane component of Gram-negative bacteria, has been found to prevent infection in mammals and fish. Oral administration of LPS is believed to increase phagocytic activity and promote cytokine production, thus associating it with priming. The present study aimed to elucidate the effec...
Article
Full-text available
Neutral sphingomyelinase (nSMase) activation in response to environmental stress or inflammatory cytokine stimuli generates the second messenger ceramide, which mediates the stress-induced apoptosis. However, the signaling pathways and activation mechanism underlying this process have yet to be elucidated. Here we show that the phosphorylation of n...
Article
Granzymes are serine proteases released by cytoplasmic granules within cytotoxic T lymphocytes and natural killer (NK) cells. Granzymes induce apoptosis within virus‐infected and transformed cells. Information on the function of fish granzymes is limited, although granzyme‐like genes have been reported in several fishes. Here, we report the identif...
Article
The existence of fish-specific isoforms of interferon (IFN)c, known as IFNc-related (IFNcrel), has been reported in several fish species. However, comparisons with deduced amino acid sequences of known IFNcrels among several fish species have indicated significant differences at the C-terminus basic amino acid continuous sequences, which indicate t...
Article
The existence of fish-specific isoforms of interferon (IFN)γ, known as IFNγ-related (IFNγrel), has been reported in several fish species. However, comparisons with deduced amino acid sequences of known IFNγrels among several fish species have indicated significant differences at the C-terminus basic amino acid continuous sequences, which indicate t...
Article
Unlike mammals fish have two type II interferons, IFNγ and IFNγ‐related (IFNγrel). Both IFNγ and IFNγrel are structurally similar to human and mouse IFNγ and induce pro‐inflammatory responses. We previously identified two IFNγrel genes, IFNγrel 1 and IFNγrel 2 from ginbuna crucian carp, Carassius auratus langsdorfii . We produced recombinant IFNγre...
Article
Full-text available
Fish genomes possess three type II interferon (IFN) genes, ifnγ1, ifnγ2 and ifnγ-related (ifnγrel). The IFNγ-dependent STAT signalling pathway found in humans and mice had not been characterized in fish previously. To identify the antiviral functions and signalling pathways of the type II IFN system in fish, we purified the ifnγ1, ifnγ2 and ifnγrel...
Article
The presence of antigen-specific cytotoxic T cells has been suggested in a number of in vivo and in vitro studies in fish. Acute allograft rejection with an accelerated response on second-set grafts and the presence of graft-versus-host reaction (GVHR) has been reported in teleost. Alloantigen- and virus-specific cytotoxicity has also been demonstr...
Article
We have previously demonstrated the presence of graft-versus-host reaction (GVHR) in fish employing a model system of clonal triploid ginbuna and tetraploid ginbuna-goldfish hybrids. To elucidate the role of CD8alpha+ T cells in the induction of GVHR, we investigate the kinetics of CD4+ and CD8+ T-cell subsets in GVHR along with the pathological ch...
Article
CD8-positive (CD8(+)) cytotoxic T lymphocytes (CTL) have antigen-specific cytotoxic activity. In fish, however, CTL expressing CD8 on their cell surface have not been identified. In order to characterize the cells involved in specific cell-mediated cytotoxicity in teleosts, we separated and sorted ginbuna kidney leucocytes into CD8alpha(+), CD4(+)...

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