PROCEEDINGS
24th JAPANESE ASSOCIATION FOR
DEVELOPMENTAL & COMPARATIVE IMMUNOLOGY
Fukuoka Japan
July 9 to 14, 2012

日本比較免疫学会
第 24 回 学術集会講演要旨
会 期：２０１２年７月９日（月）～１３日（金）会 場： ヒルトン福岡シーホーク･ホテル
学 術 集 会 会 長：中尾学術集会 事務局長：杣本実樹（九州大学）
智軌（九州大学）

日本比較免疫学会―２０１２―

会場：ヒルトン福岡シーホーク

810-8650 福岡市中央区地行浜 2-2-3
TEL 0192-844-8111/FAX 092-844-7887

館内見取図The Symposium of
the 3 Japanese Societies;*Japanese Association for Developmental & Comparative Immunology (JADCI)*Japan Society for Comparative Endocrinology (JSCE)*Japanese Society for Comparative Physiology & Biochemistry (JSCPB)

Recognition Mechanisms-Current Topics in Comparative BiologyOrganized by Dr. Haruhisa Wago (Saitama Medical University)

July 9th, 2012 (PM 3:30-18:00)
at Hilton Fukuoka Sea Hawk Hotel, Fukuoka City

JADCI
Recognition of Pathogenic or Symbiotic Bacteria and
Immune Responses in Insects
Lee Bok LuelThe Global Research Lab (GRL) for Insect Symbiosis and National Research Laboratory (NRL) of
Host Defense Proteins, College of Pharmacy, Pusan National University, Jangjeon Dong, Kumjeong
Ku, Busan, 609-735, Korea, E-mail: brlee@pusan.ac.kr
The insect’s Toll receptor signaling pathway is responsible for the defense against
Gram-positive bacteria and fungi, while the immune deficiency (Imd) pathway is activated primarily
in response to Gram-negative bacteria. Both of these pathways lead to the expression of
antimicrobial peptides (AMPs) via the activation of NF-κB-like transcription factors. Even though
elegant Drosophila genetic studies characterized the innate immune responses in detail, there is some
limitation to reveal the biochemical mechanisms underlying invertebrate’s innate immune responses.
To investigate the biochemical basis of how insects recognize invading pathogenic microbes andhow these recognition signals are transferred to the down-stream factors and activate host innate
immune responses, it is necessary to use large insects, from which larger amounts of hemolymph(insect blood) can be extracted and can be used to perform biochemical studies. We have recently
determined molecular activation mechanisms of insect’s Toll signaling pathway after purification ofseveral pattern recognition proteins, serine proteases, serpins and AMPs. In this talk, we will discuss
the biochemical characterization of these molecules, which are involved in the recognition ofpathogenic microbes, in the amplification of recognition signal transduction and in the regulation ofproteolytic cascade leading to the production of AMPs in insects.
Also, I will discuss the molecular mechanisms of how insect hosts distinguish pathogenic
bacteria and symbiotic bacteria in order to defense their bodies in insects.

JADCI
Immunological Recognition in Starfish Larvae and Adults:
The Alteration of Immune Cells Via Metamorphosis
R. Furukawa1, 2
1

Department of Biology, Research and Education Center for Natural Sciences,
Keio University, Japan
2
School of Fundamental Science and Technology, Graduate School of Science and Technology,
Keio University, JapanThe starfish is a phylogenetically primitive animal, and hence provides us insight into theancestral immune system during evolution of the deuterostomes. I have been characterizing theimmune cells of the starfish larva, about which there has been little progress since the discovery ofphagocytosis by Metchnikoff. In these immune cells, there is the apparent replacement of the larvalmesenchyme cells with the adult coelomocytes via metamorphosis. In this symposium, I will focus
on the self/nonself recognition by both types of immune cell.
I have demonstrated that self recognition is strikingly altered in the two types of immune cell
via metamorphosis. The larval mesenchyme cells selectively phagocytose xenogeneic sperm, whenlive allogeneic and xenogeneic sperm are simultaneously injected into the body. This result
indicates that the larval mesenchyme cells recognize sperm of the same species as a self. In otherwords, they lack the allorecognition capability. Interestingly, the larval mesenchyme cells
phagocytose live allogeneic sperm treated with a mixture of various glycosidases. In the adult, theaggregate formation of coelomocytes occurs in response to allogeneic transplantation of
coelomocytes, but not in response to autotransplantation. This observation strongly suggests thatthe allorecognition capability is acquired after metamorphosis in the starfish immune system.
Recently, I have identified a novel opsonin against bacteria, ApSRCR1, as one of the nonself
recognition molecules. ApSRCR1 is expressed in both the larval mesenchyme cells and adultcoelomocytes. Significantly, ApSRCR1 differs in its activity as an opsonin between the two typesof cells. Although functional inhibition of ApSRCR1 leads to incomplete phagocytosis, thisinhibitory effect is much higher in the coelomocytes than in the mesenchyme cells. In addition,
biochemical analysis shows that ApSRCR1 exhibits not only higher expression, but also a higher
extent of modification by N-glycosylation in the coelomocytes than in the mesenchyme cells. Thus,
the importance of a certain type of immune molecule is altered in the immune system with alterationof the immune cells via metamorphosis. In addition to these findings, I will present a global
analysis comparing immune-related genes expression in the larval mesenchyme cells and the adultcoelomocytes.

JSCE
Comprehensive Analysis of Neuropeptides of Marine Invertebrates
S. Kato1, K. Ohno2, A. Fujiwara3, M. Awaji4, T. Matsumoto4, K. Yamano4,
M. Yoshikuni1
1

Department of Bioscience and Biotechnology, Kyushu University, Japan
2
National Institute for Basic Biology, USA
3
National Research Institute of Fisheries Science, JAPAN
4
National Research Institute of Aquaculture, JAPANIt has been emerging that neural systems of marine invertebrates produce a large number ofsecretable peptides which may play important roles in the control of various biological phenomenon.
This progress highly depends on the improvement of analytical instruments such as a DNA
sequencer and a mass spectrometer. Currently, we can identify a lot of peptides from neural tissuesof only a few individuals by using these updated instruments.
Comprehensive analysis of mRNAs provides a list of possible secretory peptides with aconfirmation of a signal sequence and convertase-cleaving sites. But, it will not give a list of
actually-secreted peptides. A mass spectrometer can directly detect small quantities of peptides
expressed in tissue samples. However, it will require repetitive laborious analyses for obtaining
accurate identification of peptides.
The combined use of mRNA analysis and mass spectrometry compensates imperfections of eachmethod to help rapid and accurate detections of secretory peptides. We comprehensively analyzed
ESTs and peptides of radial nerves of red sea urchin (Pseudocentrotus depressus) and visceralganglions of oyster (Crassostrea gigas). We have identified many peptides over a hundred including
large numbers of unknown sequences in each animal by the combined use of EST and mass
spectrometry. Some of those peptides were interestingly classified into several groups. These groups
are characterized with unique sequences of amino acids at the carboxyl-terminal ends, respectively.
Currently, the use of next generation DNA sequencer rapidly accelerates increase in number ofregistered sequence data at the SRA archive site of NCBI. The combined use of EST and MS will be
more powerful to identify new peptides and establish an information infrastructure forneuroendocrinology.

JSCE
Ligand recognition and signal transduction of GPCRs and Toll-like receptors of
the protochordate, Ciona intestinalis: what is evolutionarily common or unique?
H. Satake
Bioorganic Research Institute, Suntory Foundation for Life Sciences, Japan
Ascidians, invertebrate deuterostomes, belong to the subphylum Tunicata or Urochodata in thephylum Chordata. Their critical phylogenetic position as protochordates suggests that they conserve
evolutionary origins of various biological systems of vertebrates: nervous, neuroendocrine, endocrine
and innate immune systems. Over the past few years, the cosmopolitan species, Ciona intestinalis,has been shown to possess orthologs or prototypes of vertebrate neuropeptides, peptide hormones,
and their receptors including gonadotropin-releasing hormones (GnRHs) and their receptors. To date,seven GnRH family peptides (t-GnRH3 to -8 plus Ci-GnRH-X) and four their receptors(Ci-GnRHR1 to -4) have been identified in C. intestinalis. t-GnRHs, like vertebrate counterparts,
activate Ci-GnRHRs in a ligand-selective manners, whereas Ci-GnRH-X moderately inhibits theactivities of t-GnRHs at Ci-GnRHR1 and -3. A striking feature of t-GnRH signaling is that the
Ciona-specific GnRHR orphan paralog, Ci-GnRHR4, heterodimerizes with Ci-GnRHR1 and -2,
leading to the differential regulation of signal transduction via the two receptors without ligand
binding affinity altered. These findings reveal that C. intestinalis, which lacks a pituitary (a major
target of hypothalamic GnRH in vertebrates), conserves GnRH signaling essentially common tovertebrates and established the species-specific signaling regulatory mechanisms.
C. intestinalis has also been found to possess Toll-like receptors (TLRs) that play crucial roles ininnate immunity of vertebrates. Intriguingly, only two TLR genes are present in the C. intestinalis
genome, while other deuterostome invertebrates, sea urchin and amphioxus harbor 222 and 72 TLR
candidates. Of particular interest is that the two C. intestinalis TLRs, i.e., Ci-TLR1 and -2, activate
NF-B in response to multiple TLR ligands which are recognized by different mammalian TLRs:
zymosan for human TLR2 (hTLR2), heat-killed Legionella pneumophila for hTLR2, and poly(I:C)
for hTLR3. These data verify both common and unique functionality of Ci-TLRs, compared withmammalian orthologs. In addition, such recognition by Ci-TLRs cannot be predicted by sequencehomology-based search, suggesting the molecular diversity in ligand recognition by non-vertebrate
TLRs. Collectively, species-specific multiplication and loss of genes for endogenous ligands andreceptors, including peptides, GPCRs, and TLRs, generate various and unique molecular recognition,
which is likely to be implicated in the evolution and diversity of organisms.

JSCPB
OPN5, a Photosensory Protein for Mammalian Ultraviolet Photoreception
D. Kojima
Department of Biophysics and Biochemistry, Graduate School of Science,
The University of Tokyo; PRESTO, Japan Science and Technology Agency, JapanThe ultraviolet (UV) component of sunlight is utilized in a variety of animal species fortheir environmental cues, e.g., for flower discrimination and orientation/navigation in insects and formate choice and parent-offspring communication in birds: Consistently, these animals have
UV-sensitive photoreceptors in their eyes. In mammals, some rodents such as mice have
UV-sensitive retinal cone photoreceptor cells, whose function remains yet unknown, while mostother mammalian species including human beings have been believed to lack such UV
photoreception mechanisms. Here I report that mammalian OPN5 protein (neuropsin) is a
UV-sensitive photoreceptor. The Opn5 gene encoding OPN5 protein has been first identified in themouse and human genomes as a member of rhodopsin (opsin) family, most of which encode
photosensitive proteins. The mouse Opn5 mRNA expression is detected in various neural tissues,
though its detailed cellular expression pattern has not been reported. Furthermore, it has remained
unclear whether OPN5 functions as a GPCR-type photopigment or photoisomerase, or has functions
other than those. This study was aimed to examine molecular function of mammalian OPN5 and its
detailed expression pattern in the tissues.
Mouse OPN5 protein bound a chromophore 11-cis-retinal and exhibited an absorption
maximum at 380 nm. Upon UV-light illumination, OPN5 was converted to a blue-absorbing
photoproduct, which was stable in the dark and reverted to the UV-absorbing state by the subsequentorange light illumination, indicating its bistable nature. Human OPN5 also had the absorption
maximum at 380 nm with spectral properties similar to mouse OPN5, revealing that OPN5 is the firstand hitherto unknown UV light-sensitive opsin in human beings. OPN5 had the ability to activate
hetrotrimeric G-protein Gi in a UV-dependent manner. Immuno-blotting analyses of mouse tissue
extracts identified retina, brain and, unexpectedly, outer ear as the major sites of OPN5 expression.
In the mouse tissue sections, OPN5 were detected in a subset of non-rod, non-cone retinal neurons aswell as in the epidermal and muscle cells of the outer ear. Most of these OPN5 immuno-positive
signals were co-localized with the Gi-positive signals. These results demonstrate the first example of
UV-sensitive photoreceptor in human beings and strongly suggest that OPN5 transduces
UV-sensitive Gi-mediated signaling pathway in the mammalian tissues.

JSCPB
Specificity and Sensitivity of Sex Pheromone Perception
in the Silkmoth Bombyx moriT. Sakurai, M. Tabuchi, R. Kanzaki
Research Center for Advanced Science and Technology, The University of Tokyo, Japan
Male moths locate their mates using species-specific sex pheromones emitted by conspecificfemales. One striking feature of sex pheromone recognition in male moths is the high degree of
specificity and sensitivity at all levels from primary sensory processes to behavior. Despite advancesin our understanding of molecular, structural, and physiological mechanisms underlying pheromone
detection at peripheral level, mechanisms that define behavioral response specificity to conspecificpheromones and the ability to display behavioral responses to minuscule amounts of pheromone
molecules are still obscure. In this talk, recent work that addresses these two questions using a
genetically tractable moth species the silkmoth, Bombyx mori, will be presented.
In the silkmoth a single pheromone component, (E,Z)-10,12-hexadecadienol (bombykol), issufficient to elicit full sexual behavior in males. The sex pheromone receptor BmOR1 expressed in
specialized olfactory receptor neurons (ORNs) mediates specific detection of bombykol in theantennae of male silkmoths. To examine causal relationships between sex pheromone receptor
specificity and behavioral specificity, we genetically expressed the sex pheromone receptor PxOR1of the diamondback moth Plutella xylostella in BmOR1-expressing ORNs. Expression of PxOR1
conferred the ability to respond to its specific ligand, (Z)-11-hexadecenal, at electrophysiological andbehavioral levels. These results indicate that activation of BmOR1-expressing ORNs alone issufficient to trigger full sexual behavior. Thus, behavioral specificity is determined by sex
pheromone receptor ligand specificity in the silkmoth.
We also examined how the sensitivity of ORNs is processed in the brain to generate highbehavioral responsiveness. To precisely control the activity of BmOR1-expressing ORNs, we
genetically introduced a light-gated ion channel, channelrhodopsin-2, in these ORNs. Usingpaired-pulse photostimulation at various interstimulus intervals, we found that ORN activity that is
subthreshold in terms of behavior can be amplified to suprathreshold levels by temporal integrationin antennal lobe projection neurons (PNs) if occurring within a specific time window. Temporal
integration in PNs was only observed for weak inputs but not for strong inputs. These results showthat temporal integration of ORN inputs in PNs contributes to high behavioral sensitivity by lowering
the threshold of behavioral responsiveness.

Oral presentation

Session: Anti viral immunity
Functions of CD8-positive and CD4-positive lymphocytes against virus-infection
in ginbuna crucain carp
Tomonori Somamoto1, Teruyuki Nakanishi2, Miki Nakao1
1

Department of Bioscience and Biotechnology, Kyushu University, Japan
2
College of Bioresource Sciences, Nihon University, Japan

Recent studies have shown that cytotoxic and helper T-cell functions are well-conserved in teleostfish. However, information on their functions against virus-infection is still limited. The present
study demonstrated the functions of CD4-positive and CD8-positive lymphocytes employing clonal
ginbuan crucian carp, anti-ginbuna CD8α and CD4 mAb and crucian haematopitetic necrosis virus(CHNV). The cytotoxic assays using syngeneic cell line as a target showed that CD8-positive
lymphocytes from CHNV-sensitized fish are significantly capable of killing infected-syngeneic
target cells. The inhibition of the activity by concanamycin A suggests that virus-specific CTL killvirus-infected cells utilizing perforin/granzyme pathway. However, the activity of CD8-positive cells
is comparable or lower than that of monocytes-enriched effector cells or CD8-negative lymphocytes.
This finding suggests that CTLs may be not dominant effector cells that are induced by
CHNV-infection. To confirm whether helper T-cells (Th cells) of teleost fish actually facilitate
secondary humoral and cellular immune responses, the adoptive transfer study was performed using
clonal donors and recipients. Naïve recipients receiving CHNV-sensitized donor cells started to
produce specific antibody at 5 days post infection (dpi), while antibody production in the recipients
receiving non-sensitized cells or CHNV-sensitized Th cell-depleted cells became detectable at 12
dpi. This finding suggests that the adoptive transfer of Th cells provide the recipient faster antibody
response similar to secondary ones. On the other hand, although adoptive transfer of sensitized donor
cells into naïve fish induced a significant enhancement of cell-mediated immunity, no significant
difference of the cell-mediated immune response was observed in the recipients receiving thesensitized donor cells with or without Th cells. These results indicate that Th cells of teleost fish canhelp B-cells, but it remains unclear whether they can help CTLs. Thus, further study would beneeded to understand the induction system of humoral and cell-mediated immunity in teleost fish.

Session: Complement and Complement-like factors
Cytolytic factor in the mosquito

Toshinori Sasaki, Mutsuo Kobayashi

Department of Medical Entomology, National Institute of Infectious Diseases,
Shinjuku-ku, Tokyo, Japan, 162-8640

Although cytolysis of invading organisms is an innate form of immunity used by invertebrates, so far
the underlying mechanism remains less explored. The pupal hemolymph of the mosquito Armigeres
subalbatus induces an activity that causes hemolysis of human red blood cells (HRBC). This
hemolytic activity was inhibited by sialic acid (N-acetylneuraminic acid) and serine proteaseinhibitors. We purified the sialic acid-specific lectin(s) from the pupal hemolymph using
formaldehyde-fixed HRBC and determined the sequence of the amino-terminal 19 amino acidresidues. A polyclonal antibody produced against this N-terminal peptide clearly inhibited thehemolytic activity of the hemolymph in vitro, thus suggesting that the hemolysis of HRBC is causedby the lectin present in the mosquito hemolymph. We suggest that mosquitoes possess a cytolysis
system.

Isotypic Diversity in the Ontogenetic Expression of the Complement Component
in the Common Carp (Cyprinus carpio)
Vo Kha Tam1, Chie Okura1, Masakazu Kondo2, Tomonori Somamoto1, Miki Nakao1
1

2

Department of Bioscience and Biotechnology, Kyushu University, Japan.
Department of Applied Aquabiology, National Fisheries University, Shimonoseki 759-6595, Japan.

Isotypic diversity of complement components is a striking feature of teleost complement system. Asa first line of innate defense, the complement system has been considered as an important clue ofhumoral defense in early development, however functional diversity of the isotypes during teleostontogeny is poorly understood. The present study aimed at clarifying comprehensive picture of
ontogenetic expression of the diversified complement component isotypes in carp. Real-timequantitative PCR detected embryonic expression of C1r/s, MASPs, factor B/C2, C3, C4, C5, C6, C7,
C8, C9, and factor I. A remarkable difference in the expression time course was noted between theisotypes in C3, C4, and C5. Especially, teleost-specific isotypes of C3 and C4 (non-histidine-type)started around hatching, in contrast to evolutionarily common isotypes (histidine-type), whichshowed much earlier expression. Whole-mount in situ hybridization of carp embryos revealed somedifference in embryonic expression sites of two major C3 isotypes (C3-H1 and C3-S) in addition tocommon expression sites such as the yolk syncytial layer. The temporal and spatial differences in
expression among the isotypes suggest that the isotypes are functionally differentiated in teleost early
development.

Identification of Pattern-Recognition Molecule in Hagfish Complement System
Tomokazu Yamaguchi1, Kazufumi Takamune1, Masakazu Kondo2,
Yukinori Takahashi2,Yoko Kato-Unoki3, Miki Nakao4, Tamotsu Fujii5
1

Graduate School of Science and Technology, Kumamoto University, Japan
Department of Applied Aquabiology, National Fisheries University, japan
3
Faculty of Agriculture, Kyushu University, japan
4
Department of Bioscience and Biotechnology, Kyushu University, japan
5
Department of Health Sciences, Hiroshima Prefectural University, japan

2

All extant jawed vertebrates share a common adaptive immune system in which immunoglobulin
domain-based molecules act as antigen receptors. On the other hand, jawless vertebrates, lamprey
and hagfish, use variable lymphocyte receptors composed of leucine-rich repeat cassettes for antigen
recognition. Since invertebrates and primitive chordates do not have adaptive immune system,
immune system seems to change dramatically in course of evolution from primitive chordate tognathostome. From a phylogenic perspective of defense mechanisms, previously we foundcomplement C3 and mannose-binding lectin-associated serine protease 1 (MASP-1) in hagfish andsuggested the involvement of lectin pathway in hagfish innate immune system. In this study, we
focused on the pattern recognition molecules in the hagfish, Eptatretus burgeri, and tried to purify itfrom serum by affinity chromatography. When the serum was treated with GlcNAc-agarose,
followed by successive elution of the binding molecules with GlcNAc and EDTA, mainly four
proteins (31 kDa, 27 kDa, 26 kDa, and 19 kDa) and 26 kDa protein were detected in the
GlcNAc-eluate and EDTA-eluate, respectively. Collagenase treatment showed the presence ofcollagen-like domain only in the 26 kDa protein in the EDTA-eluate. Since common patternrecognition molecules such as mannose-binding lectin possess collagen-like domain, we examinedthe entity of the 26 kDa protein by sequencing its N-terminal amino acids and cDNA obtained by 3’
and 5’ RACE methods, and identified it as a member of C1q family. Herein, it will be referred to as
hagfish C1q (hagC1q). Western blot analyses using anti-hagC1q, MASP-1, and complement C3
antibodies showed that hagC1q associated with MASP-1 and complement C3 in the serum and hadbinding ability to Escherichia coli as a divalent cation-dependent manner. These results suggest thathagC1q plays an important role in hagfish innate immune system.

Microbe-specific C3b deposition in the horseshoe crab complement system
in a C2/factor B-dependent or -independent manner
K. Tagawa1, T. Yoshihara1, T. Shibata2, K. Kitazaki1, Y. Endo3, T. Fujita3,
T. Koshiba1,2, S. Kawabata 1,2
1

Graduate School of Systems Life Sciences, Kyushu University, Japan
Department of Biology, Faculty of Sciences, Kyushu University, Japan3
Department of Immunology, Fukushima Medical University School of Medicine, Japan
2

Complement C3 plays an essential role in the opsonization of pathogens in the mammaliancomplement system, whereas the molecular mechanism underlying C3 activation in invertebrates
remains unknown. To understand the molecular mechanism of C3b deposition on microbes, wecharacterized two types of C2/factor B homologs (designated TtC2/Bf-1 and TtC2/Bf-2) identified
from the horseshoe crab Tachypleus tridentatus. Although the domain architectures of TtC2/Bf-1 and
TtC2/Bf-2 were identical to those of mammalian homologs, they contained five-repeated andseven-repeated complement control protein domains at their N-terminal regions, respectively.
TtC2/Bf-1 and TtC2/Bf-2 were synthesized and glycosylated in hemocytes and secreted tohemolymph plasma, which existed in a complex with C3 (TtC3), and their activation by microbes
was absolutely Mg2+-dependent. Flow cytometric analysis revealed that TtC3b deposition was
Mg2+-dependent on Gram-positive bacteria or fungi, but not on Gram-negative bacteria. Moreover,this analysis demonstrated that Ca2+-dependent lectins (C-reactive protein-1 and tachylectin-5A)
were required for TtC3b deposition on Gram-positive bacteria, and that a Ca2+-independent lectin(Tachypleus plasma lectin-1) was definitely indispensable for TtC3b deposition on fungi. In contrast,
a horseshoe crab lipopolysaccharide-sensitive protease factor C was necessary and sufficient todeposit TtC3b on Gram-negative bacteria. We conclude that plasma lectins and factor C play key
roles in microbe-specific TtC3b deposition in a C2/factor B-dependent or -independent manner.

Session: Cytokine and Chemokine
A chemokine expressed in the secondary lymphoid organs
in of fugu (Takifugu rubripes)
H. Tauchi1, H. Suetake1, 2, T. Odaka2, M. Kaneda2, T. Maeda2, K. Kikuchi1,
Y. Suzuki1, T. Miyadai2
1Fisheries Laboratory, Graduate School of Agricultural and Life Sciences,
The University of Tokyo, Japan
2
Faculty of Marine Bioscience, Fukui Prefectural University, Japansecondary lymphoid organs differ substantially from those of mammals. They are composed of thespleen and kidney and lack lymph nodes and germinal centers. Understanding adaptive immune
responses in fish requires detailed characterization of the microarchitecture of their secondary
lymphoid organs. Lymphoid chemokines such as CCL19 and CCL21 regulate trafficking oflymphocytes and antigen-presenting cells to secondary lymphoid organs, facilitating adaptive
immune responses. In the present study, we surveyed the fugu genomic database and found 3 CCL19
or CCL21-like genes. Two of the 3 genes encode a CC chemokine resembling CCL19, with 4
cysteines, and they are evidently duplicated genes, as there is only a single nucleotide difference
within their coding regions. The third gene encodes a CC chemokine including 6 cysteines such as
mammalian CCL21. This chemokine was expressed in the spleen and kidney, whereas the other 2
were not expressed in these organs. This indicates that this chemokine is the secondary lymphoidchemokine in fish. In situ hybridization studies showed that this lymphoid chemokine was expressedin melanomacrophage centers (MMCs). These results suggest that fish MMCs play pivotal roles in
adaptive immune responses.

Identification, characterization and expression analysis of IL-17 receptors, Act1and TRAF6 genes in Japanese pufferfish (Takifugu rubripes)
H. Korenaga1, T. Kono2 , M. Sakai1
1Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Japan
2
Interdisciplinary Research Organization, University of Miyazaki, Japan

Interleukin-17 (IL-17) is the novel pro-inﬂammatory cytokine family with six members (A-F) thatdeﬁnes a new class of CD4effector T cells, termed as Th17. Teleost IL-17 was reported to consist ofseven ligands including teleost specific member, IL-17N in some species. However, IL-17 receptors
and their signaling are few reported in teleosts. In mammals, IL-17 receptors (5 members, RA-RE)
have a cytoplasmic tail with motifs similar to the TIR superfamily, now termed SEF/IL-17R(SEFIR) domain. Act1 interacts with the IL-17 receptors through the C-terminal SEFIR domain. In
addition to interacting with the SEFIR domains of IL-17 receptors, Act1 also functions as an adaptor
protein by binding to additional signaling molecules, most notably recruitment of TNF receptor
associated factor 6 (TRAF6) by two putative TRAF-binding sites. This signaling activates the NF-kB
transcription factor pathway. In this report, we identified seven IL-17 receptors (RA, RC-1, -2, RD-1,-2, -3 and RE), Act1 and TRAF6 from Japanese pufferfish, Takifugu rubripes using comparative
genomics based on chromosome synteny and characterized their biological activities. Fugu
IL-17RC1 and RE genes harbored on scaffold_180 and other Fugu IL-17 receptors were found to belocated on independent contigs. Fugu IL-17 receptors showed single transmembrane
domain-containing receptors, ranging in size from 470 to 755 amino acid. Moreover, these receptors
contained certain conserved the SEFIR domain. Fugu Act1 contained TRAF binding sites at the
N-terminus, and Act1 had a SEFIR domain located at the C-terminus. Fugu TRAF6 ORF is
predicted to encode 561 amino acid. Multiple alignment with other known TRAF6 genes revealed
conservation among vertebrates. The expression pattern of Fugu IL-17 RA and RC genes wereobserved in almost all organs, while the other Fugu IL-17 receptor genes expressed in few organs. In
particular, IL-17 receptor genes showed highly expression in the spleen. Moreover, expression of
IL-17 receptor genes showed different pattern in the mitogen-stimulated head kidney cells. In
addition, expression analysis of Fugu Act1 and TRAF6 genes may play important roles in
inflammatory responses in fish.

Session:Ig and other antigen receptors
VLRC defines a population of lamprey lymphocytes distinct from that expressing
VLRA or VLRB
Yoichi Sutoh, Jun Kasamatsu, Noriyuki Otsuka, Yukiko Miyatake, Masanori Kasahara
Department of Pathology, Hokkaido University Graduate School of Medicine,
Sapporo 060-8638, Japan
Instead of T-cell and B-cell receptors (TCR and BCR), jawless vertebrates such as lamprey andhagfish use variable lymphocyte receptors (VLRs) as antigen receptors. VLRs generate diversity
comparable to that of TCR or BCR by assembling highly diverse leucine-rich repeat (LRR) modules
presumably via cytidine deaminases. Like TCR and BCR, they appear to exhibit allelic exclusion
with each lymphocyte expressing a unique VLR molecule. We have previously shown that VLR
monomers adopt a horseshoe-shaped structure characteristic of LRR family proteins and that
hypervariable residues are located predominantly on the concave surface, suggesting that VLR binds
antigen on its concave surface. This suggestion was confirmed by the crystallographic analysis oflamprey VLR molecules in complex with the H-trisaccharide from human blood type O erythrocytes.
Both hagfish and lampreys have two types of VLRs, named VLRA and VLRB. VLRB occurs as
pentamers or tetramers of dimers and constitutes major agglutinating antibodies produced in
response to antigen stimulation. On the other hand, VLRA is a membrane-bound receptor notsecreted into the serum. Recent evidence indicates that lamprey VLRA+ and VLRB+ cells display
gene expression profiles resembling those of T- and B-cells of jawed vertebrates, respectively,
suggesting that lampreys also have a dual antigen receptor system analogous to that of jawedvertebrates. Here, we describe the identification of a third lymphocyte lineage expressing a distinct
antigen receptor, VLRC. Single cell genomic PCR indicates that VLRC is assembled in a populationof lymphocytes distinct from VLRA+ or VLRB+ cells. The apparent presence of a transmembraneregion and the failure to detect a secreted form indicate that VLRC is a membrane-bound antigen
receptor. Thus, like jawed vertebrates that have B-cells and two major populations of T-cells ( andT cells), lampreys appear to have three lineages of lymphocyte-like cells, of which one is
B-cell-like and two are presumably T-cell-like.

Session: Innate Cellular immunity
Phagocytosis and bactericidal abilities of teleost thrombocytes
Takahiro Nagasawa1, Chihaya Nakayasu2, Tomomasa Matsuyama2, Aja M. Rieger3,
Daniel R. Barreda3, Tomonori Somamoto1, Miki Nakao1
1

2

Department of Bioscience and Biotechnology, Kyushu University, Japan
National Research Institute of Aquaculture, Fisheries Research Agency, Japan3
Department of Biological Sciences, University of Alberta, Canada

Thrombocytes have been recognized as hemostatic cells in nonmammalian vertebrates. Unlike
mammalian platelets, thrombocytes are nucleated cells with a lymphocyte-like morphologicalfeature, and possible involvement of thrombocytes in innate immune function has been considered inaddition to the hemostatic function. In the present study, we report phagocytic abilities of someteleost thrombocytes. Using a monoclonal antibody specific for thrombocytes of common carp
(Cyprinus carpio), thrombocytes were isolated from peripheral blood and examined for expression ofvarious immune-related genes, resulting in detection of significant level of lysozyme, iNOS andMHC class II using RT-PCR. Upon flow cytometry-based phagocytosis assay, a number ofthrombocytes ingested fluorescent latex particles (0.5 μm, 1 μm, 2 μm and 3 μm), bacteria(Escherichia coli) and zymosan particles. Phagocytosis by the thrombocytes was also confirmed byfluorescent microscopy and transmission electron microscopy, which revealed internalization ofthese particles into thrombocytes. We also observed that thrombocytes of the olive flounder(Paralichthys olivaceus) had similar phagocytic behavior. These data indicate that thrombocytes of
those species are potent phagocytes, suggesting that those phagocytic characteristics of thrombocytes
are widely conserved in teleosts. We also assessed a phagolysosome formation ability of teleostthrombocytes against the ingested pathogens, for detecting intracellular bactericidal activities of
those thrombocytes. By using an ImageStream multi-spectral flow cytometer, we assessedphagolysosome fusion of goldfish (Carassius auratus) thrombocytes. On this analysis we detectedthat lysosomes of these thrombocytes visualized with fluorescent dextran were colocalized theingested small beads and were formed a ring around a large beads like other typical phagocytes.
Overall, the results indicate that teleost thrombocytes have dual functions as not only hemostatic
cells but also as phagocytic immune cells against microbial infections.

Session: Invertebrate immune responce
Transglutaminase-catalyzed Relish Crosslinking Suppresses Innate Immune
Signaling in the Drosophila Gut

T. Shibata1, S. Sekihara2, T. Fujikawa2, R. Miyaji2, T. Ishihara1,2, T. Koshiba1,2,
S. Kawabata1,2
1Department of Biology, Faculty of Sciences, Kyushu University, Fukuoka, Japan.
Graduate School of Systems Life Sciences, Kyushu University, Fukuoka, Japan.

2

Mammalian transglutaminases (TGs) play important roles in numerous physiological phenomena
such as blood coagulation and skin formation via protein-protein crosslinking. We show that
Drosophila cytoplasmic TG suppresses innate immune signaling in the gut. TG-RNA-mediatedinterference (TG-RNAi) caused a short life span under non-sterile conventionally reared conditions,but not under germ-free conditions. Under conventionally reared conditions, TG-RNAi enhanced theexpression of immune deficiency (IMD) pathway-controlled antimicrobial peptide genes. Ingestionof gut lysates prepared from conventionally reared TG-RNAi flies into non-TG-RNAi flies resulted
in the short life span of the recipients. TG-RNAi under conventionally reared conditions triggered
severe apoptosis in the gut and induced the translocation of Relish, the NF-κB-like transcription
factor of the IMD pathway, from the cytoplasm to the nucleus. Ingestion of synthetic amine donors
by non-TG-RNAi flies, which inhibits the TG-catalyzed protein-protein crosslinking reaction, also
induced the nuclear translocation of Relish and enhanced the expression of IMD-controlled
antimicrobial peptide genes in the gut. We conclude that TG-catalyzed Relish crosslinking
suppresses the IMD-signaling pathway to maintain a buffered threshold required for immune
tolerance against commensal microbes.

Session: T-cell, MHC, and Ag-presentation
Long-term Transspecies Dimorphism of the PSMB8 Genein the Actinopterygian MHC Region
Naoko T Fujito1, Kentaro Tsukamoto2, Chris T Amemiya3, Masaru Nonaka1
1

Department of Biological Sciences, Graduate School of Science, University of Tokyo, Japan
2
Institute for Comprehensive Medical Science, Fujita Health University, Japan
3
Benaroya Research Institute, Virginia Mason Medical Center, USA

The proteasome subunit beta type-8 (PSMB8) gene encodes a catalytic subunit of theimmunoproteasome, which is involved in the generation of peptides presented by MHC class I
molecules. Two highly divergent types of PSMB8 termed the A and F types, possessing Ala or Val
and Phe or Tyr, respectively, at the 31st residue of the mature peptide located in the S1 pocket, havebeen identified in amphibian Xenopus species, teleost species belonging to Cypriniformes(zebrafish), Salmoniformes (trout), Beloniformes (Oryzias species), and Tetradontiformes(pufferfish), and elasmobranch sharks. Phylogenetic analysis indicated the presence of two veryancient lineages, the PSMB8F and PSMB8A lineages. The F type PSMB8 genes of Salmoniformes,
Cypriniformes and sharks belong to the PSMB8F lineage, whereas all the other sequences belong to
the PSMB8A lineage, showing that these two lineages have been established before the divergence of
sharks from the bony vertebrates and that the PSMB8F lineage has been lost in higher teleosts andtetrapods. However, in Xenopus and higher actinopterygian teleosts (Oryzias and pufferfish), the new
F type genes, possible functional equivalents of the PSMB8F, have revived as alleles within the
PSMB8A lineage independently. To clarify the evolutionary history of the PSMB8A and PSMB8F
lineage genes, we identified both of them in bichir, Polypterus senegalus, one of the most basal
actinopterygian. Segregation analysis showed that the two lineage genes are alleles in bichir as wellas in zebrafish and trout, whereas those of sharks are predicted to be paralogs behaving as
pseudoalleles. These results suggested that the highly divergent allelic lineages of PSMB8 have beenretained for more than 400 million years in lower actinopterygians by an extremely long-termbalancing selection. To clarify the genomic background of the PSMB8 dimorphism we also
performed a physical analysis of the MHC region around the bichir PSMB8 gene, using a bacterial
artificial chromosome (BAC) clone. The order and orientation of the flanking genes of PSMB8 ofbichir was identical with that of X. tropicalis but quite different from those of higher teleosts,
indicating that extensive genomic reorganization of the MHC region occurred in the teleost lineage
after the divergence of bichir.

Evolution of Teleost MHC Class II Genes
Revealed by Comprehensive Analysis of Medaka
H. Bannai1, M. Nonaka1
1

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, JapanThe Major Histocompatibility Complex (MHC) class II molecules play central roles in acquired
immunity by regulationg immune response through activation of CD4 T cells. Each individual
usually possesses one to a few classical MHC class II genes, which encode the MHC class II
molecules, and some non-classical MHC class II genes, which encode other than MHC class II
molecules. A full complement of the MHC class II genes has been elucidated only in mammalian
species thus far. In mammals the lineage of the MHC class II genes have been conserved, although
the data is incomplete and its evolution is still uncertain for non-mammalian vertebrates. To
understand evolution of the MHC class II genes, we performed the first comprehensive analysis of
the MHC class II genes in poikilothermic species using a teleost, medaka (Oryzias latipes). Throughdatabase search, cDNA cloning and genomic PCR, medaka was shown to possess five pairs ofexpressed class II genes, comprising one IIA and one IIB genes. Each pair is on a different
chromosome and not linked to the class I genes. Only one pair showed a high degree ofpolymorphism and was considered to be classical class II genes, whereas the other four pairs werenon-classical. Upon phylogenetic analysis of all medaka class II genes and most reported teleostclass II genes, IIA and IIB genes formed separate clades, each containing three well-corresponding
lineages. One lineage contained three medaka genes and all known classical class II genes ofOstariophysi and Euteleostei. The other two lineages contained one non-classical medaka gene eachand some Euteleostei genes. These results indicate the presence of multiple lineages of the teleost
MHC class II genes which have been conserved over three hundred million years.

Clonal culture of carp Th2-like cells whose immune status is hallmarked
by IL-4/13B gene expression
Takuya Yamaguchi1, Fumihiko Katakura1, Kazue Someya1, Takeshi Yabu1,
Johannes M. Dijkstra2, Tadaaki Moritomo1, Teruyuki Nakanishi1
1

Laboratory of Fish Pathology, Department of Veterinary Medicine,
Nihon University, JAPAN2
Institute for Comprehensive Medical Science, Fujita Health University, JAPAN
CD4+ T helper (Th) cells play a central role in modulating vertebrate immune responses. In
mammals, Th cells are subdivided into Th1, Th2, Th17 and Treg, according to their cytokine
production profiles and expression of lineage-specific transcription factors. However in teleosts,
many Th-related molecules have been identified but Th subsets have not been identified.
Identification of Th subsets will help to understand the teleost immune system. Recently, wedeveloped a bulk culture system in which co-culturing carp (Cyprinus carpio) leukocytes withsupporting cells resulted in long-term proliferation of CD4+ αβT cells. To characterize thesebulk-cultured T cells by their cytokine production profile, we stimulated them withphytohaemagglutinin (PHA) and examined the expression of Th-related genes. Expression of both a
Th1-related cytokine (IFNγ) and Th2-related cytokines (IL-4/13A and IL-4/13B) were enhanced by
PHA stimulation. In addition, bulk-cultured T cells expressed transcripts of the Th1 and Th2 master
regulators (t-bet and gata3, respectively). These results suggest that bulk-cultured T cells contained
both Th1- and Th2-like cells. We attempted to identify these Th subsets by establishing andanalyzing T cell clones. To establish T cell clones, single cells were picked up from bulk-cultured T
cells, seeded onto the supporting cells and cultured. Cells proliferated in 1–4 wells of each 96-wellplate. The majority of the clones expressed both TCR-α and -β, and also expressed either TCR-γ or-δ. Therefore these clones could produce the TCRαβ heterodimer but could not produce the TCRγδ
heterodimer. These clones also expressed the TCR co-receptor gene CD4-1, but not CD4-2, CD8α orCD8β. In addition, gata3 was expressed whereas t-bet was not. The gene expression profiles indicatethat these clones also have characteristics consistent with CD4+ αβT cell identity. One of theseclones (KoThL5) continued to proliferate and was successively transferred for more than 10 months
and 90 – 100 passages. In KoThL5 cells, PHA stimulation significantly enhanced the expression of
IL-4/13B, but did not enhance the expression of IFNγ or another Th2-related cytokine, IL-4/13A.
Together, these results indicate that KoThL5 cells represent a Th2-like subset which is hallmarked
by enhanced expression of IL-4/13B and gata3 expression.

Session: Vertebrate adaptive immune responce
Cell-mediated and humoral immune response to Edwardsiella tarda infection
in ginbuna crucian carp, Carassius auratus langsdorfii
M. Yamasaki1, K. Araki2, T. Nakanishi3, C. Nakayasu4, A. Yamamoto2
1

The United Graduate School of Agricultural Sciences, Kagoshima University, Japan
2
Faculty of Fisheries, Kagoshima University, Japan
3
College of Bioresource Sciences, Nihon University, Japan
4
National Research Institute of Aquaculture, Fisheries Research Agency, Japan

Edwardsiella tarda is an intracellular pathogen, causes edwardsiellosis in fish. Cell-mediatedimmunity (CMI) has a major role in the protection against intracellular bacterial infection in
mammals. In contrast the principal immune system against E. tarda in fish is still unclear. In order toclarify the importance of CMI and/or humoral immunity to prevent E. tarda infection in fish, we
examined elimination of bacteria, CMI-related gene expression, population of leukocytes andproduction of antigen-specific antibodies in E. tarda infected ginbuna crucian carp. Ginbuna crucian
carp (Carassius auratus langsdorfii) were infected with E. tarda at 105 CFU/100 g body weight (0.2
LD50) by intraperitoneal injection and tissue (kidney, spleen and liver), kidney leukocytes (KLs) andplasma were collected 0, 2, 4, 8, 12, 16, 30 days post-infection. Bacterial counts in the tissues were
determined by miles and misra methods. The expression analysis of CMI related genes such as IFNγ
and perforin in KLs was performed by semi-quantitative RT-PCR. Proportion of CD8α+ and sIgM+
cells in KLs were examined by influences assay with monoclonal antibody against ginbuna CD8α
and IgM, respectively. E. tarda-specific antibody titer in plasma was determined by ELISA.
Expression level of IFNγ and perforin were up-regulated from 2 days to 12 days post-infection. In
addition proportion of CD8α+ cells was increased from 4 days to 8 days. These results suggested that
CMI was induced in fish infected with E. tarda. The reduction of bacterial counts was observed from
2 days to 12 days, suggesting that CMI mainly contributed to elimination of E. tarda in the earlystage of E. tarda-infection. On the other hand, E. tarda-specific antibody titer was increased after
clearance of bacteria, indicating that protection against E. tarda-infection should be able to occur
without humoral immunity. These findings suggested that CMI might be the principal immune
system in the protection against E. tarda in fish.

Protection of ginbuna crucian carp (Carassius auratus langsdorfii) against
Edwardsiella tarda infection using adoptive transfer of cytotoxic T lymphocytes
K. Araki1, M. Yamasaki2, T. Nakanishi3, C. Nakayasu4, A. Yamamoto1
1

2Faculty of Fisheries, Kagoshima University, JapanThe United Graduate School of Agricultural Sciences, Kagoshima University, Japan
3
College of Bioresource Science, Nihon University, Japan
4
National Research Institute of Aquaculture, Fisheries Research Agency, Japan

Cell-mediated immunity (CMI) plays a major role in protection against intracellular pathogens inmammals. In particular, cytotoxic T lymphocytes (CTLs) play a crucial role in resolving intracellular
infections. Activated CTLs induce effector molecule-mediated lysis of the infected cells: they secretcytokines such as interferon gamma (IFNγ) and induce CMI and Th1 polarization. Recently, we
showed that CMI might be the principal component of the immune system in preventing infection
from E. tarda, intracellular bacteria found in fish. In this study, to elucidate the contribution of CTLs
in protection against E. tarda infection in fish, we used an adoptive transfer model established inclonal ginbuna crucian carp. We conducted a challenge test and analyzed CMI-related gene
expression. CD8α+ leukocytes were isolated from the kidney of E. tarda-sensitized (105 CFU/100 gbody weight [0.2 LD50]) ginbuna crucian carp on day 8 after injection using magnetic cell sortingwith anti-ginbuna CD8α monoclonal antibody. Donor cells (1.5 × 105 cells) were adoptivelytransferred to isogenic recipient fish via the caudal vein. The challenge test was performed with an
LD100 of E. tarda by i.p. injection one day after transfer: all fish receiving sensitized CD8α+ cells
survived, whereas some of the fish with non-sensitized CD8α+ cell transfer died and others showed
serious clinical signs. This result indicated that CTLs play an important role in protection against E.
tarda infection. IFNγ gene expression in the leukocytes of the head kidney, trunk kidney, spleen, andperipheral blood was analyzed in recipient fish on day 2 after challenge using quantitative PCR. The
expression levels of ifng1 and ifng2 in the leukocytes of all the tissues were higher in the fish
receiving sensitized CD8α+ cells than in the fish with non-sensitized CD8α+ cell transfer. Thus,
sensitized CTLs act as a major cell type for IFNγ-production in E. tarda-infected fish. Moreover,these finding suggested that sensitized CTLs induce CMI during the early stages of E. tarda
infection.

Three dimensional models of Signaling Lymphocyte Activation Molecules(SLAM), a receptor for morbillivirus in carnivores
K. Ohishi1,2 R. Suzuki3, T. Maeda1, E. Abe1, Y. Endo4, T. Nagamine5, H. Yamamoto6,
T. Yoshida1,2, T. Maruyama1,2
1Japan Agency for Marine-Earth Science and Technology, Japan, 2 School of Marine Biosciences,
Kitasato University, Japan, 3Protein Research Unit, National Institute of Agrobiological Science,
Japan, 4School of Veterinary Medicine, University of Kagoshima, Japan, 5Conservation & Animal
Welfare Trust Okinawa, Japan, 6Tsushima Wildlife Conservation Center, Japan

Morbillviruses belonging to the family Paramyxoviridae cause highly contagious diseases in humans
and animals. To date, six virus species including measles virus (MV) and canine distemper virus(CDV), have been identified in land and marine mammals. A notable biological feature ofmorbillivirus is its high level of host specificity. Signaling Lymphocyte Activation Molecule(SLAM), belonging to immunoglobulin superfamily, is the principal cellular receptor formorbilliviruses allowing their entry and propagation in the host animals, and is a major determinantof the host specificity. The immunoglobulin-like V (IgV) domain in the extracellular region of
SLAM, provides an interface to bind with morbillivirus H protein. The natural host for CDV is dogs,
but the host range has been recently expanded to include other wild carnivorous animals. CDV
caused more than 18,000 deaths of Baikal seals, and also induced fatal diseases to large felids suchas lion or jaguars. To compare the interface to the virus on SLAM in the carnivores, we havedetermined the nucleotide sequences of the IgV domains of SLAMs from two Japanese wildcat
species, Iriomote wildcat (Prionailurus bengalensis iriomotensis) and Tsushima leopard cat(Prionailurus bengalensis euptilurus), which are in danger of extinction. The sequences of the two
wildcats were completely identical in the domain. We generated the 3D models on the base of thehomology with proteins with known crystal structures: marmoset SLAM complexed with MV
hemagglutinin and human NK, T, B cell antigens. We found thirty-two amino acid residues on theinterface of SLAM IgV domain, which are potentially involved in the interaction with viruses. The
two species of Japanese wildcats possessed several specific residues (S67, K72, T76, S82, and A87)
among the thirty-two residues, and their interfaces showed a positively charged electrostatic surface
compared to other carnivores. This result may indicate that the Japanese wildcats are less sensitive
against CDV.

The Plasmablast-like Leukocyte in the Kidney of Fugu (Takifugu rubripes)
T. Odaka1,2, S. Tsutsui2, T. Miyadai1, R. Sugamata3, H. Suetake1,3, Y. Suzuki3,
O. Nakamura2
1

Faculty of Marine Bioscience, Fukui Prefectural University, Japan
2

School of Marine Biosciences, Kitasato University, Japan
3

Fisheries Laboratory, The University of Tokyo, Japan

The process of B cell differentiation into plasma cells, which are antibody secreting cells, has been
largely unexplored in fish. Research on the differentiation and regulation into the plasma cells
provides us with a better understanding of fish adaptive immunity. In this study, we isolated a unique
leukocyte population from fugu (Takifugu rubripes) kidney, which is the major immune organ inteleost. This leukocyte population showed phenotypic properties similar to those of mammalianplasmablasts. First, the adherent cells from the fugu kidney expressed IgM protein on their surface.
Flow cytometry showed that these cells were mainly composed of two populations: IgM+CD8α− cells
and IgM+CD8α+ cells. An reverse transcription PCR (RT-PCR) analysis demonstrated that the
purified IgM+CD8α− cells expressed secretory-type IgM as well as Bcl-6 and Blimp-1,
developmental marker genes for the B cell lineage, but not membrane-type IgM and Pax-5 genes. In
addition, we examined the cytokine expression profile of the IgM+CD8α− cells by RT-PCR. Most of
the cytokine genes were evidently not expressed by the IgM+CD8α− cells, whereas the IgM+CD8α+
cells expressed various cytokines. Western blotting confirmed that the IgM+CD8α− cells secreted
IgM protein. These results indicate that IgM+CD8α− cells are similar to the plasmablasts in
mammals. This is the first report isolating plasmablast-like leukocytes in fish species. Our data alsosuggest that the teleost kidney is an organ where B cells terminally differentiate into the plasma cells.

Session: Vertebrate Innate/Homeostatic Response
Expression of Type-I Interferon Gene Controlled by IRF3/7 Mediated through
TBK1 in Japanese FlounderJun-ichi Hikima1, Maki Ohtani1, Haruko Takeyama2, Tae-Sung Jung1, Takashi Aoki1, 3
1

Aquatic Biotechnology Center of WCU project, Gyeongsang National University,
Republic of Korea
2
Department of Life Science and Medical Bioscience, Waseda University, Japan
3
Consolidated Research Institute for Advanced Science and Medical Care,
Waseda University, Japan

Type I interferon (IFN-I) plays a very important role in the enhancement of antiviral innate
immunity. In a previous study, the IFN-I promoter region in Japanese flounder (Paralichthys
olivaceus), which initiates response to poly I:C, was identified and its transcriptional activity was
shown to be dramatically enhanced by IFN regulatory factor (IRF) 3. In this study, we found newfunctional differences between IRF3 and IRF7 in the IFN-I induction pathway. The IFN-I
transcriptional activity was significantly enhanced by IRF7 in HINAE (flounder natural embryo)cells when stimulated by poly I:C (a dsRNA mimic). To understand this function of IRF7,
TANK-binding kinase 1 (TBK1, Serine/threonine protein kinase) cDNA was cloned from Japaneseflounder. The deduced amino acid sequence of its domains was highly conserved with known
vertebrates and it was shown to be weakly induced in the whole kidney and HINAE cells after
stimulation by poly I:C and VHSV (viral hemorrhagic septicemia virus). In TBK1-overexpressing
HINAE cells, IFN-I transcriptional activity was sufficiently enhanced and the expression of
IFN-inducible gene (i.e. Mx and ISG15) was strongly induced. IFN-I activity was also induced bycotransfection with both TBK1 and IRF7, but not by cotransfection with IRF7 only. This evidence isvery similar to the IRF7 activation by poly I:C described above. To confirm the relationship between
IRFs and TBK1 in the IFN-I induction pathway, the transcriptional activities of IRFs were measured
after the cells were treated with the TBK1-inhibitor, BX-795. Activity of IRF7 mediated by TBK1 orpoly I:C was inhibited by BX-795 in a dose-dependent manner, but that of IRF3 was not.
Furthermore, IFN-I activity mediated through TBK1 was enhanced by Toll-like receptor 3 (TLR3)
cotransfection. These results suggested that the transcriptional regulation of Japanese flounder type I
IFN gene is regulated by IRF3 and IRF7, but its regulation by IRF7 is controlled by TBK1 after
TLR3 triggers the recognition of extrinsic dsRNA.

Poster Presentation

Session: Cytokine and chemokine
Earthworm extract possessed anti-inflammatory property through thedown-regulation of IL-8 transcription
Takashi Iwase, Shota Nomoto, Kazuo Komiyama
Department of Pathology, Nihon University, Japan
Earthworm is a member of Oligochaeta in the phylum Annelida. It was well known as a bite for themole, birds and others animals. Earthworm has been also recognized to use in oriental medicine.
Previous reports demonstrated that earthworm extract revealed to exhibit cytolytic activities,
proteolytic activities, anti-bacterial activities and anti-inflammatory property. However, precise
mechanism of these pharmaceutical effects was not fully clarified yet. We have examined the effectof earthworm (eisenia fetide) extracts on the cytokine expression in human epithelial cell lines andmouse macrophages. Human epithelial cells, HT-29 and Ca9-22 cells originate from the alimentarytract and mouse peritoneal macrophages were used in this study. The cells, stimulated by TNF andlipopolysaccharide, were co-cultured in various concentrations and times with earthworm extracts.
Cytokines, like IL-1, Il-6, IL-8 and TGF- were assessed by the RT-PCR, real time PCR andWestern blot analysis.
The extract clearly reduced IL-8 transcription level on the HT-29 and Ca9-22 cells following
TNF stimulation. While, other cytokine transcription did not significantly change. Western blot
analysis indicated that HT-29 and Ca9-22 cells decreased expression of the NF-kB, p38 and pErk1/2.
In addition, earthworm extract reveled similar effect to the mouse peritoneal macrophages, which
reduced the MIP-2 (functional IL-8 homologues in mouse) RNA transcription following stimulation
with LPS. Our results suggested that the earthworm extract have the anti-inflammatory activity due
to suppress the IL-8 secretion by MAPK inhibitory function.

Teleost IL-6 promotes antibody production through STAT3 signaling
via IL-6R and gp130
M. Kaneda1,2, T. Odaka1, H. Suetake1, T. Miyadai1
1

Faculty of Marine Biosciences, Fukui Prefectural University, Japan
2
Fukui Prefectural Fisheries Experiment, Japan (present address)

Antibodies play an important role in the adaptive immune system of teleost fish. When teleost fish
encounter pathogens, they increase secretion of specific antibodies into the blood and external
mucus. In mammals, antibody production is mainly regulated by interleukin-6, IL-6. Binding of IL-6
to the receptor complex consisting of IL-6 receptor (IL-6R) and glycoprotein-130 (gp130) stimulates
intracellular signal transduction, that is, phosphorylation of STAT3. STAT3 activated by IL-6 thendirectly activates IgM-gene transcription. Teleost IL-6 is upregulated after antigen stimulation;
therefore, we hypothesized that fish IL-6 contributes to antibody production during immune
responses against infections. In this study, the membrane and soluble forms of IL-6R homologues
cDNA were cloned in fugu (Takifugu rubripes). The cDNA of gp130 was also cloned. Three
STAT3-docking sites were found in the intracellular region of fugu gp130. Fugu IL-6R and gp130
were expressed in the membrane IgM+ (mIgM+) B cells of leukocyte populations in the kidney andperipheral blood, suggesting that fugu B cells are stimulated by IL-6. Recombinant fugu IL-6
(rfIL-6) increased the gene expression of secretary antibodies by mIgM+ B cells in vitro. Western
blotting analyses showed that the rfIL-6 activated STAT3 phosphorylation in B cells and a cultured
cell line cotransfected with fugu IL-6R and gp130. Multiple STAT-binding sites were found at the
fugu immunoglobulin heavy chain gene locus. These results indicate that fugu IL-6 enhances
antibody production in the B-cell lineage via gp130 and STAT3 signaling.

Session: Ig and other antigen receptors
Sequencing immunoglobulin variable regions of new antigen receptor genes inthe banded houndshark, Triakis scyllium
S. Suda1, I. Hirono1, H. Kondo1, T. Aoki2
1
2

Laboratory of Genome Science, Tokyo University of Marine Science and Technology, Japan
Consolidated Research Institute for Advanced Science and Medical Care, Waseda University,
Japan

Cartilaginous fish possess the immunoglobulin isotype new antigen receptor (IgNAR), which is aheavy chain homodimer and does not associate with the light chain. The IgNAR genes are encodedin the cluster configuration, each of which contains one variable (V) segment, some diversity (D)segments, one joining (J) segment and a constant domain. It was reported that IgNAR variable
regions have very high level of diversity in the complementarity determining region 3, where V, D
and J segments are rearranged. We have cloned the cDNA encoding IgNAR from the banded
houndshark (Triakis scyllium) and showed that this shark can produce a variety of IgNAR. Becauseof this, IgNAR can be a potential source of artificial single chain antibodies. In order to generate thesingle chain antibody efficiently, sequence information on the genes encoding IgNAR variable
regions are needed, we therefore cloned IgNAR variable region from the banded houndshark. The
genes encoding this region were amplified with specific primer sets designed using the cDNA
sequences. The fragments were amplified from genomic DNA extracted from the muscle andsubcloned into pGEM-T easy vector. We isolated and sequenced 27 clones. Twelve (12) clones wereabout 1.9kbp, and 2 clones were 1.4 kbp and the rest were 1.2 kbp long. Those showing the samelength were identical, and thus were grouped into 3 individual sequences. The V and J segments ofthese clones were almost identical, while the D segments of the genes were distinct. It was also
reported that the nurse shark (Ginglymostoma cirratum) and spiny dogfish (Squalus acanthias) haveseveral IgNAR genes. Therefore, these results suggest that the banded houndshark might have atleast 3 IgNAR genes. We are now evaluating the number of IgNAR genes by Southern blotting.

Session: Innate cellular immunity
Fatty acid binding protein 7 regulates the functions of murine macrophages
H. Miyazaki1, T. Sawada1, M. Harada1, M. Kiyohira1, N. Tokuda1,
Y. Adachi1, Y. Owada1
1

Department of Organ Anatomy, Yamaguchi University Graduate School of Mediciene

Fatty acid binding protein (FABP) is the family of cytoplasmic proteins with molecular weight of
14-15 kDa, having a phylogenically reserved structure. FABP serves as vehicle of water insoluble
fatty acids (FAs), and may consistently play important roles throughout animal evolution in theregulation of lipid metabolism, intracellular signaling mediated by FA metabolites, and gene
expression by nuclear receptors which require fatty acids as ligand. As it is known that dietary
polyunsaturated fatty acids affect the immune responses both in invertebrates and vertebrates,
members of fatty acid binding protein superfamily have been studied with relation to innate
immunity in both invertebrates and invertebrates. In mammal, FABP4 and 5 are generally
expressed in macrophages except for Kupffer cells (KCs) of liver which specifically express FABP7.
FABP4 and 5 have been shown to regulate the macrophage functions in adipose tissue and in theathelosclerotic plaque, but the role of FABP7 in KCs remained unknown. In this study, we
explored the functions of FABP7, a member of the FABP family that preferentially binds to omega-3
PUFA, in KCs. In the mouse liver injury model induced by CCl4 injection, FABP7 was detected in
the F4/80+ macrophages before and after the treatment. FABP7 deficient (KO) mice showed theincrease in the levels of serum ALT and AST, and infiltration of F4/80+ macrophages into damaged
area was significantly decreased compared to wild-type. The phagocytic activity of KCs against
intravenously injected apoptotic thymocytes (ATC) was significantly decreased in KO mice.
Interestingly, serum level of TNF-alpha was decreased in KO mice in D-GalN/LPS induced mouse
hepatitis models, and conversely, FABP7-overexpressing J774 cells showed increased activity ofanti-apoptotic cell phagocytosis and of LPS-induced TNF-alpha production. Taken together, it wassuggested that FABP7 is involved in the regulation of macrophage functions in mice.

Carp thrombopoietin in combination with kit ligand a induces the formation ofcolonies derived from unipotent progenitors for thrombocytes and bipotent
progenitors for thrombocytes and erythrocytes
Y. Sugie1, F. Katakura1, T. Yamaguchi1, T. Kato2, T. Yabu1, T. Moritomo1
T. Nakanishi1
1
2

Laboratory of Fish Pathology, Department of Veterinary Medicine, Nihon University, JAPAN
Laboratory of Molecular Physiology, Department of School of Education, Waseda University,
JAPANIn mammals, thrombopoietin (TPO) and Kit ligand (KitL; stem cell factor) are primary regulators ofthrombopoiesis. TPO stimulates megakaryocyte progenitor cells (CFU-MK) and induces
megakaryocyte colony formation in vitro in a semi-solid medium. Megakaryocyte colony formationis enhanced by TPO in combination with KitL, but not by KitL alone. To understand thenon-mammalian thrombopoiesis, we produced recombinant carp TPO and kit ligand a (kitla), anddeveloped a culture system capable of examining quantitatively and qualitatively analyzing ofthrombocyte progenitors in carp. About 100,000 hematopoietic cells from carp kidney were cultured
in a semi-solid methylcellulose medium. TPO induced colony formation in a dose-dependent
manner. The maximum number of colonies was 110 - 150 per plate and all colonies were small (type1), which were 50 to 100 μm in diameter. However, kitla alone did not induce colony formation. In
addition, in the presence of both TPO and kitla, 300 - 380 colonies formed, the majority of whichwere type 1 and several larger colonies (200-500μm in diameter) also formed. The larger colonies
were divided into two types (types 2 and 3) based on their morphologies. Using RT-PCR, Type 1
cells were found to express a thrombocyte marker c-mpl (TPO receptor), type 2 cells were found to
express both c-mpl and another thrombocyte marker cd41 (a cell surface glycoprotein ofthrombocyte lineage cells) and type 3 cells were found to express both of these markers and threeerythrocyte markers ( β -globin, epo-r and gata1). From these gene expression patterns, weconsidered that type 1 and type 2 colonies were derived from unipotent progenitors forthrombocytes. Type 2 colonies were derived from more immature progenitor cells than those of type
1 colonies. In contrast, type 3 colonies were derived from bipotent progenitors for thrombocytes anderythrocytes. In summary, TPO induced thrombocyte colony formation in a dose-dependent manner.
The combination of TPO and kitla resulted in a synergistic enhancement of number of thrombocytecolonies, and induced the formation of mixed-type colonies derived from bipotent progenitors forthrombocytes and erythrocytes.

Session: Invertebrate hematocyte
Morphological Diversity of Crustacean Hemocytes
M. Kondo, S. Yasumoto, Y. Takahashi
Department of Applied Aquabiology, National Fisheries University, Japan
Morphological characteristics of crustacean hemocytes were examined by light microscopy afterstaining with May-Grünwald stain using acid phosphate buffer (1/15 M, pH5.0) as a diluent. Only asingle type of hemocyte (granulocyte) was observed in the primitive crustaceans like branchiopods(brine shrimp Artemia salina (A), fairy shrimp Branchinellites kugenumaensis (B), tadpole shrimp
Triops numidicus (C), clam shrimp Leptestheria kawauchieusis (D), water flea Daphnia pulex (E))
and maxillopods (fish louse Argulus japonicus (F), seed shrimp Vargula hilgendorfii (G), ship
barnacle Lepas anatifera (H), balanomorph barnacle Megabalanus rosa (I)). Staining characteristics
of granules were different among these species. Two types of granules were observed in thehemocytes of A and G. The former had eosinophilic granules (EG) and chromophobic granules(CG), and the latter had basophilic granules (BG) and CG. In other species, the hemocyte had one
type of granules (CG, excluding D (amphophilic granules)). The primitive malacostracan species,
leaf shrimp Nebalia japonensis had only one type of hemocyte containing two types of granules (EG,
BG). We call these species as monohemocytic crustaceans. On the other hand, plural hemocyte types
were detected in the advanced malacostracans such as stomatopods (Oratosquilla oratoria,
Harpiosquilla harpax), isopods (Ligia exotica, Armadillidium vulgare, Asellus hilgendorfii),
amphipod (Melita koreana), and decapods. These species (polyhemocytic crustaceans) wereclassified into five groups (1-5) as below: 1, eight hemocyte types group (basophilic fine granularcell, basophilic plasma cell, basophilic granulocyte, chromophobic small granulocyte, chromophobic
large granulocyte, basophilic and eosinophilic granulocyte, eosinophilic granulocyte (EGC; type 1
and 2)) including stomatopods, isopods (L. exotica, A. vulgare), amphipod, and decapods (Penaeidea,
Thalassinidea, Brachyura); 2, four hemocyte types group (small basophilic cell, chromophobic
granulocyte (CGC), eosinophilic small granulocyte (ESG), eosinophilic large granulocyte (ELG))
including decapods (Astacidea, Palinuridea, Anomura); 3, three hemocyte types group (CGC, ESG,
ELG) including decapods (Astacidea, Brachyura); 4, two hemocyte types group (CGC, EGC)
including isopod (A. hilgendorfii) and decapods (Stenopodidea, Caridea); 5, two hemocyte types
group (ESG, ELG) including decapod (Brachyura). Based on these results, we make a speculation
about the evolutional process of crustacean hemocytes.

Comparative morphology of hemocytes in deep-sea symbiotic bivalves, three
Bathymodiolus mussels and two Calyptogena clams
A. Tame1, T. Yoshida2, K. Ohishi2, T. Maruyama2
1

2

Marine Works Japan Ltd., Japan
Japan Agency for Marine-Earth Science and Technology, Japan

Hemocytes are involved in the defense system against microbial infections. Deep-sea bivalves areoften dominant in deep-sea chemosynthetic ecosystems and mostly harbor methane-oxidizing and/or
sulfur-oxidizing bacteria in their epithelial cells of gill tissues. Defense system probably plays
important roles in the symbiotic interaction between the bacteria and the host animals. However,
little is known about the defense system of deep-sea symbiotic bivalves. To understand the defense
system in deep-sea symbiotic bivalves, we examined the hemocytes, which are thought to play a keyrole in the defense system, in five deep-sea symbiotic bivalves, Bathymodiolus japonicus, B.
platifrons, B. septemdierum, Calyptogena okutanii and C. phaseoliformis, and classified them onmorphological criteria. To see the functional difference among hemocytes, a phagocytosis assay
using a FITC-labeled Escherichia coli was performed. Hemogram parameters (cells concentration,
number and proportions of hemocyte types) and lectin-binding using four lectins (WGA, Con A,
PHA-L, SBA) were also studied and compared in the bivalves. The results indicated that all of thebivalves have three types of hemocytes, a hemocyte type without granule and two types with
granules. The two types were granular cell and hyaline cell. The hemocyte without granules wascalled agranular cell in Bathymodiolus mussels, and was smaller than the granular cell and hyalinecell. In two Calyptogena clams, this type of hemocyte was the red-blood cell having hemoglobin. In
any of the examined bivalves, the granular cell showed the highest phagocytic activity and theresultant food vacuoles fused with lysosomes. The hyaline cell also showed the phagocytic activity
with less extent but showed no fusion with the lysosome. The agranular cell and the red-blood cellshowed no phagocytic activity. These results suggested that two types of granulocyte have thephagocytic activity which is the fundamental activity in the defense system against microbial
infection in deep-sea symbiotic bivalves. The function of the agranular cell is still to be studied but
the red-blood cell has hemoglobin, the oxygen carrier. Morphological study similarity suggested thatthe red-blood cell in Calyptogena clams is originated from the agranular cell.

Session: Invertebrate immune response
Slow transport of foreign objects to the distal end of the sea urchin larval arms
Taku Hibino, Aki Kanamori, Daisuke Naka
Faculty of Education, Saitama University, JapanThe sea urchin and other echinoderm larvae have large advantages for studying developmental andcomparative immunology, because the larva has a simple and transparent body consisting of small
number of cells. The monumental findings using a kind of this larva are achieved by Iliya
Metchnikoff. He found phagocytosis from the starfish larva when he injected a rose throne, a spineof a sea urchin and a glass tube. He also reported that the phagocytosis reaction consists merely in an
accumulation of mesodermic phagocytes around the foreign body. However, he was not shown what
happens after phagocytosing these undigested objects. If bacteria E. coli transformed by GFP-vector
are injected into the blastocoel of a pluteus larva, GFP signal in the phagocytosed bacteria quickly
get weak, showing the digestion of phagocytosed bacteria. In the case of undigested objects, adifferent defense system from digestion is expected to work, but it is still unveiled. In the present
study, to find the next step of phagocytosis, we injected fluorescent latex beads and other objects
with a micro-needle into the blastocoel of the sea urchin pluteus larva, and then tracked the beads
during 5 days after injection. The beads were dispersed in the blastocoel just after injection, whose
fluorescent signals were gradually reduced in the blastocoel. Instead, some of the beads were aligned
on trunck spicules and arm spicules, which were prominent in the larvae 4 and 5 days after injection.
The beads along with the spicules were migrated to the distal end of the arms revealed by an hourobservation for each larva (n=10) under the microscope. Next, we injected FITC-labeled zymosanand RITC-labeled E. coli in the blastocoel of the larvae. A part of them are slowly transported to the
distal end of the arms as well. It is still unknown where the foreign objects at the distal end go. We
propose that the spicules of the sea urchin larva are functional for distancing foreign objects from thetrunk containing a digestive organ and an adult rudiment to the distal end of the arms.

Characterization of a downstream component of Drosophila receptor guanylate
cyclase activating innate immune response
Hirotaka Kanoh, Takayuki Kuraishi, Hiroki Ishikawa, Shoichiro Kurata
Graduate School of Pharmaceutical Sciences, Tohoku University, Japan
Drosophila receptor-type guanylate cyclases (rGCs) are novel immune related receptors identified
from a genome-wide gain-of-function screening. rGCs are conserved in human as natriuretic peptide
receptors producing cGMP in response to intrinsic ligands. Overexpression of rGCs induces
Drosomycin, an antimicrobial peptide, through cGMP and downstream components of the Toll
receptor, such as dMyD88, in a Toll receptor-independent manner in Drosophila. These results imply
the existence of novel cGMP-mediated signaling pathway in immune activation. To elucidate its
signaling mechanisms, we identifed a cGMP-dependent signaling molecule which acts downstreamof rGCs in Drosophila larvae and characterized its function in Drosophila culture cells. Our results
showed that the induction of RNAi targeting a cGMP-dependent protein kinase (cGK) completely
abrogated rGC-induced Drosomycin expression in larvae. Overexpression of cGK and rGC strongly
augmented the Toll-pathway-dependent Drosomycin expression in both larvae and Drosophila
culture cells. Site-direct mutagenesis analysis showed that kinase-dead mutants of cGK failed toactivate Toll-pathway. Furthermore, flies inducing RNAi targeting cGK were susceptible to infectionof Gram-positive bacteria. These results suggest that Drosophila cGK is required for host defense
and mediates the cGMP-dependent immune activation downstream of Drosophila rGCs.

Session: Pattern Recognition Molecules
Evolutional Conservation of Primordial Functions and Transcriptional Controlof LGP2 Gene in Japanese Flounder, Paralichthys olivaceus
M. Ohtani1, J. Hikima1, M. K. Yi1, T.S. Jung1, H. Takeyama2, T. Aoki1, 3
1

Aquatic Biotechnology Center of WCU project, Gyeongsang National University, South Korea
2
Department of Life Science and Medical Bioscience, Waseda University, Japan
3
Consolidated Research Institute for Advanced Science and Medical Care,
Waseda University, Japan
LGP2 (laboratory of genetics and physiology 2), one of the pattern-recognition receptors, plays avery important role in antiviral response to induce production of type I interferon (IFN) through therecognition of cytosolic viral RNAs in the innate immune system. Although the expression of LGP2mRNA in mammals and teleosts are strongly induced after virus infection or poly I:C stimulation,mechanism of transcriptional control of LGP2 gene is still unknown. Currently, LGP2 gene inJapanese flounder (Paralichthys olivaceus) was cloned. Japanese flounder LGP2 gene spanned 5,474
bp containing 12 exons and 11 introns. The expression of LGP2 mRNA dramatically increased inwhole kidney of Japanese flounder infected with VHSV (viral hemorrhagic septicemia virus) 3 days
post-infection and in whole kidney leukocytes stimulated with artificial double-stranded RNA (poly
I:C) in vitro. Japanese flounder LGP2 has strong antiviral activities against VHSV, HIRRV (hirame
rhabdovirus) or IPNV (infectious pancreatic necrosis virus) infected flounder natural embryo(HINAE) cell line. However when the RD (regulatory domain) of the LGP2 was deleted this function
was lost. In addition, the expression of Mx and ISG15 in LGP2-overexpressed HINAE cells werestrongly induced by poly I:C co-transfection but not by poly I:C-addition into the culture medium.
The transcriptional control of Japanese flounder LGP2 gene was identified and its transcriptional
activity was also analyzed by luciferase reporter assay. Numerous canonical motifs of
IFN-regulatory factors (IRFs) were found in the 5’-upstream region (-1,337 bp) of LGP2 gene.
Reporter assay showed that the poly I:C-responsive region regulating LGP2 transcription was
identified at the location of -506 to -398. The transcriptional activity of poly I:C-responsive region
was strongly enhanced by IRF3, which could bind to IRF3#3 motif located at -480, suggesting that
LGP2 transcriptional control is probably involved in IRF3 function. Furthermore, LGP2 promoter
was enhanced by VHSV infection in HINAE cells using GFP expression construct regulated by the
LGP2 promoter including the poly I:C-responsive region. These results suggested that Japaneseflounder LGP2 acts as a cytosolic viral RNA sensor, and the functions are conserved with those of
mammalian LGP2 in the induction of antiviral response in the innate immune system.

Session: T-cell, MHC, and Ag-presentation
Different Evolutionary Patterns of Three Alpha Domains of
the MHC Class IA Genes
M. I. Nonaka, M. Nonaka
Department of Biological Sciences, The University of Tokyo, JapanThe major histocompatibility complex (MHC) genomic region contains many genes encoding
various proteins, which play pivotal roles in the immune defense system of jawed vertebrates. MHC
class I alpha chain gene (IA gene) is one of these and its product composes a class I molecule withbeta-2 microgrobulin (B2M). MHC class I molecules function in the presentation of intracellularpeptide antigens to cytotoxic CD8+ T cells that kill those cells directly. Most vertebrates possess
multiple IA loci and most of them are highly polymorphic possibly to be able to adapt to various
kinds of antigens. Because genetic events such as gene duplication, deletion, recombination, and/or
gene conversion have occurred frequently in these genes, the analysis of their evolutionary
relationships between distantly related species has difficulties. We performed phylogenetic analyses
using the nucleotide sequences of three alpha domains of the classical and non-classical IA genes
independently, and found that these three domains have quite different evolutionary histories. In theanalysis of the teleost fishes, the 1 domains generated four deeply diverged locus-specific lineages
with high bootstrap values. Two of them contained the salmonid and medaka IA genes, unveiling theorthologous relationships between IA loci of salmonids and medaka, which diverged approximately
260 million years ago. In contrast, the 3 domains clustered by species or fish groups, regardless ofclassical or non-classical gene types, suggesting that this domain was homogenized in each species
during prolonged evolution. On the other hand, the 2 domains formed no apparent clusters with the1 lineages or with species, suggesting that they were diversified partly by interlocus geneconversion and that the 1 and 2 domains evolved differently. Although the primate IA genes
evolved locus-specifically in all three domains, the homogenization of the 3 domain amongclassical and non-classical IA genes is also seen in some other vertebrates. Such distinct evolutionary
patterns of three alpha domains might have contributed to the diversification of the peptide-bindingsite composed of the 1 and 2 domains, retaining the potential for CD8 binding as well as B2M
binding in the 3 domain, during long-term evolution.

Characterization and expression analysis of CD4 and CD8 genes in yellowtail,
Seriola quinqueradiata
K. Araki1, Y. Shimono1, M. Yamasaki2, T. Matsuyama3, S. Yanagi3, T. Murase3
A. Yamamoto1
1

2Faculty of Fisheries, Kagoshima University, JapanThe United Graduate School of Agricultural Sciences, Kagoshima University, Japan
3
National Research Institute of Aquaculture, Fisheries Research Agency, Japan
4
Kagoshima Prefectural Fisheries Technology and Development Center, Japan

CD4 is a membrane glycoprotein that is mainly expressed on helper T (Th) cells and acts as a TCR
coreceptor by binding to MHC class II molecules on antigen-presenting cells in mammals. Fishpossess 2 types of CD4-like molecules, which are CD4-1 and CD4-2. The fish CD4-1 protein as wellas the mammalian CD4 protein consists of 4 extracellular immunoglobulin (Ig)-like domains, atransmembrane domain and a cytoplasmic domain, whereas the fish CD4-2 proteins has only 2 or 3
Ig-like domains and a membrane proximal connecting peptide in the extracellular region.
Mammalian CD8 is usually expressed on the surface of cytotoxic T lymphocytes (CTLs) as aheterodimer of α and β chains. In this study, we have cloned CD4-1 and CD8α homologues from thethymus of the yellowtail (Seriola quinqueradiata) by performing RACE. The yellowtail CD4-1
cDNA is 2122 bp long and encodes 467 amino acids. The deduced amino acid sequence of yellowtail
CD4-1 had 4 extracellular Ig-like domains, a transmembrane domain, and a p56lck-binding motif inthe cytoplasmic domain, similar to the CD4-1 of other fish species and mammalian CD4. The
yellowtail CD8α cDNA is 1649 bp long and encodes 217 amino acids. The deduced amino acidsequence of yellowtail CD8α cDNA was structurally similar to those of other vertebrates.
Phylogenetic analysis showed that yellowtail CD4-1 and CD8α were clustered with other vertebrate
CD4/CD4-1 and CD8α, respectively. Tissue-distribution analysis showed that yellowtail CD4-1 andCD8α as well as TCRα were predominantly expressed in lymphoid tissue, i.e., in the thymus, headkidney, and trunk kidney, and in mucosal tissue, i.e., the gill and intestine. In addition, the expressionof these genes was detected in surface Ig-positive leukocytes but not in surface Ig-negative
leukocytes. Moreover their expression levels were upregulated in the kidney of fish infected by
Mycobacterium sp. at 2 months after infection. This result suggests that Th cells and CTLs play arole in protection against mycobacterial infection in the yellowtail.

Session: Vertebrate adaptive immune responce
Characterization of culture supernatant of Streptococcus parauberis: hemolytic
factor and leucocyte-stimulation in Japanese flounder
Haruka Tsukamoto1, Yutaka Fukuda2, Tomonori Somamoto1, Miki Nakao1
1

2

Department of Bioscience and Biotechnology, Kyushu University, Japan
Fisheries Research Division, Oita Prefectural Agriculture, Forestry and Fisheries ResearchCenter, Japan

Two serotypes of Streptococcus parauberis, pathogens of streptococcosis in Japanese flounder(Paralichthys olivaceus), have caused severe losses of the livestock, due to the lack of effective
vaccine and its resistance to antibiotics approved for flounder culture. These strains show different
virulence, but their mechanisms of their pathogenicity are totally unknown, and identification ofvirulence factors are needed for effective vaccine development. The present study, therefore, wasaimed at clarifying effect of S. parauberis culture supernatant on flounder. Since the diseased
flounder shows anemia, we examined the effects of S. parauberis on flounder peripheral blood. As aresult, S. parauberis -injected flounder showed decreased number of peripheral erythrocytes,
suggesting that S. parauberis may produce a hemolytic factor. To characterize the hemolytic factor,
a supernatant of S. parauberis culture (25˚C for 48h in Todd Hewitt broth) was mixed with sheep
erythrocytes, resulting in significant level of hemolysis. The hemolytic factor was stable on heating
at 100˚C for 10 min, and passed through an ultrafiltration membrane with a 5 kDa cut off limit(Amicon Ultra15), indicating that the factor is not proteinous hemolysin but a heat-resistant low
molecular mass substance. We also investigated the effects of the culture supernatant on flounderleukocytes. Peripheral leukocytes were separated from flounder and carp using Percoll discontinuous
density gradient centrifugation and cultured with the supernatant in 96-well plate. The stimulated
flounder cells showed agglutination and significant proliferation as assayed by BrdU uptake, while
carp cells did not. These results suggest that S. parauberis secrets a mitogen specific for flounderleucocytes. Characterization of the mitogen and resulting leucocyte response is in progress.

Session: Verterbate Innate/Homeostatic Response
Candidate key molecule(s) determining host specificity of parasite on fugu,
Takifugu rubripes
S. Tasumi1, A. Yamaguchi2, Y. Hirabayashi1, S. Kido1, K. Kobayashi2, W. Kai1,
S. Hosoya1, S. Tsutsui2, O. Nakamura2, H. Suetake1, K. Kikuchi1, Y. Suzuki1
1

2

Fisheries Laboratory, The University of Tokyo, Japan
School of Marine Biosciences, Kitasato University, JapanFugu, Takifugu rubripes, is susceptible to infection with monogenean parasite, Heterobothrium
okamotoi, which parasitize gills. Intriguingly, relative species of fugu, Takifugu niphobles, is not
susceptible to the parasite although genetic distance between T. rubripes and T. niphobles is so closethat their F2 hybrids are viable. Genetic methods are applicable to T. rubripes for studying many
biological aspects including host-parasite interactions since its genomic sequences have been
analyzed and its comprehensive genetic map is also available. In the present study, we used F2
individuals for genome-wide quantitative trait loci (QTL) analysis for parasite burden 21 days after
challenge. As the result, we found a single major QTL peak on linkage group 9, and the region of the
95% confidential intervals contained 268 annotated genes. As this number of genes is still large, wehave been conducting two additional experiments, i.e., cell surface display and subtractive PCR. By
cell surface display, we successfully expressed two selected model membrane proteins on the surface
of insect cells (High Five). By using this platform, we are now screening candidates for receptors
expressed in gills, which specifically bind to ligands on H. okamotoi. By subtractive PCR, we aretrying to find genes, whose expression levels in gills are different between T. rubripes and T.
niphobles. So far we found numbers of candidate genes including pufflectin. Pufflectin is a
D-mannose specific lectin originally found in the skin mucus of fugu, and is expressed in skin as
well as in gills. By semi-quantitative PCR and western blotting, we found that the expression of
pufflectin is certainly much higher in T. rubripes than in T. niphobles. In our previous report, we
showed that pufflectin could bind to body surface of H. okamotoi. These data suggests that pufflectin
is one of the strong candidates for important factors determining host specificity of H. okamotoi. We
are now conducting experiments to obtain direct evidences showing the relationship between theexpression level of pufflectin and host specificity of H. okamotoi.

ISDCI 2012 Program Timetable
Date
July
9 Mon 11:00
13:50 - 14:00
14:00 - 15:00
15:00 - 15:30
15:00 - 17:50

10 Tue18:30 - 20:00
20:00 - 22:30
9:00 - 10:00
10:00 - 10:30
10:30 - 11:30
11:30 - 12:30
12:30 - 13:30
13:30 - 15:30

15:30 - 16:00
16:00 - 17:00

11 WedBreak
Room <Navis - C>
Cytokines and Chemokines - 1

15:30 - 18:00

Welcome Reception <Tinga-Tinga>
DCI Editorial Board meeting < Kusu >Plenary Lecture by Dr. Zeev Pancer < Argos D >BreakRoom <Navis - C>
Room <Navis - B>
Cytokines and Chemokines - 2 10:30 - 12:30 Pattern Recognition
Antiviral Immunity - 1
Molecules - 1
Lunch < Argos E-F >Pattern Recognition
Antiviral Immunity - 2
13:30-14:30
Molecules - 2
14:30 - 15:30 Lectins - 1
Break
Antiviral Immunity - 3
16:00 - 16:40 Lectins - 2
16:40 - 17:20 Antimicrobial
peptides/proteins - 1

8:45 - 9:45
9:45 - 10:15

Plenary Lecture by Dr. Claudia Kemper < Argos D >BreakRoom <Navis - C>
Vaccine and Adjuvant

11:15 - 11:55

Room <Navis - B>
Antimicrobial
peptides/proteins - 2
Innate Cellular Immunity - 1

orFree Afternoon

10:15 - 11:15

12:15 - 13:15

Lunch < Argos E-F >13:15 - 17:30
18:15 - 21:40

Excursion (Optional)
Dinner Cruise (Optional)

9:00 - 10:00
10:00 - 10:30

Plenary Lecture by Prof. Arturo Zychlinsky < Argos D >10:30 - 12:30

Vertebrate Adaptive Immune
Response - 1

12:30 - 13:30
13:30 - 14:30

Lunch < Argos E-F >Vertebrate Adaptive Immune
Response - 2
Vertebrate Innate andHomeostatic Response - 1
BreakVertebrate Innate andHomeostatic Response - 2

15:10 - 15:40
15:40 - 17:40

Break
Room < Navis - C >10:30 - 12:30

Room < Navis - B >Innate Cellular Immunity - 2

13:30 - 15:10
Invertebrate Hemocytes -1

15:40 - 16:00
16:00 - 17:00

Invertebrate Hemocytes -2
Invertebrate Immune
Response - 1

18:00 - 20:00

ISDCI Exec. Meeting

9:00 - 10:00
10:00 - 10:30

Plenary Lecture by Prof. Thomas Boehm < Argos D >BreakRoom <Navis - C>
10:30 -11:10 Vertebrate Innate and10:30 -12:30
Homeostatic Response - 3
11:10 - 12:30 T cell, MHC, and antigenpresentation -1
12:30 - 13: 30 Lunch < Argos E-F >13:30 - 14:50 T cell, MHC, and antigen13:30 - 14:50
presentation -2
14:50 - 15:20 Break
15:20 - 16:40 T cell, MHC, and antigen15:20 - 16:20
presentation -3
17:00 - 18:00 ISDCI General Meeting < Navis -BC >19:30 - 22:30

Banquet < Argos E-F >13:00 - 13:50

JADCI Business Meeting

10:30 - 12:30

Room <Navis - A>
Ig and other antigenreceptors-1

Room <Navis - B>
Special Symposium"Recognition Mechanisms –Current Topics in
Comparative Biology –"Poster Session <Argos C >14:30 - 15:10

13 Fri

Registration Starts
Welcome Address
Plenary Lecture by Prof. Shoichiro Kurata < Argos D >17:30 - 20:30

10:15 - 12:15

12 Thu(updated 2012 June 16)

Room <Navis - B>
Invertebrate Immune
Response - 2

Invertebrate Immune
Response - 3
Invertebrate Immune
Response - 4

13:30-15:30

16:00 - 17:00

Ig and other antigenreceptors-1

Complement andComplement-like factors - 1

Room <Navis - A>
10:15 - 12:15

Complement andComplement-like factors - 2

Session allocation of abstracts
Updated 2012 June 19
Session Name and Date
Special Symposium"Recognition Mechanisms – Current Topics in

Day
Mon

* Each oral presentation will have 20 min (15 min talk & 5 min discussion).
* All the posters presentation will be held on Tuesday 10th.
* Presentations by JADCI members are indicated by ★
Time
ID No.
Presenter
Title
15:30 SS-1
B-L. Lee
Recognition of Pathogenic or Symbiotic Bacteria and Immune Responses in Insects
15:55 SS-2
R. Furukawa
Immunological Recognition in Starfish Larvae and Adults: The Alteration of Immune Cells Via Metamorphosis
16:20 SS-3
D. Kojima
OPN5, a Photosensory Protein for Mammalian Ultraviolet Photoreception
Ligand recognition and signal transduction of GPCRs and Toll-like receptors of the protochordate, Ciona intestinalis: what is
16:45 SS-4
H. Satakeevolutionarily common or unique?
17:10 SS-5
M. Yoshikuni
Comprehensive Analysis of Neuropeptides of Marine Invertebrates
17:35 SS-6
T. Sakurai
Specificity and Sensitivity of Sex Pheromone Perception in the Silkmoth Bombyx moriSession Name and Date
Antimicrobial peptides/proteins (AMP)

Day
Tue

Time

ID No.
16:40 AMP-O1
17:00 AMP-O2
10:15 AMP-O3
10:35 AMP-O4
10:55 AMP-O5
AMP-P1
AMP-P2
AMP-P3
Poster
AMP-P4
AMP-P5
AMP-P6

Presenter
Xinhua ChenJames D. Pask
Hai-Peng Liu
K. Wang
C. Li
K. ShenM. GuoS. Wei
S. Hipolito
Bo-Hye Nam
C.H. Bigger

Title
Analysis on the gene diversity of hepcidin gene in large yellow croaker
Normal constitutive secretion of skin peptides defends northern leopard frogs (Rana pipiens) against chytridiomycosis
Antibacterial responses in the crab Scylla paramamosain
Scygonadin expression in Scylla paramamosain potentially associated with reproductive immunity
Two novel families of antimicrobial peptides in the green sea urchin Strongylocentrotus droebachiensis
Escherichia coli DNA enhances the expression of attacin in Drosophila hemocyte cell line Mbn-2
Antiviral effects of beta-defensin derived from orange-spotted grouper, Epinephelus coioides
Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides
Role of Marsupenaeus japonicus crustin-like peptide against Vibrio penaeicida and white spot virus infection
An antimicrobial histone H1-like protein and its gene from the testes of Paralichthys olivaceus
Antimicrobial activity in aqueous extracts of the caribbean octocoral Swiftia exserta.

Session Name and Date
Antiviral immunity (AVI)

Day
Tue

Time

Presenter
Chu-Fang Lo
Apiruck
Watthanasurorot
Mei-An Su
S. Mangkalanan
J-X Wang
T. Somamoto
P. H. Pham
Fang-Huar Ngou
S. PatelJoeri Kint
B. Gorgoglione
C. Langevin
C.-Y. Chang
Y. J. Lee
T.Y. Chen
T.Y. Chen
H. Huang

Title
Spawning stress triggers WSSV replication in brooders via the activation of shrimp STATID No.
11:30 AVI-O1
11:50 AVI-O2
12:10 AVI-O3
13:30 AVI-O4
13:50 AVI-O5
14:10 AVI-O6
14:30 AVI-O7
14:50 AVI-O8
15:10 AVI-O9
16:00 AVI-O10
16:20 AVI-O11
16:40 AVI-O12
AVI-P1
AVI-P2
AVI-P3
AVI-P4
AVI-P5

Poster

AVI-P6

Rong-yuan Chen

AVI-P7

Shau-Chi Chi

Session Name and Date

Day

Time

★
★
★

A gC1qR Prevents White Spot Syndrome Virus Replication in the Freshwater Crayfish Pacifastacus leniusculus
The Litopenaeus vannamei LvRheb Protein is Involved in White Spot Syndrome Virus (WSSV) Pathogenesis
Hemocytic Response in Persistent WSSV Infection in the Mud Crab, Scylla olivacea
Prohibitin interacts with VP28 and prevents WSSV replication in red swamp crayfish, Procambarus clarkii
Functions of CD8-positive and CD4-positive lymphocytes against virus-infection in ginbuna crucain carp
Viral hemorrhagic septicemia virus genotype IV is differentially virulent toward Rainbow trout gills and monocyte/macrophage
Virus-Host Interactions of Nervous Necrosis Virus in Grouper GK cell by Transcriptomic Analysis
Nodavirus infection induces up-regulation of IFNgamma and T-helper cell markers.
Mechanism of suppression of type-I interferon signaling by the chicken infectious bronchitis coronavirus
ASSESSMENT OF THE IMMUNE RESPONSE MODULATION BY CO-INFECTIONS: PKD/VHS EFFECTS IN BROWN TROUT
Restriction mechanisms of fish rhabdoviruses by the zebrafish ISG15 protein
Differential expression profiles of orange-spotted grouper under grouper iridovirus infection
Identification of the Envelope Protein VP51B of Shrimp White Spot Syndrome Virus (WSSV)Grouper HSP90 Interacts with Nodavirus Coat Proteins and Participates in Nodavirus Replication
Identification of Leucine Zipper Domain of Antiviral Mx Protein Interfere with Nodavirus Coat Protein Localization
The study of protein-protein interaction between white spot syndrome virus and glucose transporter Glut1
Identification of genes differentially expressed in in vitro haematopoietic tissue stem cells of Cherax quadricarinatus in response to
WSSV infection
Betanodavirus-induced IFN response and Mx expression makes barramundi (Lates calcarifer) more resistant to red sea bream iridovirus
infection

AVI-P9Apiruck
Watthanasurorot
Youhua Huang

MAPK signaling pathway was involved in Singapore grouper iridovirus (SGIV) infection

ID No.

Presenter

Title

AVI-P8★
★
★

New insights into extracellular function of TCTP as an antiviral infection in Pacifastacus leniusculus

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Complement and Complemet-like factors
(CMP)

Tue

Wed

Session Name and Date
Cytokine and chemokine (CCK)

Day
Mon

Tue

Session Name and DateIg and other antigen receptors (IGR)

Day
Tue

16:00 CMP-O1

Irene Söderhäll

TEPs (thioester containing proteins) and ficolins; structure and function in a crustacean

16:20 CMP-O2
16:40 CMP-O3
10:15 CMP-O4
10:35 CMP-O5
10:55 CMP-O6
11:15 CMP-O7
11:35 CMP-O8
11:55 CMP-O9
CMP-P1
Poster
CMP-P2

A. Tassanakajon
T. Sasaki
T.Fujita
Vo Kha Tam
Jun LiT. Yamaguchi
P. Kopáček
K. Tagawa
Rui LiM. G. Castillo

A Vital Role of Alpha-2-Macroglobulin in Prevention of Bacterial-Mediated Fibrinolysis in the Blood Coagulation System of Shrimp
Cytolytic factor in the mosquito
MBL-associated serine protease (MASP)-1/3 promotes the activation of the alternative complement pathway
Isotypic Diversity in the Ontogenetic Expression of the Complement Component in the Common Carp (Cyprinus carpio)
Survivability of Edwardsiella tarda in Fish Serum Relates to Bacterial Surface LPS
Identification of Pattern-Recognition Molecule in Hagfish Complement System
Functional Genomics of Tick Thioester-Containing Proteins and Other Complement-Related Molecules
Microbe-specific C3b deposition in the horseshoe crab complement system in a C2/factor B-dependent or -independent manner
Unique Functions of MASPs in Amphioxus
Identification and Initial Characterization of a Thioester-Containing Protein (TEP) in the Squid Euprymna scolopes

Time

ID No.
15:30 CCK-O1
15:50 CCK-O2

Presenter
D. Malagoli
John Han-You Lin

16:10 CCK-O3
16:30 CCK-O4
16:50 CCK-O5

Suhee Hong
H. Suetake
H. Korenaga

Title
Drosophila Helical factor acts as an inducible cytokine in S2 cells
The bioactivity of teleost IL-6: orange-spotted grouper (Epinephelus coioides) IL-6 induce Th2 cell differentiation pathway and antibody
production
Two types of TNFa in fish: cloning, expression and functional characterization of the third TNFa in rainbow trout
A chemokine expressed in the secondary lymphoid organs of fugu (Takifugu rubripes)
Identification, characterization and expression analysis of IL-17 receptors, Act1 and TRAF6 genes in Japanese pufferfish (Takifugu
rubripes)
The two interferons of Large yellow croaker
MOLECULAR AND FUNCTIONAL CHARACTERIZATION OF IL12 FAMILY MEMBERS IN SALMONIDS
Phylogenetic and functional diversity of CXC chemokines and their receptors in cyprinids
Rainbow trout CK1 signals through CCR6 in B lymphocytes and is involved in the recruitment of IgT+ cells to mucosal sites
Identification of a novel CCR7 gene in rainbow trout with differential regulation in the context of mucosal or systemic infection
Characterization of TNF and TNF receptor superfamily in amphioxus
The interferon regulatory factor (IRF) family in amphioxus
The Functional Characterization of Zebrafish Type I Interferon (zfIFN) in Nervous Necrosis Virus
A Multiplex RT-PCR Assay for Rapid and Sensitive Detection of Cytokines in Pufferfish
Earthworm extract possessed anti-inflammatory property through the down-regulation of IL-8 transcription
N-Terminal Region Responsible for Chemotactic Activity of Flounder IL-8
Teleost IL-6 promotes antibody production through STAT3 signaling via IL-6R and gp130
Molecular cloning and characterization of orange-spotted grouper (Epinephelus coioides) IL-12
Gene Knockdown and mRNA Variants of Astakine, Ancient Cytokine, in Kuruma Shrimp Marsupenaeus japonicus
The interferon (IFN) system of salmonids is more complex than previously realised: Characterisation of the IFN locus in rainbow trout
reveals two novel IFN subgroups
Molecular Cloning, Characterization and Expression Analysis of STING from Orange-spotted Grouper, Epinephelus coioides
CLONING AND EXPRESSION ANALYSIS OF A THIRD IL1beta GENE IN SALMONIDS: TWO TYPES OF IL1beta GENES EXISTIN TELEOST FISH

17:10 CCK-O6
17:30 CCK-O7
10:30 CCK-O8
10:50 CCK-O9
11:10 CCK-O10
CCK-P1
CCK-P2
CCK-P3
CCK-P4
CCK-P5
CCK-P6
CCK-P7
Poster
CCK-P8
CCK-P9Y. Ding
Chris Secombes
M. Chadzinska
C. Tafalla
C. Tafalla
Linyi Hu
T. Zheng
Ming-Wei Lu
G. Biswas
T. Iwase
O. Kurata
M. Kaneda
Jui-Ling Tsai
M. Inada

CCK-P10Chris Secombes

Time

CCK-P11

Zhengliang Ou-yang

CCK-P12Chris Secombes

ID No.
10:30 IGR-O1
10:50 IGR-O2
11:10 IGR-O3

Presenter
Y. Sutoh
S. S. Higgie
H. Cortes

11:30 IGR-O4

B.C. Montgomery

11:50 IGR-O5
12:10 IGR-O6
13:30 IGR-O7
13:50 IGR-O8
14:10 IGR-O9
14:30 IGR-O10
14:50 IGR-O11

J. Oriol Sunyer
Katherine A. Rego
Ivar Hordvik
I. Salinas
Nil R. Saha
Gang Cheng
S. D. Fugmann

Title
VLRC defines a population of lamprey lymphocytes distinct from that expressing VLRA or VLRBThe origin and evolution of the immunoglobulin domain
Understanding Channel Catfish (Ictalurus punctatus) leukocyte immune-type receptors (IpLITRs)
Channel catfish leukocyte immune-type receptors mediate inhibition of cellular cytotoxicity by SHP-1-dependent and -independent
mechanisms.
Dominant role of IgT and IgT+ B cells in Rainbow Trout (Oncorhynchus mykiss) cutaneous immunity
Characterization of a new immunoglobulin light chain isotype, Ig lambda, in rainbow trout, Oncorhynchus mykiss
Characterization of IgM sub-variants in salmonid fish
Analysis of immunoglobulin heavy chain genes and proteins in the African lungfish Protopterus dolloi
Genomic survey of the immune repertoire in the living coelacanth
Extensive diversification of IgH subclass encoding genes and IgM to IgM class switching in Crocodylia
Separation of mutational and transcriptional enhancers in the Gallus gallus IGL gene

3/7 ページ

★
★
★
★

★
★

★
★

★

Session Name and Date
Innate cellular immunity (INC)

Day
Wed

15:10 IGR-O12
IGR-P1
IGR-P2
IGR-P3
Poster
IGR-P4
IGR-P5
IGR-P6
IGR-P7

P. Boudinot
M.-S. Wu
S. SudaYoung-Kyu Kim
F. Ramirez-Gomez
Priti Kataria
Tao Wang
Priti Kataria

Teleost fish mount complex clonal IgM and IgT responses in spleen upon systemic viral infection
Expression patterns of the immunoglobulin light chain genes in Epinephelus coioides
Sequencing immunoglobulin variable regions of new antigen receptor genes in the banded houndshark, Triakis scyllium
Characterization of Monoclonal Antibody Against Immunoglobulin Kappa Light Chain in Olive Flounder, Paralichthys olivaceus
Understanding secretory IgD in Rainbow trout: Antigen-specific plasma cell distribution and expression in body secretions
The structure of rainbow trout (Oncorhynchus mykiss) IgM influences antibody-mediated phagocytosis
Evidence of IgY subclass diversification in the reptilian snakes: evolutionary implications 1, 2Competitive displacement of antibody secreting cells in rainbow trout

Time

Presenter
E. Koppang
Tomáš Korytář
L. Pijanowski
Valerie J. Smith
Elisabeth A. Dyrynda
M. A. Kepka
T. Nagasawa
Daniel R. Barreda
Julie Ghosh
H. Miyazaki

INC-P3

Y. Sugie

INC-P4
INC-P5

M. Tomana
Yong-An Zhang

Title
A role of melanin production in melanomacrophages?
Functional and Molecular Aspects of the Immune Defence in Peritoneal Cavity of Rainbow Trout (Oncorhynchus mykiss)
Neutrophil extracellular traps in carp
Extracellular chromatin traps are an ancient eukaryotic cellular defence strategy
Revisiting the cell: Differences in haemocyte death patterns associated with immunity in an invertebrate model
Melatonin reduces ROS-dependent apoptosis of carp phagocytes
Phagocytosis and bactericidal abilities of teleost thrombocytes.
Evolving Roles of Phagocytes for the Control of Inflammation
The axial organ: the source of coelomocytes in the purple sea urchin?
Fatty acid binding protein 7 regulates the functions of murine macrophages.
Carp thrombopoietin in combination with kit ligand a induces the formation of colonies derived from unipotent progenitors forthrombocytes and bipotent progenitors for thrombocytes and erythrocytes
Flow cytometric characterization of Japanese bullhead and whale shark peripheral blood leukocytes
The Neural Cell Adhesion Molecules in Mandarin Fish

ID No.
13:30 IVH-O1
13:50 IVH-O2
14:10 IVH-O3
14:30 IVH-O4
14:50 IVH-O5
15:40 IVH-O6
IVH-P1
IVH-P2
IVH-P3
IVH-P4
Poster
IVH-P5
IVH-P6
IVH-P7
IVH-P8Presenter
V. Skara
B. D. Parsons
P. Prochazkova
C. Noonin
Jiann-Horng Leu
Christine Paillard
A. Accorsi
J. Y. Hwang
M. Kondo
D. Sekine
A. Tame
Irene Söderhäll
A. Watthanasurorot
Valerie J. Smith

Title
Activity of Radix lagotis (Lymnaeidae) hemocytes: influence of infection by the bird schistosome Trichobilharzia regenti
Requirement of Pvr Ligands in Drosophila Embryonic Macrophage Survival and Migration
The regulation of cellular iron homeostasis in Eisenia andrei earthworms
CRAYFISH HEMATOPOIETIC TISSUE AS A MODEL FOR STEM CELL
Identification of hematopoiesis-related genes in Litopenaeus vannamei
Cellular and molecular analysis of Ruditapes philippinarum hemocytes reveals cytoskeleton disruption after Vibrio tapetis challengeTime-course analysis of hemocyte populations in the freshwater snail Pomacea canaliculata (Gastropoda, Ampullariidae)
Flow cytometry analysis hemocytes of immune response of invertebrate due to infections
Morphological Diversity of Crustacean Hemocytes
Monoclonal antibodies recognizing hemocytes and cells in gill tissue of deep-sea mussel, Bathymodiolus japonicus
Comparative morphology of hemocytes in deep-sea symbiotic bivalves, three Bathymodiolus mussels and two Calyptogena clams
Crustacean hematopoietic factor - a regulator of hemocyte homeostasis
Hemocyte homeostasis the role of astakines
Characterisation of prohaemocytes from crab, Carcinus maenas, and their survival in vitro

Time

Presenter
Christine Paillard
C. Le Bris
K. Somboonwiwat
Lijun Feng
R. Bettencourt
Martin Bilej
Han-Ching Wang
Kenneth Söderhäll
Tze Hann Ng
Patrick C. Hanington
Judith E. Humphries

Title
Nacrezation process, an immune defense system efficient against vibrios in manila clams?
Demonstration of the increase of laccase-like activity in immune defense of clams infected by pathogens
Antibacterial and Antiviral Activity of Penaeus monodon Antilipopolysaccaride Factor Isoform 3 (ALFPm3)
Identification and Characterization of an Amphioxus Matrix Metalloproteinase Homolog BbMMPL2 Responding to Bacteria Challenge
The emergence of the mussel Bathymodiolus azoricus as a bona fide model to study innate immunity in deep-sea vent animals
Microbial environment determines the innate immunity in earthworms
The diversified IgSF molecule, Dscam, and highly specific innate immune responses in shrimp
Dscam (Down syndrome adhesion molecule); structure and function in a crustacean.
Investigating Crustacean Immune Response via Putative Antigen-Specific Receptor, Dscam
FREP3 is an important factor in snail resistance to trematode infection
Gene regulation of immune responses in Biomphalaria glabrataThu

ID No.
11:15 INC-O1
11:35 INC-O3
10:30 INC-O4
10:50 INC-O7
11:10 INC-O6
11:30 INC-O2
11:50 INC-O5
12:10 INC-O3
INC-P1
INC-P2

PosterSession Name and Date
Invertebrate hemocytes (IVH)

Session Name and Date
Invertebrate immune response (INV)

Day
Thu

Day
Thu

Fri

Time

ID No.
16:00 INV-O1
16:20 INV-O2
16:40 INV-O3
17:00 INV-O4
17:20 INV-O5
10:30 INV-O6
10:50 INV-O7
11:10 INV-O8
11:30 INV-O9
11:50 INV-O10
12:10 INV-O11

4/7 ページ

★

★

★
★

★
★

13:30 INV-O12
13:50 INV-O13
14:10 INV-O14
14:30 INV-O15
15:20 INV-O16
15:40 INV-O17
16:00 INV-O18
INV-P1
INV-P2
INV-P3
INV-P4

F.Y. Chen

INV-P5

K QiaoINV-P6
INV-P7
INV-P8
INV-P9INV-P10
INV-P11
INV-P12
INV-P13

INV-P15
INV-P16
INV-P17

S.L. Hsieh
T. Itami
P. Jiravanichpaisal
Jiann-Chu Chen
M. Inada
Tohru Mekata
Taku Hibino
Chadanat Noonin
Francisco RamirezGomez
Hiroki Inoue
Christopher J. Bayne
S. Dharaneedharan

INV-P18

T. Itami

INV-P19
INV-P20
INV-P21
INV-P22

T. Itami
Xin Tao
H. KanohYong-Chin Lin

ID No.
14:30 LEC-O1
14:50 LEC-O2

Presenter
P. Angthong
Laura F. Grice

15:10 LEC-O3

Lijun Feng

Poster

INV-P14

Session Name and Date
Lectin (LEC)

Session Name and Date
Pattern recognition molecules (PRM)

Day
Tue

Day
Tue

Time

C. H. Bigger
S. Guntermann
T. Shibata
L. Zhang
Edan Foley
C.J. Coates
Moon-Soo Heo
J. Peng
X. Dong
M. Tsau

16:00 LEC-O4
16:20 LEC-O5
LEC-P1
Poster
LEC-P2
LEC-P3

Maria Forlenza
A.M. Argayosa
Y. Ikazaki
N. Wakamiya
Y. EndoTime

ID No.
10:30 PRM-O1
10:50 PRM-O2

Presenter
S.V NairM. O Roth

11:10 PRM-O3

C. M. Lun

11:30 PRM-O4
11:50 PRM-O5

T.P. Yoshino
Nil R. Saha

12:10 PRM-O6

Danilo Pietretti

Adaptive Immunity in a Coral: a Question of Memory?
Functional Analysis of the Fly Caspase Dredd in the IMD Innate Immune Signaling Pathway.
Transglutaminase-catalyzed Relish Crosslinking Suppresses Innate Immune Signaling in the Drosophila Gut
Evolutionary and Regulation Dynamics of Immune-Related Genes and Pathways in the Pacific Oyster
The Drosophila Pvr Pathway is an Intrinsic Autocrine Regulator of Intestinal Homeostasis.
Investigating Amebocyte Phagocytosis and Immune Defence in Limulus polyphemus
Immunomodulation of green marine alga (Ulva lactuca) against Aeromonas hydrophila in Macrobrachium nobilii
Novel TIR adaptors acts as a negative regulator in inflammatory signaling of amphioxus
Ubiquitination regulatory of immune signaling pathway in amphioxus
Cloning and analysis of autophagy-related protein 8 (ATG8) in shrimp Litopenaeus vannamei
Gene cloning gene protein expression profiling and antioxidant activity of some immune associated parameters in Scylla paramamosain
with LPS challenge
Gene cloning and expression analysis of a sigma class glutathione S transferase and molluscan caspase in haemocytes of variously
colored abalone upon bacterial challenge
Effects of Dietary Gynura bicolor Extract on Immune Responses in White Shrimp, Litopenaeus vannamei
Molecular Cloning and Analysis of Toll Interacting Protein Gene, MjTollip, in Kuruma Shrimp Marsupenaeus japonicus
Properties of an Interleukin-1 Receptor-Associated Kinase-4 (IRAK-4) of Black Tiger Shrimp (Penaeus monodon)
Modulation of the innate immune response in white shrimp following long-term low-salinity exposure
Molecular Cloning and Expression Analyses of Free Radical Generation Enzymes in Kuruma Shrimp Marsupenaeus japonicus
Changes in transcription profiles of immune related genes after silencing of Toll receptors in shrimp
Slow transport of foreign objects to the distal end of the sea urchin larval arms
Identification and functional study of thymosins in Pacifastacus leniousculus★

★

Highlights from cellular and molecular studies of the immune system of a Caribbean Sea cucumber
Characterizations of Novel Cadherins and Intercellular Recognition Molecules Related to Parasite-Host Compatible Cell Interactions.
Resistance to Schistosoma mansoni is correlated with the numbers of pseudopod-producing hemocytes in Biomphalaria glabrataMagnifera indica kernel feed on immune response of Penaeus indicus against white spot syndrome virus (WSSV)Interaction between WSSV infection and Apoptosis-, JAK/STAT-signaling- pathway- and RNAi-related Genes in Kuruma Shrimp,
Marsupenaeus japonicus
Identification and Expression Analysis of Cytochrome c in Kuruma Shrimp, Marsupenaeus japonicus
Amphioxus IKK complex involved in TLR modulated NF-kappaB and IRF activation
Characterization of a downstream component of Drosophila receptor guanylate cyclase activating innate immune response
The Antioxidant Role of Superoxide Dismutase in Penaeid Shrimp
Title
Characterization of the Tachylectin-type Lectin in the Black Tiger Shrimp, Penaeus monodon
Seeking diversity: Genomic characterisation of the aggregation factor genes of the demosponge Amphimedon queenslandica
Identification of an amphioxus intelectin homologue that preferably agglutinates gram positive over negative bacteria likely due todifferent binding capacity to LPS and PGN
Analysis of Highly Expressed Short Immune-Related Soluble C-Type Lectin (SISC) in Common Carp (Cyprinus carpio)
Warm-water Fish Lectins and Their Relevance to Innate Immunity and Aquaculture Applications
Possible roles of Xenopus laevis XCGL family lectins in innate immunity
The Biological Functions of the Novel Collectins CL-L1, CL-K1, and CL-P1
Orthology between human and mouse ficolins, and roles of mouse ficolins A and B in host defense
Title
Variations to the sea urchin 185/333 genes during gametogenesis and ontogeny
Somatic gene diversification in the sea urchin, Heliocidaris erythrogramma
A recombinant Sp185/333 immune protein from the California purple sea urchin shows specific and non-reversible binding to certain
microbes
Innate immunity in snails Biomphalaria glabrata to schistosome infections: early recognition based on lectin-glycan interactions
Extreme Functional Evolutionary Divergence of an Immune Receptor in a Basal Vertebrate
POSSIBLE ROLE IN RECOGNITION OF PROTOZOAN PARASITES: CHARACTERIZATION OF TLR20 IN COMMON CARP(CYPRINUS CARPIO)

5/7 ページ★

13:30 PRM-O7
13:50 PRM-O8
14:10 PRM-O9

Pinwen P. Chiou
C.L. Yao
Inge R. Fink

PRM-P1

Ming-Ching Lin

PRM-P2

M.X. Chang

PRM-P3
PRM-P4

H. Chou
Lauren S. Sherman

PRM-P5

T. Aoki

PRM-P6
PRM-P7
PRM-P8
PRM-P9PRM-P10Danilo Pietretti
Danilo Pietretti
Seong Don Hwang
R. Harikrishnan
SeongDon Hwang

PosterSession Name and Date
T-cell, MHC, and Ag-presentation (TCM)

Day
Fri

Time

ID No.
11:10 TCM-O1
11:30 TCM-O2
11:50 TCM-O3
12:10 TCM-O4
13:30 TCM-O5
13:50 TCM-O6
14:10 TCM-O7
14:30 TCM-O8
15:20 TCM-O9
15:40 TCM-O10

Presenter
N.T Fujito
C. Huang
H. Bannai
Uwe Fischer
M.F. CriscitielloG. KatoG. Scapigliati
Huong Dang Thi
T. Yamaguchi
Yuko Ota

16:00 TCM-O11

David Parra

16:20 TCM-O12
TCM-P1
TCM-P2
TCM-P3
TCM-P4
TCM-P5
Poster
TCM-P6
TCM-P7

Session Name and Date
Vaccine and adjuvant (VAC)

Day
Wed

Time

Fumio Takizawa
S. Yoon
P. Johansson
Y. YanM. I. Nonaka
R. Nagamine
K. Araki
Jorunn B. Jørgensen

TCM-P8Chun Xia

TCM-P9

Fumio Takizawa

ID No.

Presenter

10:15 VAC-O1

H. L. Thim

10:35 VAC-O2
10:55 VAC-O3
11:15 VAC-O4
11:35 VAC-O5
11:55 VAC-O6
VAC-P1

K. Villumsen
L. Neumann
M. K. Raida
L. G. Jørgensen
Dimitar Iliev
Dong-Hwi Kim

Poster

VAC-P2

M.-G. Kwon

VAC-P3

A. Taechavasonyoo

Regulation of Grouper Toll-like Receptor 9 Signaling by Alternative RNA Splicing
Molecular characterizations of two types of TLR5 in large yellow croaker, Larimichthys crocea
Studies into the Formation of Heterodimers of Toll-Like Receptor-1 and -2 in Common Carp
The shrimp anti-lipopolysaccharide factor (SALF) inhibits the expression of proinflammatory cytokines by Trichomonas vaginalis infected HeLa cells.
Identification of peptidoglycan recognition protein 6 and its splicing isoforms that inhibit intracellular bacterial proliferation in grass carp
CIK cells
Does opsonization of bacteria by a recombinant Sp185/333 protein from the California purple sea urchin augment phagocytosis?
Diversity of immune response proteins: nickel-isolated Sp185/333 proteins from the purple sea urchin
Evolutional Conservation of Primordial Functions and Transcriptional Control of LGP2 Gene in Japanese Flounder, Paralichthys★
olivaceus
CHARACTERIZATION OF TOLL-LIKE RECEPTOR 4 IN COMMON CARP (CYPRINUS CARPIO)
Identification of several CRP-like sequences in common carp increases the complexity of the pentraxin system
Molecular identification and expression analysis of two distinct BPI/LBPs from rock bream, Oplegnathus fasciatus
Identification and molecular characterization and mRNA transcription in tissues of the apolipoprotein E in Epinephelus bruneus
Molecular cloning and functional characterization of Toll-like receptor 3 in Japanese flounder, Paralichthys olivaceus
Title
Long-term Transspecies Dimorphism of the PSMB8 Gene in the Actinopterygian MHC Region
Recurrent Revival of Dimorphism of the PSMB8 Gene During Tetrapod Evolution
Evolution of Teleost MHC Class II Genes Revealed by Comprehensive Analysis of Medaka
Phenotypic differences of mucosal and non-mucosal CD8alpha positive T-cells in the teleost Oncorhynchus mykiss
Somatic hypermutation for primary alpha/beta T cell repertoire generation in shark
T cell marker CD4 and CD8 gene in Japanese flounder, Paralichthys olivaceus
T cells of the teleost sea bass Dicentrarchus labrax (L.)
A profile of peripheral and mucosal T-cell subpopulations in rainbow trout (Oncorhynchus mykiss)
Clonal culture of carp Th2-like cells whose immune status is hallmarked by IL-4/13B gene expression
Evolution of the B7 family coevolution and the B7s historical relationship with the MHC
Phagocytic B cells discovered in mammals: Murine B-1 B cells have phagocytic and microbicidal capacities and present phagocytosed
antigen to CD4+ T cells
First identification of regulatory B cell subsets expressing IL-10 in teleost fish
CHARACTERIZING THE ROLE OF CD4+ CELLS WITHIN THE ZEBRAFISH (DANIO RERIO) IMMUNE RESPONSE
Characterising putative dendritic cell markers in rainbow trout (Oncorhynchus mykiss)
Functional studies on an immune-related gene of MyD88 in orange-spotted grouper, Epinephelus coioides
Different Evolutionary Patterns of Three Alpha Domains of the MHC Class IA Genes
Characterization and expression analysis of the Japanese pufferfish Takifugu rubripes homologue of the mammalian Th-POK
Characterization and expression analysis of CD4 and CD8 genes in yellowtail, Seriola quinqueradiata
CD40L An important Costimulatory molecule involved in the maturation of Antigen Presenting Cells in Atlantic salmon.
Narrow Groove of MHC Class I Molecule BF2*0401 Plus Peptide Transporter Restriction can Explain Disease Susceptibility of B4
Chickens
Identification and transcription analysis of Eomesodemin-a and -b in rainbow trout
Title
INCREASED PROTECTIVE IMMUNE RESPONSES WHEN COMBINING TLR-AGONISTS IN A WHOLE-VIRUS ANTIGEN
VACCINE AGAINST SALMONID ALPHAVIRUS
Protective effects of vaccine-induced specific antibodies against Aeromonas salmonicida subsp. salmonicida in rainbow trout
Oral and anal vaccination against enteric red mouth disease induces full protection against yersiniosis.
Immersion vaccination against Yersinia ruckeri O1, biotype 2 confers cross protection against Y. ruckeri O1 biotype 1.
Improved ERM vaccination efficacy using combined vaccine administration methods
CpG activation of distinct MHCII+ cell types in Atlantic salmon (Salmo salar)
Effects of chitin and chitosan particles on innate immune system in Epinephelus bruneus
The efficacy and safety of oil-adjuvanted combination vaccine containing Edwardsiella tarda, Streptococcus iniae and Streptococcus
parauberis in olive flounder Paralichthys olivaceus
The immuno-adjuvant effect of interleukin-1 beta in Japanese flounder (Paralichthys olivaceus)

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VAC-P4
VAC-P5
Session Name and Date
Vertebrate adaptive immune response (ADP)

Day
Thu

Time

ID No.
10:30 ADP-O1
10:50 ADP-O2
11:10 ADP-O3
11:30 ADP-O4
11:50 ADP-O5
12:10 ADP-O6
13:30 ADP-O7
13:50 ADP-O8
14:10 ADP-O9
ADP-P1
ADP-P2
ADP-P3

Poster

ADP-P4
ADP-P5ADP-P7
ADP-P8ADP-P9Session Name and Date
Vertebrate innate/homeostatic response (VIH)

Day
Thu

Fri

Time

S. Chen
Bo-Wei Chi

Vaccine efficacy of recombinant proteins from Nocardia seriolae against nocardiosis in large mouth bass, Micropterus salmoides
Development of oral delivery system for aquaculture viral disease using Listeria monocytogenes

Presenter
Makoto Inami
M. K. Raida
M. Yamasaki

Title
Intestinal immunity in cod
Association between increased antibody level and protection in Yersinia ruckeri bacterin immersion vaccinated rainbow trout.
Cell-mediated and humoral immune response to Edwardsiella tarda in ginbuna crucian carp, Carassius auratus langsdorfii
Protection of ginbuna crucian carp (Carassius auratus langsdorfii) against Edwardsiella tarda infection using adoptive transfer of
K. Arakicytotoxic T lymphocytes
K. Ohishi
Three dimensional models of Signaling Lymphocyte Activation Molecules (SLAM), a receptor for morbillivirus in carnivores
M. Edwards
Immune protection of developing marsupial pouch young: what does transcriptome sequencing tell us?
L.Jørgensen
Immunity against the parasite Ichthyophthirius multifiliis in rainbow trout
Louise A. Rollins-Smith Deadly chytrid fungus can paralyze amphibian lymphocyte responses
T. Odaka
The Plasmablast-like Leukocyte in the Kidney of Fugu (Takifugu rubripes)
Seung-Hyun Hong
Dietary Rubus coreanus on immune system in Epinephelus bruneus against Vibrio alginolyticus
Ronggai Li
B-cell markers and cytokines involved in B-cell growth and differentiation from cartilaginous fish
S. Boltaña
Immune response in sea bream (Sparus aurata) after treatment with LPS of Aeromonas salmonicida and Listonella anguillarum
MOLECULAR CHARACTERIZATION OF IMMUNE RESPONSES ELICITED DURING PKD INFECTION IN BROWN TROUT vs
B. Gorgoglione
RAINBOW TROUT
Chun-Hsin Wu
Computational Analysis of Immune Foods and Diets Based on Nutritional Similarity Measurement
Haruka Tsukamoto
Characterization of culture supernatant of Streptococcus parauberis: hemolytic factor and leucocyte-stimulation in Japanese flounderJ. W. Hodgkinson
Analysis of the immune interface of Mycobacterium marinum infections in the goldfish (Carassius auratus L.)
M. Zhang
Depletion of conventional mature B cells and compromised specific antibody response in bovine Ig mu heavy chain transgenic mice

ID No.
14:30 VIH-O1
14:50 VIH-O2
15:40 VIH-O3

Presenter
S.MacKenzie
A. Frøyse
Whitney M. Gammill

16:00 VIH-O4

Dominika A. Przybylska Beta-glucan enriched bath directly stimulates the wound healing process in common carp (Cyprinus carpio L.)

16:20 VIH-O5
16:40 VIH-O6
17:00 VIH-O7
17:20 VIH-O8
10:30 VIH-O9
10:50 VIH-O10
VIH-P1
VIH-P2
VIH-P3
VIH-P5
Poster
VIH-P6
VIH-P7
VIH-P8
VIH-P9T. Wulff
J.G.Schmidt
N.I. Vera-Jiménez
N.I. Vera-Jiménez
J Hikima
A. Skjesol
R. Belmonte
J. WeiS. Tasumi
J. KangSeong Don HwangSònia Rey Planellas
Josep V. Planas
A. Callol

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Title
Behavioural fever is a synergic signal amplifying the innate immune response
Inflammation - a co- factor in salmonid coronary disease?
Innate immune defenses in amphibian skin against the fungal pathogen Batrachochytrium dendrobatidis

Immunological aspects of tissue regeneration in rainbow trout muscle - A proteomics and bioinformatics approach
Innate immune responses to experimentally inflicted wounds in common carp (Cyprinus carpio L.) larvae and juveniles
Carp head-kidney leukocytes display different oxygen radical production patterns after DAMP and PAMP stimulation.
Modulation of fish fibroblast proliferation with B-glucan and hydrogen peroxide during scratch-wound healing in vitro.
Expression of Type-I Interferon Gene Controlled by IRF3/7 Mediated through TBK1 in Japanese Flounder
Suppressors of Cytokine Signaling (SOCS) in Atlantic Salmon
The Immune Response of Atlantic Salmon to Saprolegnia
Cloning, characterization, and expression analysis of a thioredoxin from orange-spotted grouper (Epinephelus coioides)
Candidate key molecule(s) determining host specificity of parasite on fugu, Takifugu rubripes
Skewed segregation of a microsatellite marker linked with homozyous lethality after metamorphose of olive flounder
Molecular characterization, expression, and functional analysis of two thioredoxins in the black rockfish (Sebastes schlegelii)
Emotional fever in the zebrafish (Danio rerio) during intra-peritoneal injection procedures.
Modulation of the transcriptomic effects of lipopolysaccharide in skeletal muscle by swimming in zebrafish (Danio rerio)
European eel (Anguilla anguilla) 454 immunotrascriptome sequencing.

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