WPI Immunology Frontier Research Center FY 2013

Edit : Research Planning & Management Office, IFReC
Published in June, 2014

O s a k a U n i ve r s i t y

Copyright : Immunology Frontier Research Center (IFReC ), Osaka University

World Premier International
Research Center

Osaka University

Immunology
Frontier
ResearchCenterAnnual Reportof IFReC
FY 2013

World Premier International
Research Center

Message from the Director

Osaka University

Immunology
Frontier
ResearchCenterAs the Director of the Immunology Frontier
Research Center (WPI-IFReC) at Osaka University,
I am very pleased to present the IFReC annual
report for fiscal 2013.
Since its inception in 2007, IFReC has established itself as a Visible International ResearchCenter of Immunology with the support of many
people including the WPI Program Committee,
the Program Director and the Program Officers.

Contents
Message from the Director ...................................................

1

Science Talk Live 2013 by WPI ............................................. 83

Organization Chart ..................................................................

2

Symposium for General Audience ..................................... 84

Committee and Advisory Board for IFReC .......................

4

AAAS 2014 .................................................................................. 85

Administrative Office of IFReC .............................................

6

Research Projects ........................................................................ 87

Laboratories ..................................................................................

7

Immunology Group .................................................................

8

Research Support Program for
Fusion of Different Fields ...................................................... 88

Imaging Group .......................................................................... 38
Informatics Group .................................................................... 52
Units for Combined Research Fields ................................. 56
Symposia & Seminars ............................................................... 61
International Symposium TCUID ........................................ 62
Immunology Frontier "B to B Seminar" ............................ 64
Mini Symposium on Malaria Immunopathology
Kishimoto Foundation Lecture ........................................... 65
IFReC Seminars .......................................................................... 66
IFReC Colloquia ......................................................................... 68
Live Immuno-Imaging Facility Opening Workshop .... 70
Events ............................................................................................... 71
Winter School ............................................................................ 72
Staff and Researchers Development Seminars ............. 74
Japanese Lessons ..................................................................... 76
Outreach Activities .................................................................... 79
Science Café Series “Café on the Edge” ............................ 80
Super Science High School Festa 2013 ............................ 81
Science Agora 2013 ................................................................. 82

Combined Research Fields Evaluation Workshop ........ 90
Dual Mentor Program ............................................................. 91
Young Scientist Support Program for Research Abroad
FIRST Program : AKIRA Project ............................................. 92
Data ................................................................................................... 97
Facilities ....................................................................................... 98
Kishimoto Foundation Fellowships ................................... 100
Members ..................................................................................... 101
Awards ......................................................................................... 102
Finance ......................................................................................... 104

In the WPI interim evaluation carried out at theend of fiscal year 2011, IFReC was awarded ascore of ‘A’ and it was noted that IFReC should bemade a permanent part of the University as an
International Level Research Institute. In alignment with this view, we are currently finalizing
the future direction of IFReC at Osaka University
with the University Executive Board as well as the
Ministry of Education, Culture, Sports, Scienceand Technology (MEXT). We expect that an
important decision will be made as to thecontinuation of IFReC beyond 2017 at the WPI
program committee meeting to be held in fall2014.

As an approach to nurturing young researchers, the third NIF Winter School, which was
begun in 2011, was held in January, 2014 on Awaji
Island. The young researchers, selected from ahighly competitive field of candidates, spent one
week living and debating together in an‘Immunology Boot Camp’ that is fast becoming
an institution of its own. The school prides itself
on offering not only productive educational
content but also an opportunity for the young
researchers to form a global network.
We are committed to continuing contributions to scientific advances through immunology
research and education and the evolvement of a
Visible Research Center that leads the world.

To this end, IFReC must also demonstrate its
worth as a World Premier Research Institute.
However, it is not sufficient to simply maintainthe level of research. IFReC has been running an
original research grant ‘Research Support
Program for Combined Research Fields’ in orderto promote collaborative studies between different fields. In 2013, two Combined Research Units
were set up to advance this program. Both units
comprise a group of young, talented researchers
from different fields and we expect that lively
debate will pave the way to new areas ofresearch.

Research Outputs ....................................................................... 105
Selected Articles ....................................................................... 106
Publications ................................................................................ 110
Lectures by PIs ........................................................................... 116
Access Map ..................................................................................... 118

Shizuo Akira, MD/PhD
Director
WPI Immunology Frontier
Research Center

1Immunology Group

Organization Chart

MEXT
WPI Initiative
Program Committee

JSPS

WPI Working GroupInternational
Scientific
Advisory Board

Host Defense ......................................................................................Shizuo Akira
Immunoglycobiology .....................................................................Taroh Kinoshita
Immunopathology ..........................................................................Atsushi Kumanogoh
Immunochemistry ............................................................................Hisashi Arase
Immune Regulation .........................................................................Tadamitsu Kishimoto
Mucosal Immunology .....................................................................Kiyoshi Takeda
Molecular Immunology .................................................................Hitoshi Kikutani
Experimental Immunology ...........................................................Shimon Sakaguchi
Cell Signaling ......................................................................................Takashi Saito
Lymphocyte Differentiation .........................................................Tomohiro Kurosaki
Lymphocyte Development ...........................................................Fritz Melchers
Malaria Immunology .......................................................................Cevayir Coban
Vaccine Science .................................................................................Ken J. Ishii
Immune Regulation .........................................................................Tsuneyasu Kaisho
Immune Network .............................................................................Rikinari Hanayama
Immunoparasitology ......................................................................Masahiro Yamamoto

Imaging Group

Director

Research Groups

Deputy Directors

Management
Committee

Board ofRepresentatives

Administrative
Director
General Affairs Section

Accounting Section

Single Molecule Imaging ...............................................................Toshio Yanagida
Biofunctional Imaging ....................................................................Yoshichika Yoshioka
Immunology and Cell Biology .....................................................Masaru Ishii
Nuclear Medicine .............................................................................Jun Hatazawa
Biophotonics ......................................................................................Nicholas Isaac Smith
Chemical Imaging Techniques ....................................................Kazuya Kikuchi
Immune Response Dynamics ......................................................Kazuhiro Suzuki

Informatics Group
Information Systems .......................................................................Yutaka Hata
Systems Immunology .....................................................................Daron M. Standley

Units for Combined Research Fields

Administrative Department

Quantitative Immunology Research Unit ................................Yutaro Kumagai,
Shunsuke Teraguchi, Diego Diez
Next Generation Optical Immune-imaging ............................Noriko Takegahara,
Kazuaki Tokunaga

Liaison Office

Research PlanningandManagement Office

Common Facility

Osaka University
President
Administrative Bureau

2Core Instrumentation Facility
Animal Resource Center for Infectious Diseases
Network Administration OfficeInstitute for Frontier Medical Sciences, Kyoto University

Domestic RIKEN Center for Integrative Medical Sciences
National Institute of Biomedical Innovation

Cooperative
Institutions
Overseas

Pohang University of Science and Technology, Korea
Convergent Research Consortium for Immunologic Disease,
Seoul St. Mary's Hospital, the Catholic University of Korea
Indian Institute of Science Education and Research, India
Maurice Wilkins Center, The University of Auckland, New Zealand3

Committee and Advisory Board for IFReC
World Premier International Research Center Initiative (WPI)
Program Committee Members List

As at April, 2013

<Chair>Hiroo Imura

President, Foundation for Biomedical Research and Innovation

Toshiaki Ikoma

Executive Vice President & CTO, Canon Inc.

Hiroto Ishida

President Emeritus, Kanazawa Gakuin University

Shinichiro Ohgaki

Professor Emeritus, The University of Tokyo

Tsutomu Kimura

Chairman, Tokyo Metropolitan Government Board of Education

Kiyoshi Kurokawa

Academic Fellow, National Graduate Institute for Policy Studies

Makoto Kobayashi

Director, Research Center for Science Systems, Japan Society for the Promotion of Science (JSPS),
Nobel Laureate in Physics (2008)

Masatoshi Takeichi

Director, RIKEN, Kobe InstituteMichiharu Nakamura

As at April, 2013

Jeffrey Ravetch

Professor, Laboratory of Molecular Genetics and Immunology, The Rockefeller
University

Immunology

Chris Goodnow

Professor, John Curtin School of Medical Research and Australian Phonemics
Facility, The Australian National University

Immunology

Richard Locksley

Professor, Departments of Medicine and Microbiology/ Immunology, University ofCalifornia, San Francisco

Immunology

Anne O'Garra

Division Head, Division of Immunoregulation, The National Institute for Medical
Research

Immunology

Lewis LanierAmerican Cancer Society Professor Chair, Department of Microbiology &Immunology, University of California, San Francisco

Immunology

Kiyoshi Takatsu

Director, Toyama Prefectural Institute for Pharmaceutical Research

Immunology

President, Japan Science and Technology Agency(JST)Kayo Inaba

Professor, Graduate School of Biostudies, Kyoto University

Immunology

Ryoji Noyori

President, RIKEN, Nobel Laureate in Chemistry (2001)

Yasuyoshi Watanabe

Director, CLST, RIKEN Kobe InstituteImaging

Hideo Miyahara

Professor Emeritus, Osaka University

Masamitsu Iino

Professor, Graduate School of Medicine, The University of Tokyo

Imaging

Robert Aymar

Senior Counsellor, “Administrateur Général”(CEO), French Atomic Energy Authority(CEA)

Rita Colwell

Professor, University of Maryland

Yale Goldman

Professor, Pennsylvania Muscle Institute, University of Pennsylvania

Imaging

Richard Dasher

Professor, Stanford University

Akinori Kidera

Professor, Graduate School of Integrated Science, Yokohama City University

Informatics

Ian Halliday

Professor Emeritus, University of Edinburgh

Hiroyuki Toh

Deputy Director, AIST-CBRC

Informatics

Chuan Poh Lim

Chairman, Agency for Science, Technology and Research, Singapore

Matthew Mason

Director, Robotics Institute, Carnegie Mellon University

David WestheadProfessor, School of Biochemistry and Molecular Biology, Leeds University

Informatics

Vladimir Brusic

Principal Associate, Dana-Farber Cancer Institute, Harvard Medical School

Informatics

Mo Jamshidi

Lutcher Brown Endowed Chair and Professor, Department of Electrical andComputer Engineering, University of Texas, San Antonio

Informatics

Philip Chen

Professor, Faculty of Science and Technology, University of Macau

Informatics

Takeshi Yamakawa

Board Vice-Chairman and Director, Fuzzy Logic Systems Institute

Informatics

Program Director
Toshio Kuroki

Senior Advisor, Research Center for Science Systems (JSPS)

Working Group Leaders and Assigned Members

As at April, 2013

<Chair, Program Officer>
Takehiko Sasazuki

University Professor, Institute for Advanced Study, Kyushu University

Hiroshi Kiyono

Dean and Professor, Division of Mucosal Immunology, Department of Microbiology and
Immunology, Institute of Medical Science, The University of Tokyo

Nagahiro Minato

Dean and Professor, Graduate School of Medicine, Kyoto University

Kazuhiko Yamamoto
Günter J. Hämmerling
Hisataka Kobayashi
Philippe Kourilsky
Diane Mathis

4International Advisory Board

Professor and Chairman, Department of Allergy and Rheumatology, Graduate School of
Medicine, The University of Tokyo
Professor and Chairman, Division of the Molecular Immunology, German Cancer Research Center
Heidelberg DKFZ, DEU
Associate (Chief ) Scientist, Molecular Imaging Program, National Cancer Institute, National
Institutes of Health, USA
Professor, Collège de France/ Honorary Director-General, The Institute of Pasteur/ Chairman, The
Singapore Immunology Network, FR
Morton Grove-Rasmussen Professor of Immunohematology/ Head, Division of Immunology,
Department of Microbiology and Immunobiology, Harvard Medical School, Boston, USA

The WPI Program Committee conducts follow-up activities on progress being made by the WPI institutes with an eye todeveloping them into “highly visible research centers”. They carry out the interim evaluation in close cooperation with a
Program Director, Program Officer and the working group of WPI established to evaluate IFReC. The working group conductssite visits and compiles the results in a Site Visit Report, which is used by the Program Committee along with the results of ahearing on each center to prepare the interim evaluation.
IFReC Advisory Board Members evaluate scientific activities by reviewing PI’s scientific achievements.

5Administrative Office of IFReC
General Affairs Section
 Employment/acceptance of researchers and staff procedures
 Support for international researchers Social insurance/Employment insurance Public dormitory for Osaka University workers
 Procedures related to international students
 Management of work hours

Liaison Office General support for researchers from overseas
 Assistance for each international PI’s lab Providing support for development of an international research
environment Procedures related to patents
 Issuing various certifications

Accounting Section
 Budget drafting/implementation/management
 Purchasing procedures
 Acceptance and implementation of external funding Payment of payroll, travel expenses and honorariums
 Health insurance procedures
 Management of buildings and assets
 RI procedures

Research Planning & Management Office Research Promotion & Support(Consultation for grants & patents, Fusion research program, etc.)
 Establishing Research Environments(Facility & Safety management, Research agreement, etc.)
 Fostering Young Scientists(Winter School, Dual Mentor Program, etc.)
 Organizing Scientific Events(Symposia, Colloquia, Seminars, etc.)
 Public Relations(Publishing, Website, Outreach to citizens, etc.)
 WPI evaluation issues(Progress report, Advisory Board meeting, etc.)

6

Host Defense
The immune system plays a critical role in host defense against microbes. Pattern-recognition receptors (PRRs) sense microbes ranging
from bacteria to fungi, protozoa and viruses, and induce innate immune response, an evolutionally conserved host defense response.
After sensing microbial components, PRRs stimulate productionof inflammatory factors such as cytokines/interferons by dendritic
cells to support induction of acquired immune response, anotherimmune response mediated by T cells and B cells, resulting in elimination of invading microbes. However, aberrant activation of theimmune responses often causes massive inflammation, leading to
the development of autoimmune diseases. Therefore, both activationand inactivation of immune responses must be strictly controlled.
To gain a deeper understanding of the immune system, we have examined regulatory mechanisms of innate and acquired immune responses, and novel PRRs capable of sensing microbial components.

RNA-sensing PRR that induces elimination of intracellular RNA
viruses including MLV.

3. Malt1-dependent cleavage of Regnase-1 regulates
immune response of CD4+ T Cells
Regnase-1 is an RNase that destabilizes a set of mRNAs
through cleavage of their 3' UTRs. Although loss of Regnase-1
leads to development of an autoimmune disease characterized
by T cell activation, one role of Regnase-1 in regulation of T
cell-mediated immunity has remained unclear. We found that
Regnase-1 regulates the mRNAs of a set of genes, including
c-Rel, Ox40, and Il2, through cleavage of their 3' UTRs, and isessential for preventing aberrant cell autonomous generationof effector CD4+ T cells (Uehata et al, Cell, 2013). Regnase-1
is cleaved by Malt1 after T cell receptor (TCR) stimulationand enhances T cell activation. Consistently, Malt1 proteaseactivity is involved in control of the mRNA stability of T celleffector genes. These findings indicate that dynamic controlof Regnase-1 expression is critical for modulation of T cell
activation (Figure 2).
In the future, we will further our research to achieve the goalof a comprehensive understanding of the immune systemand to develop an effective treatment for immune-related
inflammatory diseases.

1. Cathepsin D potentiates poly IC-induced innate immune
response
Shizuo Akira, MD/PhD

RIG-I-like receptors (RLRs) sense viral dsRNA to exert the antiviral
immune response and mediate the adjuvant effect of polyinosinicpolycytidylic acid (poly IC), a synthetic dsRNA. We revealed the regulatory
mechanisms of immune response induced by poly IC (Zou et al.,Immunity, 2013). Poly IC is taken up by dendritic cells (DCs), and induces
lysosomal destabilization, which in turn activates an RLR-dependent
signaling pathway. Upon poly IC stimulation, cathepsin D is released
into the cytoplasm from the lysosome to interact with IPS-1, an adaptor
molecule for RLRs. This interaction facilitates cathepsin D-dependentcleavage of caspase-8 and the activation of NF-κB, resulting in enhanced
cytokine production. Further recruitment of RIP-1 to this complex
initiates the necroptosis of a small number of DCs. HMGB1 released bydying cells enhances IFN-β production in concert with poly IC. Thesefindings indicate that cathepsin D-triggered necroptosis is a mechanismthat propagates the adjuvant efficacy of poly IC (Figure 1).

2. ZAP mediates host defense against murine leukemia
virus

8

Professor

Shizuo Akira

Associate Professor

Tatsuya Saitoh

Assistant Professor

Takashi SatohPostdoctoral Fellow

5

Research Assistant

9

Visiting Scientist

4Support Staff

1

When host cells are infected by an RNA virus, PRRs recognize the viral
RNA and induce the antiviral immune response. However, the PRR thatrecognizes murine leukemia virus (MLV) is not fully understood. We
found that zinc-finger antiviral protein (ZAP) acts as an RNA sensor andinduces the degradation of the MLV transcripts by the exosome, an RNA
degradation system, on RNA granules (Lee et al, PNAS, 2013). The loss
of ZAP greatly enhances the replication efficiency of MLV. ZAP localizes
to RNA granules, where the processing-body proteins assemble. ZAP
induces the recruitment of the MLV transcripts and exosome components
to the RNA granules. The CCCH-type zinc-finger domains of ZAP, which
are RNA-binding motifs, mediate its localization to RNA granules and
MLV transcripts degradation by the exosome. Thus, ZAP is the cytosolic

Figure 1. Poly IC potently activates dendritic cells.

Figure 2. Regnase-1 controls T cell response.

Recent Publications
 Zou J, Kawai T, Tsuchida T, Kozaki T, Tanaka H, Shin KS, Kumar H, Akira S.
Poly IC triggers a cathepsin D- and IPS-1-dependent pathway to enhancecytokine production and mediate dendritic cell necroptosis. Immunity
38:717-28, 2013.
 Uehata T, Iwasaki H, Vandenbon A, Matsushita K, Hernandez-Cuellar E,
Kuniyoshi K, Satoh T, Mino T, Suzuki Y, Standley DM, Tsujimura T, Rakugi H,
Isaka Y, Takeuchi O, Akira S. Malt1-induced cleavage of regnase-1 in CD4+
helper T cells regulates immune activation. Cell 153:1036-49, 2013.

 Maruyama K, Uematsu S, Kondo T, Takeuchi O, Martino MM, Kawasaki T,
Akira S. Strawberry notch homologue 2 regulates osteoclast fusion byenhancing the expression of DC-STAMP. J. Exp. Med. 210:1947-60, 2013.
 Akira S, Misawa T, Satoh T, Saitoh T. Macrophages control innate
inflammation. Diabetes Obes Metab. 15 Suppl. 3:10-18, 2013.

 Lee H, Komano J, Saitoh Y, Yamaoka S, Kozaki T, Misawa T, Takahama M,
Satoh T, Takeuchi O, Yamamoto N, Matsuura Y, Saitoh T, Akira S. Zinc-finger
antiviral protein mediates retinoic acid inducible gene I-like receptorindependent antiviral response to murine leukemia virus. Proc. Natl.
Acad. Sci. USA. 110:12379-84, 2013.

9

Immunoglycobiology
The main topic in our laboratory has been biological and medical significance of glycosylphosphatidylinositol-anchored proteins (GPI-APs).
At least 150 different cell surface human proteins including many withimmunological functions are GPI-APs. We aim to understand molecularmechanisms of biogenesis of GPI-APs, their biological functions andmolecular pathogenesis of acquired and inherited GPI deficiencies.

1. Molecular pathogenesis of GPI-anchor deficiencies.

Taroh Kinoshita, PhD

Hyperphosphatasia mental retardation syndrome (HPMR), also Mabry
syndrome, is a recently-identified inherited GPI-anchor deficiency
caused by mutations in PIGV or PIGO, which are involved in biosynthesis
of GPI-anchor in the endoplasmic reticulum (ER). We, in collaboration
with Dr. Krawitz and his colleagues in Germany, and with Dr. Hansen in
Denmark and Dr. Abou Jamra in Germany, reported that PGAP2, whichis involved in fatty acid remodeling of GPI-APs in the Golgi apparatus,
is the third gene responsible for HPMRS (Hansen L et al, Am. J. Hum.
Genet., 92:575, 2013; Krawitz P et al, Am. J. Hum. Genet., 92:584, 2013).
We also found the first individual with inherited GPI-deficiency caused
by mutations in PIGW, involved in inositol-acylation. The individual
had hyperphosphatasia, intellectual disability and intractable seizures,
fulfilling diagnosis of HPMRS.
In collaboration with groups in Germany and England, we studiedm u t at i o n s i n P G A P 3 g e n e fo u n d i n s e ve ra l i n d i v i d u a l s w i t h
hyperphosphatasia and intellectual disability. All mutations caused
partial loss of function of PGAP3 due to mislocalization of the mutant
protein, nonsense-mediated mRNA decay, or decreased enzymatic
activity. These mutations lead to inefficient fatty acid remodeling of GPIAPs in the Golgi and result in inefficient raft-association and releaseof GPI-APs. This work revealed the importance of maturation of thelipid part of GPI-APs in neuronal development and function (Howard
MF et al, Am. J. Hum. Genet., 94:278, 2014). These results establishedthat molecular mechanisms of hyperphosphatasia (release of alkaline
phosphatase) in PIGV/PIGO/PIGW-HPMR, PGAP2-HPMR and PGAP3HPMR are different (Figure1).

2. Roles of p24γ2(TMED5)in the ER-to-Golgi transport of GPI-APs
GPI-APs are transported from the ER to the Golgi via COPII
coated transport vesicles. For efficient packaging of GPI-APs
into the COPII coated vesicles in the ER exit sites, a cargo receptor that captures and recruits GPI-APs is required. We previously reported that a complex consisting of p24α2/TMED9,p24β1/TMED2, p24γ2/TMED5 and p24δ1/TMED10, acts as a cargo receptor of GPI-APs. We recently demonstrated that p24γ2/
TMED5 is the subunit of this receptor that recognizes GPI-APs
and that the α-helical domain but not GOLD domain of p24γ2/
TMED5 is required for the recognition of GPI-APs. It is likelythat the juxta-membrane region within the α-helical domain
contains a binding site for the glycan part of GPI.
One of the goals of our research is to identify all the factors
specifically involved in transport of GPI-APs from the ER to
the cell surface membrane microdomains, and to understandfunctions of these factors. This will give us a complete pictureof how GPI-APs are synthesized and expressed in functionally
relevant subcellular locations, leading to full understandingroles of GPI-APs in various cells.

Figure1. Different mechanisms of hyperphosphatasia in PIGV/PIGO/PIGW-, PGAP2- and PGAP3-deficiencies.

Recent Publications
 Fujita M, Maeda Y, Ra M, Yamaguchi Y, Taguchi R, Kinoshita T. GPI-glycan
remodeling by PGAP5 regulates transport of GPI-anchored proteins fromthe ER to the Golgi. Cell 139:352-65, 2009.

Professor
Associate Professor

Taroh Kinoshita
Yusuke Maeda
Yoshiko Murakami

Assistant Professor

Morihisa Fujita
Noriyuki Kanzawa

10

Postdoctoral Fellow

1

Research Assistant

2

Visiting Scientist

1

Support Staff

3

 Fujita M, Watanabe R, Jaensch N, Romanova-Michaelides M, Satoh T, Kato
M, Riezman H, Yamaguchi Y, Maeda Y, Kinoshita T. Sorting of GPI-anchoredproteins into ER-exit sites by p24 proteins is dependent on remodeled GPI.
J. Cell Biol. 194:61-75, 2011.

 Kanzawa N, Shimozawa N, Wanders RJA, Ikeda K, Murakami Y, Waterham
HR, Mukai S, Fujita M, Maeda Y, Taguchi R, Fujiki Y, Kinoshita T. Defective
lipid remodeling of GPI anchors in peroxisomal disorders, Zellweger
syndrome, and rhizomelic chondrodysplasia punctata. J. Lipid Res. 53:65363, 2012.
 Wang Y, Murakami Y, Yasui T, Wakana S, Kikutani H, Kinoshita T, Maeda
Y. Significance of GPI-anchored protein enrichment in lipid rafts for thecontrol of autoimmunity. J. Biol. Chem. 288:25490-99, 2013.

 Murakami Y, Kanzawa N, Saito K, Krawitz PM, Mundlos S, Robinson PN,
Karadimitris A, Maeda Y, Kinoshita T. Mechanism for release of alkaline
phosphatase caused by glycosylphosphatidylinositol deficiency inpatients with hyperphosphatasia-mental retardation syndrome. J. Biol.
Chem. 287:6318-25, 2012.

11

Immunopathology
Our proposed study is the regulation of immune cell motility and migratory behavior in vivo by soluble and membrane-bound ‘immune
guidance molecules’ such as semaphorins and their receptors (Figure 1).
Semaphorins were originally identified as axon-guidance molecules thatfunction during neuronal development. However, cumulative evidence
indicates that semaphorins also participate in immune responses, both
physiological and pathological, and they are now considered to be potential diagnostic and/or therapeutic targets for a range of diseases. The
primary receptors for semaphorins are neuropilins and plexins, whichhave cell type-specific patterns of expression and are involved in multiple signalling responses. Beyond such basic implications, we are trying
to apply the findings from this proposed study into other research fields
and diagnosis/therapy for human diseases such as autoimmunity, allergy,
immune deficiency, cancer/metastasis, and neurodegenerative diseases.

2. Plexin-A1 is involved in Toll-like receptor‑mediated microglial activation.
Semaphorins, which are identified as repulsive axon guidance
molecules, play crucial roles in maintaining brain homeostasis
by regulating microglial activity. To investigate the in vivo roleof Plexin-A1 in lipopolysaccharide (LPS)-induced injury in thebrain, we examined the neuroinflammatory changes induced
by LPS administration in wild-type (WT) and Plexin-A1-deficient(-/-) mice. WT mice showed a significantly higher expression of
COX-2, iNOS, IL-1β and TNF-α in the hippocampus, and a significantly greater ventricular enlargement and intracerebral infiltra-tion of mononuclear cells. In contrast, Plexin-A1-/- mice administered LPS did not show a significantly increased expression of
COX-2, iNOS, IL-1β or TNF-α in the hippocampus. Plexin-A1-/mice administered LPS did not show significant increases inventricle size or infiltration of leukocytes into the brain, as compared with the saline-treated group. In WT, but not in the Plexin-A1-/- primary microglia treated with LPS, Sema3A induced
significantly more nitric oxide production than in the immunoglobulin G control. These results suggested the crucial role of
the Sema3A-Plexin-A1 interaction in the Toll-like receptor 4-mediated signaling of the LPS-induced activation of microglia.

1. Sema4A and multiple sclerosis.

Atsushi Kumanogoh, MD/PhD

Multiple sclerosis (MS) is a demyelinating autoimmune disease of the
CNS and a leading cause of lasting neurologic disabilities in young adults.
Although the precise mechanism remains incompletely understood,
approximately one-third of patients with multiple sclerosis (MS) respondpoorly to interferon-beta (IFN-β) therapy. Serum Sema4A is increasedin MS patients, and those who have high Sema4A do not respond to
IFN-β therapy. We therefore investigated whether recombinant Sema4A
abrogates the efficacy of IFN-β in mice with experimental autoimmune
encephalomyelitis (EAE), an animal model of MS. Administration of
Sema4A concurrently with IFN-β diminished the efficacy of IFN-β in
EAE. These effects of Sema4A were attributed to promote Th1 andTh17 differentiation and to increase adhesive activation of T cells toendothelial cells, even in the presence of IFN-β.

Figure 1.

Recent Publications

12

Professor

Atsushi Kumanogoh

Associate Professor

Toshihiko Toyofuku

Assistant Professor

Hyota Takamatsu

Research Assistant

6

Support Staff

1

 Nojima S, Toyofuku T, Kamao H, Ishigami C, Kaneko J, Okuno T, Takamatsu
H, Ito D, Kang S, Kimura T, Yoshida Y, Morimoto K, Maeda Y, Ogata A,
Ikawa M, Morii E, Aozasa K, Takagi J, Takahashi M, Kumanogoh A. A point
mutation in Semaphorin 4A associates with defective endosomal sortingand causes retinal degeneration. Nat. Commun. 4:1406. 2013.

 Takamatsu H, Takegahara N, Nakagawa Y, Tomura M, Taniguchi M, Friedel
RH, Rayburn H, Tessier-Lavigne M, Yoshida Y, Okuno T, Mizui M, Kang S,
Nojima S, Tsujimura T, Nakatsuji Y, Katayama I, Toyofuku T, Kikutani H,
Kumanogoh A. Semaphorins guide the entry of dendritic cells into thelymphatics by activating myosin II. Nat. Immunol. 11:594-600, 2010.

 Kumanogoh A, Kikutani H. 2013. Immunological functions of the neuropilins
and plexins as receptors for semaphorins. Nat. Rev. Immunol. 13: 802-14

 Nogi T, Yasui N, Mihara E, Matsunaga Y, Noda M, Yamashita N, Toyofuku
T, Uchiyama S, Goshima Y, Kumanogoh A, Takagi J. Structural basis forsemaphorin signalling through the plexin receptor. Nature 467:1123-7.
2010.

 Hayashi M, Nakashima T, Taniguchi M, Kodama T, Kumanogoh A,
Takayanagi H. Osteoprotection by Semaphorin 3A. Nature 485:69-74.
2012.

13

Immunochemistry

Hisashi Arase, MD/PhD

Paired receptors that consist of inhibitory and activating receptors are
involved in immune regulation. Inhibitory receptors play an importantrole to suppress immune response to self. However, some inhibitory
receptors are utilized by several viruses to evade immune response.
On the other hand, activating receptors are involved in host defense
by recognizing viral proteins. Therefore, paired receptors seem to have
evolved with pathogens and seem to play an important role in hostpathogen interactions (Arase et al. Science 2002; Shiratori et al. J.
Immunol. 2005). Based on these findings, we have been extensively
working on interactions between pathogens and various paired
receptors. These studies are important to understand not only hostpathogen interactions but also mechanisms of immune regulation.

mice and analyzed the function of PILRα. Development of
immune cells was normal in PILRα-deficient mice. However,PILRα-deficient mice were susceptible to LPS-induced
endotoxin shock. Further analyses revealed that infiltrationof neutrophils in liver and lung was significantly increased in
PILRα-deficient mice. When we analyzed neutrophils fromPILRα-deficient mice, we found that activation of integrin
by chemokine stimulation is augmented in PILRα-deficientneutrophils (Wang et al. Nat. Immunol. 2012). These findings
indicated that PILRα plays an important role in the regulation of
inflammation by regulating integrin function (Figure 1).

A) Interaction between PILR and herpes simplex virus (HSV)a) Identification of HSV receptor

D) Misfolded proteins complexed with MHC class II
molecules are targets for autoimmune diseases.

PILR is one of paired receptors that are mainly expressed on various
immune cells. PILR consists of inhibitory PILRα and activating PILRβ. We havepreviously found that both PILRα and PILRβ recognize CD99 as a host ligand(Shiratori et al. J. Exp. Med. 2004). In addition, we have identified PANP as a
new ligand for PILR (Kogure et al. Biochem. Biophys. Res. Commun. 2011).
Interestingly, specific O-glycan structures on CD99 were found to be required

MHC class II allelic polymorphisms are associated withsusceptibility to many autoimmune diseases. However,it has remained unclear how MHC class II molecules are
involved in autoimmune disease susceptibility. We foundthat cellular misfolded autoantigens are rescued fromprotein degradation by MHC class II molecules (Jiang et al.
Int. Immunol. 2013). Furthermore, we found that misfolded
proteins complexed with MHC class II molecules can becometargets for autoantibodies in autoimmune disease patients (Jinet al. Proc. Natl. Acad. Sci. USA. 2014). Misfolded proteins,which complexed with MHC class II molecules of diseasesusceptible alleles but not disease-resistant MHC class II alleles,
were recognized by autoantibodies. Autoantibody binding
to misfolded proteins transported to the cell surface by MHC
class II molecules was strongly correlated with susceptibility
to autoimmune disease. This suggested that misfolded
proteins complexed with MHC class II molecules are natural
autoantigens for autoantibodies. Indeed, expression of MHC
class II molecules is induced by inflammation during infections,and most autoimmune diseased tissues aberrantly express
MHC class II molecules. Therefore, misfolded proteins, which
normally would not be exposed to the immune system,
can be targets for autoantibodies when they avoid protein
degradation (Figure 2).

for the association with PILR (Wang et al. J. Immunol. 2008).
Because PILR is one of paired receptors, we addressed whether PILR
interacts with certain pathogens. We found that PILRα associates withglycoprotein B (gB), an envelope protein of herpes simplex virus-1 (HSV1), and the interaction between PILRα and gB is involved in membranefusion during HSV-1 infection. This suggested that interaction between
PILRα and gB plays and important role in HSV-1 infection. We showed that
immune inhibitory receptors can be exploited by viruses to invade hostcells for the first time (Satoh et al. Cell 2008; Wang et al. J. Virol. 2009).
We further analyzed host cell molecules that associate with HSV-1 gB andfound that non-muscle myosin heavy chain (NMHC-IIA) associates with gB
and is involved in HSV-1 infection (Arii et al. Nature 2010).

B) Role of Siglec in varicella zoster virus (VZV) infection

Professor

Hisashi Arase

Assistant Professor

Tadahiro Suenaga
Masako Kohyama
Kouyuki Hirayasu

Postdoctoral Fellow

1

Research Assistant2

Support Staff

3

VZV belongs to α-herpes virus similar to HSV, although cellular receptor
that mediates membrane fusion during infection was unclear. We foundthat Siglec-4 (MAG, myelin associated glycoprotein), one of paired
receptors, associates with VZV gB. Furthermore, Siglec-4 mediated
VZV infection as well as membrane fusion. Interestingly, Siglec-4 also
associated HSV gB and mediated HSV infection. Because Siglec-4 is
specifically expressed in neural tissues, Siglec-4 seemed to be involvedin neurotropic characteristics of HSV and VZV (Suenaga et al. Proc. Natl.
Acad. Sci. USA. 2010).

C) PILRα plays an important role in neutrophil infiltration
The role of PILRα in host immune response has remained unclear,although PILRα is involved in HSV-1 infection. In order to elucidate thefunction of PILRα in immune response, we generated PILRα-knockout

14

Figure 1. PILRα regulates neutrophil recruitment in inflammatory responses
via modulating integrin activation. We found that PILRα, an inhibitory
receptor containing an ITIM, negatively regulated neutrophil infiltration during
inflammation. PILRα expressed on neutrophils constitutively associated in
cis with its ligands, resulting in clustering of PILRα during stimulation with achemoattractant. Clustering of PILRα enhanced ITIM-mediated signaling, thus
modulating β2 integrin inside-out activation. These data demonstrate thatneutrophil recruitment in inflammatory responses is regulated by PILRα via
modulation of integrin activation (Wang et al. Nat. Immunol. 2013).

Figure 2. Misfolded proteins transported to the cell surface by MHC class
II molecules are targets for autoantibodies. Cellular misfolded proteins aretransported to the cell surface by MHC class II molecules without processing to
peptides when misfolded proteins are associated with MHC class II molecules
at ER (Jiang et al. Int. Immunol. 2013). Furthermore, Misfolded proteinscomplexed with MHC class II molecules of disease-susceptible alleles are
specifically recognized by autoantibodies. Of note, autoantibody binding to
misfolded proteins transported to the cell surface by MHC class II molecules
was strongly correlated with susceptibility to autoimmune disease. Thissuggested that misfolded proteins complexed with MHC class II molecules arenatural autoantigens for autoantibodies, which affects autoimmune diseasesusceptibility. (Jin et al. Proc. Natl. Acad. Sci. USA. 2014).

Recent Publications
 Jin H, Arase N, Hirayasu K, Kohyama M, Suenaga T, Saito F, Tanimura
K, Matsuoka S, Ebina K, Shi K, Toyama-Sorimachi N, Yasuda S, Horita T,
Hiwa R, Takasugi K, Ohmura K, Yoshikawa H, Saito T, Atsumi T, Sasazuki T,
Katayama I, Lanier LL, Arase H. Autoantibodies to IgG/HLA-DR complexes
are associated with rheumatoid arthritis susceptibility. Proc. Natl. Acad.
Sci. USA. 111: 3787-92, 2014.
 Jiang Y, Arase N, Kohyama M, Hirayasu K, Suenaga T, Jin H, Matsumoto M,
Shida K, Lanier LL, Saito T, Arase H. Transport of misfolded endoplasmic
reticulum proteins to the cell surface by MHC class II molecules. Int.
Immunol. 25:235-46, 2013.

 Tanaka Y, Suenaga T, Matsumoto M, Seya T, Arase H. Herpesvirus 6
Glycoproteins B (gB), gH, gL and gQ are Necessary and Sufficient for Cellto-Cell Fusion. J. Virol. 87:10900-03, 2013.
 Wang J, Shiratori I, Uehori J, Ikawa M, Arase H. Neutrophil infiltration
during inflammation is regulated by PILRα via modulation of integrin
activation. Nat. Immunol. 14:34-40, 2012.
 Suenaga T, Satoh T, Somboonthum P, Kawaguchi Y, Mori Y, Arase H.
Myelin-associated glycoprotein mediates membrane fusion and entry ofneurotropic herpesviruses. Proc. Natl. Acad. Sci. USA. 107:866-71, 2010.

15

Immune Regulation
A Summary of our Recent Research.
1. Arid5a controls IL-6 mRNA stability, which contributes toelevation of IL-6 level in vivo
Post-transcriptional regulation of IL-6 has been largely uncharacterized,with the exception of the RNase Regnase-1, which prevents autoimmunity
by destabilizing IL-6 mRNA. Here, we identified a novel RNA binding
protein, AT-rich interactive domain 5a (Arid5a), which stabilizes IL-6 but
not TNF-α mRNA through binding to the 3’ untranslated region (UTR) of
IL-6 mRNA. Arid5a was enhanced in macrophages in response to LPS, IL1β and IL-6. Arid5a deficiency inhibited elevation of IL-6 serum level in
LPS-treated mice, and suppressed IL-6 levels and the development of TH17
cells in experimental autoimmune encephalomyelitis (EAE). Importantly,
Arid5a inhibited the destabilizing effect of Regnase-1 on IL-6 mRNA.
These results indicate that Arid5a plays an important role in promotion of
inflammatory processes and autoimmune diseases.

Tadamitsu Kishimoto, MD/PhD

2. P38 MAP kinase signaling pathway regulates stabilizationof Arid5a mRNA.
In the current study, we observed that through TLR4 signaling, Arid5amRNA is induced rapidly and degrades quickly in macrophages, B cells
and dendritic cells. Interestingly, we found that through TLR4 signaling,chemical inhibitors of P38 MAP kinase, enhanced the degradation of
Arid5a mRNA levels in macrophages, B cells and dendritic cells. Thus,
we identified P38 MAP kinase signaling as a key pathway for regulating
Arid5a mRNA stability. Our findings thus indicate that p38 MAP kinasesignaling is linked to the induction of Th17 cells and autoimmune disease
through regulating Arid5a mRNA stability and its function in posttranscriptional control of IL-6 level in vivo (research in progress).

3. Aryl hydrocarbon receptor-mediated induction of miR132/212 cluster enhances TH17 cell differentiation

Professor

Tadamitsu Kishimoto

Assistant Professor

Barry Ripley

Postdoctoral Fellow

7

Support Staff

1

Aryl hydrocarbon receptor (AHR) has critical roles in autoimmune
diseases such as multiple sclerosis (MS) by controlling Interleukin 17 (IL17)-producing T helper cells (TH17 cells) and Regulatory T cells (Treg cells).
Although various transcription factors and cytokines have been
identified as key participants in TH17 generation, the role of microRNAis poorly understood. We found that miR-132/212 cluster is induced by
AHR activation under TH17-inducing, but not Treg cell-inducing conditions.
miR-132/212 cluster deficiency abrogated enhancement of TH17 celldifferentiation by AHR activation. We identified Bcl-6, a negative regulatorof TH17 cell differentiation, as a target of miR-132/212 cluster.

4. Type-I interferon controls its own production in immune
homeostasis by inducing PPAR-γ expression and an inhibitory PPAR-γ/IRF7 complex.
Type-I interferon is important for anti-viral immunity, but its overproduction is linked to the development of autoimmunity. Type-I
interferon production requires the transcription factor IRF7. How type-I
interferon signals to attenuate its own production in immune homeostasis
is not known. Here we show that type-I interferon induces expressionof PPAR-γ, which forms an inhibitory interaction with IRF7, attenuating

16

type-I interferon production via the virus-activated (MyD88independent) pathways in fibroblasts and TLR-activated (MyD88dependent) pathways in pDCs, and type-I IFN-dependent
responses in autoimmunity (manuscript under revision).

5. AHR negatively regulates type 1 interferon
production and the development of murine lupus.
In addition, we found that production of type I interferon andexpression of interferon stimulation genes in aryl hydrocarbon
receptor knockout mice are higher than in wild-type mice when
we induced lupus-like disease by pristane treatment. Our results
suggest that aryl hydrocarbon receptor is a critical negative
regulator of TLR-mediated type I interferon production andinhibits type I interferon signaling in murine lupus. Now, weare evaluating protective effects of aryl hydrocarbon receptor
agonists in murine lupus (research in progress).

6. Molecular Mechanisms of Thalidomide AntiInflammatory Effects.
Thalidomide is known to inhibit TLR4-mediated inflammatory
cytokine production (including IL-6 and TNF-alpha). We
investigated the mechanism underlying these properties bystudying the recently identified Thalidomide binding protein,
Cereblon. We demonstrated that Thalidomide derivatives
potently inhibits the TLR4-TRIF pathway (which normally actsin synergy with the MyD88-dependent pathway for cytokine
production), involving inhibition of IRF3 transcriptional activity
and production of interferon-beta. We have also demonstrated
that under homeostatic conditions cereblon exists in complex
with Rabex-5, an established regulator of intracellular transport.
Intriguingly, Treatment with Thalidomide derivatives was foundto disrupt this complex. Rabex-5 was also found to be critical for
TLR induced signal transduction. Thus, we reason that disruptionof the Cereblon-Rabex-5 complex underlies Thalidomide’s antiinflammatory properties. (manuscript submitted).

7. Therapeutic targeting of the interleukin-6 receptor.
Our research is engaged in clinical studies on the effectivenessof anti-IL6R antibody (Tocilizumab) in autoimmune diseases.
(i) Tocilizumab can inhibit bone resorption and joint
destruction in chronic rheumatoid arthritis (RA) patients by alarge-scale randomized control trial. This effect is due to theinhibitory effect of IL-6 signal blockade on the expressionof Rank-ligand and differentiation into osteoclasts ofmononuclear cells.
(ii) A randomized placebo-controlled phase III trial confirmedthat Tocilizumab is effective and safe in patients withsystemic-onset juvenile idiopathic arthritis (JIA). The USA
and EU approved the use of Tocilizumab for the treatment
of JIA. In December 2012, large-scale clinical trials for JIA
in Europe and the USA confirmed efficacy and safety ofTocilizumab.
Other autoimmune inflammatory diseases have been treatedwith Tocilizumab, including refractory relapsing polychondritis,
AA amyloidosis, reactive arthritis, polymyalgia rheumatica,
systemic sclerosis, polymyositis and acquired hemophilia A. The
results confirmed efficacy and safety of Tocilizumab in these
diseases.

Figure 1. Arid5a stabilization of IL-6 mRNA

Recent Publications
 Tanaka T, Narazaki M, Ogata A, Kishimoto T. A new era for the treatment ofinflammatory autoimmune diseases by interleukin-6 blockade strategy.
Semin. Immunol. 26:88-96, 2014.
 Nakahama T, et al. Aryl hydrocarbon receptor-mediated induction of the
microRNA-132/212 cluster promotes interleukin-17-producing T-helper
cell differentiation. Proc. Natl. Acad. Sci. USA. 110:11964-69, 2013.
 Masuda K, et al. Arid5a controls IL-6 mRNA stability, which contributes
to elevation of IL-6 level in vivo. Proc. Natl. Acad. Sci. USA. 110:9409-14,
2013.

 Nakahama T, Kimura A, Nguyen NT, Chinen I, Hanieh H, Nohara K, FujiiKuriyama Y, Kishimoto T. Aryl hydrocarbon receptor deficiency in T cells
suppresses the development of collagen-induced arthritis. Proc. Natl.
Acad. Sci. USA. 108:14222-27, 2011.
 Kimura A, Naka T, Nakahama T, Chinen I, Masuda K, Nohara K, FujiiKuriyama Y, Kishimoto T. Aryl hydrocarbon receptor in combination
with Stat1 regulates LPS-induced inflammatory responses. J. Exp. Med.
206:2027-35, 2009.

17

Mucosal Immunology
Dysregulation of innate immune responses causes immune disorders
such as inflammatory bowel diseases. Our previous study using micelacking Stat3 in their innate immune cell populations showed that
TLR-mediated activation of innate immunity, when in excess, induces
intestinal inflammation. Thus, innate immune cell activity in the intestine of normal mice is finely regulated to prevent excessive inflammatory responses. Since then, we have tried to identify unique subsets ofinnate immune myeloid cells that regulate gut immunity. As a result,we identified CD70high CD11b+ CD11c+ dendritic cells that induce Th17
cells in the intestinal lamina propria. In addition, we have identified thatCX3CR1high CD11b+ CD11c+ myeloid cells suppress T cell growth via ICAM1/VCAM-1-mediated interaction. Suppressive activity of the regulatory
subset of myeloid cells (regulatory myeloid cells: Mreg cells) is mediatedby anti-inflammatory cytokine IL-10. So far, we have characterized several
intestinal innate myeloid cell subsets that mediate the murine gut
homeostasis. However, in order to apply these basic findings to clinics
to treat inflammatory bowel diseases, we need to know whether similarsubsets of intestinal innate myeloid cells exist in the human intestine.

A role of a lymphoid tissue in the appendix.
We also analyzed the function of a lymphoid tissue in the
appendix, which is thought to be a kind of remnant tissue
in our body. Gut-associated lymphoid tissues (GALTs) such
as Peyer’s patches and isolated lymphoid follicles are known
to be responsible for generation of IgA-secreting cells. In
addition to these GALTs, there is a lymphoid cell cluster in the
appendix, which is called a cecal patch. However, the functionof the cecal patch remains unknown. We analyzed the role ofthe cecal patch in mucosal immune responses using germfree mice colonized with microbiota after appendectomy.
Appendectomized mice showed delayed accumulation of
IgA+ cells in the large intestine, but not the small intestine,
after colonization. Decreases in colonic IgA+ cells correlatedwith altered composition of fecal microbiota. IgA+ cells withphotoconverted Kaede protein in the cecal patches migrated
to the large and small intestines. In contrast, photoconverted
IgA+ cells in Peyer’s patch were preferentially recruited to thesmall intestine. Adoptive transfer of CD45.2+ cells of the cecal
patch or Peyer’s patches into CD45.1+ mice also indicated that
IgA+ cells generated in the cecal patch migrate to the large
and small intestines; IgA + cells produced in Peyer’s patches
showed preferential migration to the small intestine. IgA+ cells
in the cecal patch expressed higher levels of CCR10 than cells
in Peyer’s patches. In accordance, dendritic cells in the cecal
patch, but not Peyer’s patch, prominently induced CCR10 on
cocultured B cells. Thus, we revealed that the cecal patch is amajor site for generation of IgA-secreting cells that migrate to
the large intestine.

Figure 1. CD163low myeloid cells induced Th17 cells and its
activity was enhanced in Crohn’s diseases

Figure 2. Cecal patch generates IgA-secreting cells migrating
to the small and large intestines.

The human intestinal myeloid cell subset inducing Th17 cells.
Kiyoshi Takeda, MD/PhD

We then tried to identify a human counterpart of myeloid cell subsetthat induces Th17 cell development in the intestinal lamina propria. We
found that a CD14+ CD163low cell subset in HLA-DRhigh Lin- cells, which
was selectively present in the human intestinal lamina propria, expressed
TLR2, TLR4, and TLR5, and produced IL-6, IL-1β and TNF-α in response to
LPS. In vitro coculture with naïve T cells revealed that these cells induced
Th17 cell differentiation. We also examined CD14 + CD163 low cells in
Crohn’s disease patients. CD14+ CD163low cells from inflamed regionsof mucosa of Crohn’s disease patients expressed high levels of IL-6, IL23p19, and TNF and strongly induced Th17 cells. Enhanced Th17-inducing
activity of CD14 + CD163 low cells was also observed in non-inflamed
intestine of a patient with Crohn’s disease. Thus, we identified a humanintestinal myeloid cell subset inducing Th17 cells and the Th17-inducing
activity of the subset increases in Crohn’s disease patients.

Recent Publications

Professor

Kiyoshi Takeda

Associate Professor

Eiji Umemoto

Assistant Professor

Hisako Kayama
Makoto Kinoshita

18

Postdoctoral Fellow

1

Research Assistant

13

Visiting Scientist

1

Support Staff

2

 Masahata K, Umemoto E, Kayama H, Kotani M, Nakamura S, Kurakawa
T, Kikuta J, Gotoh K, Motooka D, Sato S, Higuchi T, Baba Y, Kurosaki T,
Kinoshita M, Shimada Y, Kimura T, Okumura R, Takeda A, Tajima M, Yoshie
O, Fukuzawa M, Kiyono H, Fagarasan S, Iida T, Ishii M, Takeda K. Generationof colonic IgA-secreting cells in the cecal patch. Nat. Commun. 10:3704,
2014.
 Hyde JL, Gardner CL, Kimura T, White JP, Trobauh DW, Huang C, Szretter KJ,
Paessler S, Takeda K, Amarasinghe G, Klimstra WB, Diamond MS. A viral
RNA structural element alters host recognition of non-self RNA. Science
343:783-87, 2014.

 Kayama H, Ueda Y, Sawa Y, Jeon SG, Ma JS, Okumura R, Kubo A, Ishii
M, Okazaki T, Murakami M, Yamamoto M, Yagita H, Takeda K. IntestinalCX3C chemokine receptor 1high (CX3CR1high) myeloid cells prevent T celldependent colitis. Proc. Natl. Acad. Sci. USA. 109:5010-15, 2012.
 Yamamoto M, Ma JS, Mueller C, Kamiyama N, Saiga H, Kubo E, Kimura T,
Okamoto T, Okuyama M, Kayama H, Nagamune K, Takashima S, Matsuura Y,
Soldati-Farve D, Takeda K. ATF6 is a host cellular target of the Toxoplasma
gondii virulence factor ROP18. J. Exp. Med. 208:1533-46, 2011.

 Ogino T, Nishimura J, Barman S, Kayama H, Uematsu S, Haraguchi N,
Uemura M, Hata T, Takemasa I, Mizushima T, Yamamoto H, Takeda K,
Doki Y, Mori M. Increased Th17-inducing activity of CD14(+) CD163(low)myeloid cells in intestinal lamina propria of patients with Crohn's disease.
Gastroenterol. 145:1380-91, 2013.

19

Molecular Immunology
1) Molecular mechanisms in immune pathology caused by hostpathogen interaction: Epstein-Barr virus (EBV)-encoded latentmembrane proteins (LMP) 1 and 2a affect B cell survival andselection
EBV infects human B cells and particularly targets the memory B
cell population for persistent infection. Among various viral genes
expressed in the EBV-infected B cells, LMP1 and 2a constitutively
activate and mimic the CD40 and BCR signals, respectively. However,it has been largely unknown what impacts these virus-derived
molecules have an in vivo activation and differentiation of B cells. In
our laboratory, we generated transgenic mice conditionally expressing
LMP1 or LMP2a in germinal center (GC) B cells to evaluate their effects
on the humoral immune responses in vivo.

Hitoshi Kikutani, MD/PhD

2) Understanding of mechanisms in emergence of auto- andpolyreactive B cells and their physiological and pathological roles

20

Hitoshi Kikutani

Associate Professor

Teruhito Yasui

Assistant Professor

Shuhei Sakakibara

Postdoctoral Fellow

1

Support Staff

2

On the other hand, several clones of anti-nuclear antibody(ANA), which is generally used for diagnosis, were isolated
from high percentage (~5%) in plasma cells from acute
SLE patients. These ANA clones are not polyreactive andrecognize each nuclear autoantigen specifically. For further
study, we plan to analyze the immunoglobulin genes
encoding ANA utilizing next generation sequencing (NGS)
technologies to understand dynamics of autoreactive B cells
for their generation and selection in humans.

In the spleen of LMP2a Tg mice, normal GC was observed whereas
titers of antigen-specific antibodies were significantly low. Sequencing
analysis of the immunoglobulin gene of antigen-specific B cells
revealed impaired affinity maturation in the LMP2a Tg mice after
immunization (Figure 1). In addition, plasma cell differentiation was
significantly accelerated in LMP2a Tg mice. These results indicate that
EBV LMP2a reduced the threshold for selection of high affinity B cells,
which may contribute to the latent infection of EBV in memory B cells.
Unlike LMP2a, the expression of LMP1 in B cells strongly inhibited
GC formation. Interestingly, GC formation and antibody response werealso impaired in chimera mice co-transferred with LMP1 Tg and wildtype-derived bone marrow cells, suggesting that LMP1+ B cells provide
inhibitory effect on neighboring wild type cells. Thus, LMP1 may
contribute to EBV infection by suppressing host humoral responses.

Professor

Is polyreactive antibody related to autoimmune diseases inhumans? As we analyzed polyreactivity of plasma cells fromsystemic erythematosus lupus (SLE) patients and healthy
donors, both contained a similar proportion of polyreactive
clones, indicating that emergence of polyreactive clones is
not correlated with disease activity of SLE. Characteristic
differences, such as affinity to antigen of polyreactive
antibodies between SLE and healthy donors remain to beelucidated.

The circulating IgG+ memory B cells from healthy individuals containa significant proportion of polyreactive clones that are reactive to
multiple self-components. Because their immunoglobulin genes
have multiple somatic hypermutations (SHM), it is expected thatpolyreactive IgG+ memory B cells are post-GC B cells. The inducersof polyreactive B cells have not been extensively investigated. Our
analysis for mouse infection model of MHV68 (murine γ-herpesvirus68) provides strong evidence that polyreactivity emerges and isselected from the splenic IgG + GC B cells harboring somaticallymutated immunoglobulins. Therefore, MHV68 infection will provide an
ideal experimental system for analysis of polyreactive B cell generation(Figure 2).

Figure 1.
EBV LMP2A reduces the threshold for selection of high-affinity B cells.
(A) The conditional expression of LMP2a by the AID (activation-induced
cytidine deaminase) promoter-driven Cre did not alter GC size andstructure in the spleen upon immunization.
(B) Generation of antigen-binding B cells was extremely impaired in the
spleen of LMP2a Tg mice after NP-CGG/alum immunization.

Figure 2.
Polyreactive clones emerges in the splenic IgG+ germinal center B
cells during MHV68 infection.
Single-cell-based cloning of immunoglobulins followed by ELISA
revealed that ~30% of the splenic IgG+ germinal center B cells showed
polyreactivity. MHV68 infection will provide ideal experimental system
for study of polyreactive B cell generation.

Recent Publications
 Kumanogoh A, Kikutani H. Immunological functions of the neuropilin andplexin as receptors for semaphorins. Nat. Rev. Immunol. 13:802-14, 2013.
 Tada S, Yasui T, Nakatsuji Y, Okuno T, Koda T, Mochizuki H, Sakoda S,
Kikutani H. BAFF controls neural cell survival through BAFF receptor. PLoS
One. 8:e70924, 2013.

 Morihana T, Goya S, Mizui M, Yasui T, Prasad DV, Kumanogoh A, Tamura M,
Shikina T, Maeda Y, Iwamoto Y, Inohara H, Kikutani H. An inhibitory rolefor Sema4A in antigen-specific allergic asthma. J. Clin. Immunol. 33:200-9,
2013.

21

Experimental Immunology

Shimon Sakaguchi, MD/PhD

Professor

Shimon Sakaguchi

Associate Professor

Naganari Ohkura
Hiroyoshi Nishikawa
Keiji Hirota

Assistant Professor

James Badger Wing
Masahide Hamaguchi
Noriko Sakaguchi

22

Postdoctoral Fellow

6

Research Assistant4

Support Staff

8

Naturally occurring regulatory T (nTreg) cells, which specifically express
the transcription factor FoxP3, are indispensable for the maintenanceof immunological self-tolerance and homeostasis. One of the main
projects in this laboratory is to understand the molecular basis of Treg
cell development and function. In the previous year, we showed that
Treg development was achieved by the combination of two independent
processes, i.e., the expression of Foxp3 and the establishment of Tregtype CpG hypomethylation, both induced by TCR stimulation. We
have continued the study and shown that developing nTreg cells
in the thymus acquire a Treg-specific and stable hypomethylation
pattern in a limited number of genes, which encode key functional
molecules including FoxP3. This epigenetic change is acquired via TCR
stimulation, beginning prior to FoxP3 expression. The Treg-specific DNA
hypomethylated regions generally act as gene enhancers in steadystate nTreg cells, contributing to the stable expression of Treg functionassociated key genes including Ctla4, Il2ra and Ikzf4 in addition to Foxp3.
Upon TCR stimulation of mature nTreg cells, FoxP3 strongly represses
many genes including Il2, contributing to Treg suppressive activity. Thus,
the Treg-specific epigenome alteration can determine the heritable
Treg-specific gene network including Foxp3-dependent gene regulation(Ohkura et al., Immunity, 2012; Ohkura et al., Immunity, 2013; Kitagawa
et al., Front. Immunol. 2013; Morikawa et al., PNAS, 2014; Morikawa etal., Immunol. Rev., 2014). Considering physiological presence of nonsuppressive FoxP3+ T cells in the immune system and loss of FoxP3 in
Treg cells under certain immunological conditions, functional nTreg cells
can be more accurately defined as a T cell subpopulation possessing theTreg-type epigenome, rather than Foxp3+ T cells. This epigenome-based
definition of Treg cells would enable better understanding of functional
stability, plasticity, and heterogeneity of Treg cells (Figure).
This laboratory also studies human Treg cells and their roles incontrolling physiological and pathological immune responses. We havepreviously shown that human FOXP3+ T cells can be dissected into
three subpopulations including suppressive and non-suppressive ones
(Miyara et al., Immunity, 2009). The most differentiated and suppressive
population is FOXP3-high CD25-high CD45RA-low cells, called effector
Treg (eTreg) cells. To enhance immune responses by specifically depleting
eTreg cells, we have been looking for a cell surface molecule that isselectively expressed by eTreg cells. One of the candidates has turned out
to be the chemokine receptor CCR4. Indeed in vitro depletion of CCR4+
cells from peripheral blood T cells selectively reduced eTreg cells andevoked immune responses against tumor-associated antigens in healthy
individuals and also in cancer patients (Sugiyama et al., PNAS, 2013). In
vivo administration of anti-CCR4 monoclonal antibody, which is now
in use to kill CCR4+ leukemic cells in adult T-cell leukemia/lymphoma(ATLL) patients in Japan, also eliminated eTreg cells and evoked immune
responses against leukemic cells by CD4 and CD8 T cells in patients.
Based on these findings we plan to embark on a new immunotherapy ofcancer this year by depleting eTreg cells and subsequently immunizing
with cancer vaccine first in ATLL patients and then in patients with othercancers (Nishikawa and Sakaguchi, Curr Opin. Immunol., 2013).

This laboratory previously established an animal modelof autoimmune arthritis, called SKG mice, which possess a
ZAP-70 gene mutation and spontaneously develop T cellmediated autoimmune arthritis immunopathologically similar
to rheumatoid arthritis (RA) in humans. This year, we haveestablished another animal model of autoimmune disease
by introducing a variety of mutations in the ZAP-70 gene inmice. One of such ZAP-70 gene knock-in mice spontaneously
developed not only arthritis but also colitis, which are
immunopathologically similar to RA and inflammatory bowel
disease, respectively, in humans. We have investigated how the
mutation affects the structure of ZAP-70 and the binding of
ZAP-70 to TCRζ chains, and downstream signal transduction,and how it consequently alters thymic selection of autoimmune
T cells and Treg cells and their respective functions. These
spontaneous models of autoimmune disease due to TCR signal
alteration are instrumental in understanding the pathogenetic
mechanisms of autoimmune diseases in humans and devising
new ways of their prevention and treatment.

Figure.
Treg-type whole gene expression profile consists of several layers of gene regulations. TCR stimulation,
possibly together with other unknown factors, induces Treg-specific epigenome changes that lead tothe constitutive expression of several key molecules including FoxP3. These key molecules control many
Treg cell-related genes including CTLA4, CD25, etc. TCR stimulation of Treg cells in the periphery not
only triggers additional key gene expression but also activates FoxP3 to exert gene repression of many
genes including Il2. These gene activations and FoxP3-dependent gene repressions at various layers
construct the whole gene expression profile of Treg cells.

Recent Publications
 Morikawa H, Ohkura N, Vandenbon A, Itoh M, Nago-Sato S, Kawaji H,
Lassmann T, Carninci P, Hayashizaki Y, Forrest A, Standley D, Date D,
Sakaguchi S, and the FANTOM consortium. Differential roles of epigenetic
changes and Foxp3 expression in regulatory T cell-specific transcriptional
regulation. Proc. Natl. Acad. Sci. USA. 111:5289-94, 2014.
 Ohkura N, Kitagawa Y, Sakaguchi S. Development and Maintenance ofRegulatory T cells. Immunity 38:414-23, 2013.
 Sugiyama D, Nishikawa H, Maeda Y, Nishioka M, Tanemura A, Katayama
I, Ezoe S, Kanakura Y Sato E, Fukumori Y, Karbach J, Jäger E, Sakaguchi S.
Anti-CCR4 mAb selectively depletes effector-type FoxP3+CD4+ regulatory T
cells, evoking anti-tumor immune responses in humans. Proc. Natl. Acad.
Sci. USA. 110:17945-50, 2013.

 Yamaguchi T, Kishi A, Osaki M, Morikawa H, Prieto-Martin P, Wing K, Saito
T, Sakaguchi S. Construction of self-recognizing regulatory T cells fromconventional T cells by controlling CTLA-4 and IL-2 expression. Proc. Natl.
Acad. Sci. USA. plus. 110:E2116-25, 2013.
 Ohkura N, Hamaguchi M, Morikawa H, Sugimura K, Tanaka A, Ito Y, Osaki
M, Tanaka Y, Yamashita R, Nakano N, Huehn J, Fehling HJ, Sparwasser
T, Nakai K, Sakaguchi S. T cell receptor stimulation-induced epigenetic
changes and foxp3 expression are independent and complementary
events required for Treg cell development. Immunity 37:785-99, 2012.

23

Cell Signaling
We have analyzed the mechanism and regulation of T cell activationand differentiation by analyzing function of genes and signaling
molecules, whose expression are regulated during T cell activation anddifferentiation. Analyzing the genes altering during T cell development,
we found that Bach2 changes the expression during T cell development.
Bach2 is transcriptional repressor and has been thought to be specific for
B cells, but we found it is expressed equally well in T cells and increase
the expression along maturation of T cells. Bach2-deficient T cells reducenaïve T cells and enhances memory-type of T cells. The Bach2-/- naïve T
cells highly express genes related to effector memory T cells, and rapidly
produce Th2 cytokines upon T cell stimulation. Re-expression of Bach2 in
effector memory T cells restored the characteristics of gene expressionof naive T cells. These findings indicate that Bach2 suppresses effector
memory-related genes to maintain the naive T cell state and regulates
generation of effector-memory T cells (Figure 1).

Takashi Saito, PhD

Figure 1. Regulation of generation of inflammatory T cells by Bach2.
Bach2 suppresses the expression of genes required to differentiate into Th2 and memory T cells, and maintainthe naïve status of T cells. Bach2-deficient T cells express genes related to inflammatory/memory T cells, and.
re-expression of Bach2 resulted in returning to the phenotype of naïve T cells

Figure 2. Costimulation of T cells by direct sensing of nucleic
acids by T cells induces Th2 differentiation.
DNAs/RNAs are recognized by unique sensor
in T cells. The nucleic acids from dying cells
directly costimulate T cells when complexedwith antimicrobial peptide LL37 or histones.
Nucleic acid-mediated ostimulation induces IL-4
production from naïve T cells and consequently
Th2 differentiation.

To understand the integrative regulation for T cell activation by various
different signals, we have analyzed the functional contribution of innate-related signals for T cell activation and function. We have previously
shown that IRAK4, a critical kinase for innate signaling is involved in T cell
activation and function. Therefore, we analyzed particularly the functionof TLRs expressed on T cells and related signals. We found that effector
Th1 but not Th2 cells were directly activated through TLR2 for IFNg
production. Nucleic acids also induce T cell co-stimulation for cytokine
production and proliferation, which we found were independent of TLR/
RLRs and their signaling. Unlike innate cells, not only CpG but also otherDNA could induce T cell co-stimulation particularly when complexedwith LL37 or histones. Nucleic acids-mediated T cell co-stimulation is induced by a yet-unknown unique sensor, and physiologically induces IL-4
production in naïve T cells which results in the induction of Th2 differentiation (Figure 2).

Recent Publications

Professor

Takashi Saito

Postdoctoral Fellow

1

Research Assistant1

 Tsukumo S, Unno M, Muto A, Takeuchi A, Kometani K, Kurosaki T, Igarashi
K, Saito T. Bach2 maintains T cells in naïve state by suppressing effector
memory-related genes. Proc. Natl. Acad. Sci. USA. 110:10735-40, 2013.
 Yokosuka T, Takamatsu M, Kobayashi-Imanishi W, Hashimoto-Tane A,
Azuma M, Saito T. Programmed cell death-1 forms negative costimulatory
microclusters that directly inhibit T cell receptor signals by recruitment ofphosphatase SHP2. J. Exp. Med. 209:1201-17, 2012.

 Kong KF, Yokosuka T, Canonigo-Balancio AJ, Isakov N, Saito T, Altman A. A
motif in the V3 domain of the kinase PKC-θ determines its localization in
the immunological synapse and functions in T cells via association with
CD28. Nat. Immunol. 12:1105-12, 2011.
 Yokosuka T, Kobayashi W, Takamatsu M, Sakata-Sogawa K, Zeng H,
Hashimoto-Tane A, Yagita, H, Tokunaga M, Saito T. Spatiotemporal basisof CTLA4-mediated negative regulation of T-cell activation. Immunity
33:326-39, 2010.

 Hashimoto-Tane A, Yokosuka T, Sakata-Sogawa K, Sakuma M, Ishihara C,
Tokunaga M, Saito T. Dynein-driven transport of T cell receptor microclusters
regulates immune synapse and T cell activation. Immunity 34:919-31, 2011.

24

25

Lymphocyte Differentiation
Introduction
B cells play an essential role in the regulation of immune responses.
Upon first encountering their cognate antigens, B cells exert multiple
functions including antibody production, antigen-presentation, andinduction of T cell differentiation. In addition to protective roles against
pathogens, B cells also have regulatory roles, e.g. serving as negative
regulators as autoimmunity by secreting anti-inflammatory cytokines
such as IL-10 and TGF-β. Our laboratory has now focused on deciphering
when and where B cells produce IL-10, thereby dampening T celldependent inflammation.
When B cells recognize the same antigen a second time, memory
antibody responses can be induced by T cell help. These are typicallyseen in the response to T cell-dependent antigens and are characterized
by the rapid production of high-titers of high-affinity antigen-specificantibody. Based upon our previous evidence that memory B cell
responses require T cells help, we are now characterizing how memory
T cells are generated and contribute to robust memory antibody
responses.

T cell differentiation have been well elucidated, its role andactivation mechanisms in TFH memory cells still remain obscure.
We demonstrated its importance for maintenance andactivation of TFH memory cells. Indeed, Bcl6 in memory TFH cells
is rapidly induced upon re-challenge by soluble antigen andthis response is mainly mediated through antigen-presentationby the memory B cells. Thus, memory B cells are the primary
APCs to induce the rapid differentiation of memory TFH cells
toward effectors, further accelerating memory B cells responses
during recall.

B cell role for immune suppression
Tomohiro Kurosaki, MD/PhD

Calcium signaling is thought to be critical for multiple B cell function;
however, supportive in vivo evidence has been lacking. Thus, ourdiscovery that Stim1 and Stim2 proteins are required in BCR-medicatedcalcium mobilization, has allowed us to approach this question by taking
a genetic approach. We generated mice with B cell-specific deletionsof both Stim1 and Stim2 and found that; although both molecules
are critically required for in vitro proliferation, they were dispensable
for B cell development and antibody responses in vivo. However, theablation of Stim1 and Stim2 in B cells caused defects in NFAT activation,
B cell intrinsic IL-10 production, and suppression of an EAE model of
autoimmune diseases. Furthermore, we demonstrated that plasmablasts,
but not B cells, indeed secrete IL-10, which in turn acts on dendritic cells(DC) cells, thereby dampening T cell-mediated inflammation.

Figure 1. The model of regulatory function of plasmablasts
Plasmablasts are generated through the interactions between B cells and CD4+ T cells.
The plasmablast-derived IL-10 inhibits dendritic cell functions and thereby suppresses
the generation of effector T cells such as Th1 and Th17 cells.

Interaction between memory T and B cells in humoral recall responses responses

Professor

Tomohiro Kurosaki

Associate Professor

Yoshihiro Baba
Wataru Ise

Assistant Professor

Takeshi Inoue
Masanori Matsumoto
Shinya Tanaka

Support Staff

26

4

In primary immune responses, it is widely accepted that among
several differentiated helper T cell subsets, follicular helper CD4 T cells(TFH cells) are the major subset to deliver help to B cells. As a TFH lineage
regulator, a transcription factor, Bcl6, has recently been identified; it
is highly expressed by TFH cells and is required for their development.
According to the current view, during a primary response, Bcl6
expression by T cells is induced by priming with dendritic cells andICOS is a key co-receptor molecule for induction of Bcl6. The initial
Bcl6 induction and subsequent cXCR5 expression allow CD4 T cells to
migrate toward the T-B border, where T FH cells interact with antigenspecific B cells. According to this model, cognate B cells are not required
for the induction of Bcl6, but support the expansion of T FH cells.
Although the importance of Bcl6 and its expression kinetics in naïve

Recent Publications
 Shinohara H, Behar M, Inoue K, Hiroshima M, Yasuda T, Nagashima T,
Kimura S, Sanjo H, Maeda S, Yumoto N, Sewon K, Akira S, Sako Y, Hoffmann
A, Kurosaki T, Okada-Hatakeyama M. Positive feedback within a kinasesignaling complex functions as a switch mechanism for NF-κB activation.
Science 344:760-64, 2014.
 Kometani K, Nakagawa R, Shinnakasu R, Kaji T, Rybouchkin A, Moriyama
S, Furukawa K, Koseki H, Takemori T, Kurosaki T. Repression of Bach2
predisposes IgG1 memory B cells toward plasma cell differentiation.
Immunity 39:136-47, 2013.

 Limnander A, Depeille P, Freedman TS, Liou J, Leitges M, Kurosaki T, Roose
JP, Weiss A. Stim1, PKCd and RasGRP set a threshold for pro-apoptotic Erk
signaling during B cell development. Nat. Immunol. 12:425-33, 2011.
 Matsumoto M, Fujii Y, Baba A, Hikida M, Kurosaki T, Baba Y. The calcium
sensors STIM1 and STIM2 control B cell regulatory function through IL-10
production. Immunity 34:703-14, 2011.

 Dong Z, Davidson D, Perez-Quintero LA, Kurosaki T, Swat W, Veillette A. The
Adaptor SAP Controls NK Cell Activation by Regulating the Enzymes Vav1 and SHIP-1 and by Enhancing Conjugates with Target Cells. Immunity
36:974-85, 2012.

27

Malaria Immunology
Our lab is interested in the host-pathogen interactions in the context
of Plasmodium infections, a disease known as malaria. We mainly focus
on how Plasmodium parasites invade host cells, mostly erythrocytes,
and cause inflammation in tissues due to infection-related events.
We try to learn from these host-parasite interactions and transfer our
understanding into the development of successful vaccines against
malaria as well as other infectious diseases.

Cevayir Coban, MD/PhD

We have developed several new experimental approaches to reveal
Plasmodium-host interactions. One of them is using new imaging
technologies such as ultra-high field MRI and multi-photon live
imaging microscopy. We have closely collaborated with other IFReC
laboratories such as Biofunctional Imaging Lab. (Prof. Yoshioka) and
Vaccine Science Lab. (Prof. Ken J Ishii) to visualize Plasmodium parasites
and related events in vivo. For instance, we recently evaluated murine
cerebral malaria pathogenesis, an experimental model for the deadliest
complication of Plasmodium falciparum infection called cerebral malaria(Zhao et al., Cell Host Microbe, 2014).
Severe symptoms of human cerebral malaria such as sudden onset
coma, death or neuro-disability are commonly observed in mice after
P. bergheiANKA infection. In this experimental cerebral malaria model,
multiple pathological events such as blood-brain-barrier disruption,
vascular leakage, immune cell accumulation (especially CD8 T cell
infiltration) occur in the brain. Although the brain is known to be affected
severely, the location and spatiotemporal mechanism of how this occurs
is poorly understood. We have identified in mice that the olfactory region
is a vulnerable location for vascular leakage during experimental cerebral
malaria. This discovery could only be possible by using an ultra-highfield MRI in combination with multi-photon live imaging microscopy. We
have shown that the olfactory bulb is the area physically disrupted anddamaged functionally by Plasmodium parasites, followed by high fever(Figure 1A). Thus, we identified that there is an early symptom, olfaction
loss, before the onset of coma. Given that trabecular small capillaries
are a unique architectural structure for olfactory bulb, olfactory easily
undergoes parasite accumulation and cell occlusion followed by microbleeding (Figure 1B).

28

Associate Professor

Cevayir Coban

Research Assistant2

Support Staff

1

In the search for the underlying mechanism(s) of pathology ofexperimental cerebral malaria via olfactory, we found that circulating
parasites in the olfactory vessels are sensed by astrocytes aroundolfactory glomeruli at the early stage of infection, and may release
CCL21and may have a role for the recruitment of pathological CD11c+
CD8 T cells into the brain. We further evaluated this novel understanding
into a novel intervention strategy by blocking chemokine-receptor
interactions when the early symptom of experimental cerebral malaria,
olfaction loss, was evident. Given that even early detection of one
day of malarial coma could dramatically increase treatment success;this previously unnoticed, truly overlooked location and detection of
olfaction loss during malaria infection may provide early, cheap and easy
diagnosis of cerebral malaria.

Figure 1.
(A) 11.7 T MRI of mouse head infected with P. bergheiANKA parasites.
Diffusion weighted image, coronal section. White dotted circle isolfactory bulb; black dark spots are bleeding spots.

(B) Fresh micro-bleeding in olfactory bulb captured by multi-photon live
imaging microscopy.

Recent Publications
 Zhao H, Aoshi T, Kawai S, Mori Y, Konishi A, Ozkan M, Fujita Y, Haseda Y,
Shimizu M, Kohyama M, Kobiyama K, Eto K, Nabekura J, Horii T, Ishino T,
Yuda M, Hemmi H, Kaisho T, Akira S, Kinoshita M, Tohyama K, Yoshioka
Y, Ishii KJ, Coban C. Olfactory is an overlooked site for the initiation of
cerebral malaria. Cell Host & Microbe. 15:551-63, 2014.

 Zhao H, Konishi A, Fujita Y, Yagi M, Ohata K, Aoshi T, Itagaki S, Sato S, Narita
H, Abdelgelil NH, Inoue M, Culleton R, Kaneko O, Nakagawa A, Horii T,
Akira S, Ishii KJ, Coban C. Lipocalin 2 bolsters innate and adaptive immune
responses to blood-stage malaria infection by reinforcing host ironmetabolism. Cell Host & Microbe. 12:705-16, 2012.

 Kobiyama K, Aoshi T, Narita H, Kuroda E, Hayashi M, Tetsutani K, Koyama
S, Mochizuki S, Sakurai K, Katakai Y, Yasutomi Y, Saijo S, Iwakura Y, Akira
S, Coban C, Ishii KJ. Nonagonistic Dectin-1 ligand transforms CpG intoa multitask nanoparticulate TLR9 agonist. Proc. Natl. Acad. Sci. USA.
111:3086-91, 2014.

 Coban C, Igari Y, Yagi M, Reimer T, Koyama S, Aoshi T, Ohata K, Tsukui T,
Takeshita F, Sakurai K, Ikegami T, Nakagawa A, Horii T, Nuñez G, Ishii KJ,
Akira S. Immunogenicity of Whole Parasite Vaccines Against Plasmodium
falciparum Involves Malarial Hemozoin and Host TLR9. Cell Host &Microbe. 7:50-61, 2010.

 Coban C, Kobiyama K, Jounai N, Tozuka M, Ishii KJ. DNA vaccines: A simple
DNA sensing matter? Hum. Vaccin. Immunother. 9:23912600, 2013.

29

Vaccine Science
The primary goal of our laboratory is to understand the immunological mechanisms of the intra- and inter-cellular signaling pathways thatmediate the immunogenicity of successful vaccines, as well as biologicalresponses to adjuvants. Such knowledge will enable us to develop novel
concepts, modalities and next generation immuno-preventive and/or
therapeutic agents against infectious diseases, cancer and allergies aswell as other non-communicable diseases.

<Basic and translational vaccine science>

Ken Ishii, MD/PhD

Nucleic acids as an essential built-in adjuvant for successful vaccines:
Our group and others have recently clarified that most successful
vaccines, such as FLU and DNA vaccines possess DNA and/or RNA, whichappear to act as essential “built-in” adjuvants (Ishii KJ et al Nature 2008,
Koyama S et al, Science Trans. Med. 2010). In 2013, we published a bookentitled “Biological DNA Sensor: The Impact of Nucleic Acids on Diseases
and Vaccinology." (Elsevier 2013 edited by Ken J. Ishii and Jason ChoonKit.)(Figure.1), which defines the meaning of DNA sensing pathways anddemonstrates the importance of the innate immune responses induced bydouble stranded DNA (dsDNA) through its influencing functions in disease
pathology and immune activity of adjuvants for vaccines.

Development of the second-generation adjuvants: CpG
oligonucleotide (ODN), a synthetic TLR9 ligand, is a promising
immunotherapeutic agent; however, it has not been successful
to develop interferon (IFN)-inducing CpG ODN forming astable nano-particle without undesirable aggregation. Here,
we generated a nano size particle CpG ODN (K3) wrappedby a non-agonistic Dectin-1 ligand schzophyllan (SPG),
namely K3-SPG. K3-SPG stimulates human PBMCs to produce
a large amount of both type-I and type-II IFN. K3-SPG thus
became a potent adjuvant, especially for CTL induction to coadministered protein antigens without conjugation that is
attributed to its nature of nano-particle, rather than targeting
Dectin-1. In addition, most of K3-SPG was rapidly trapped by
MARCO+ macrophages in the draining lymph node, and then
K3-SPG activated pDCs and CD8α+ DCs to exert its adjuvant
effects. K3-SPG acting as an influenza vaccine adjuvant wasdemonstrated in vivo by both murine and non-human primate

model. Taken together, K3-SPG may be used as IFN-inducer
as well as CTL inducer for immunotherapeutic applications(Kobiyama K et al PNAS 2014) (Figure 2). This K3-SPG has been
nominated for a JST-supported grant with a pharmaceuticalcompany in Japan.
Clinical studies on seeking bio-marker(s) for safety as
well as efficacy of adjuvanted vaccines were launched in2012 (Adjuvant Data Base project supported by Ministry of
Health, Labor and Welfare). Cohort as well as retrospective
analysis of human samples obtained from volunteers of vaccine
clinical trials and patients of relevant immunological disorders
are being conducted by four groups including our lab in
IFREC and those in NIBIO. Preliminary results suggest serummiRNA may provide useful biomarkers to predict safety andimmunogenicity of adjuvanted vaccines.

Old, but newly evolving adjuvant research; As we postulated
that our immune system is substantially modulated by metabolic
intermediates of nucleic acids (Ishii KJ et al Curr. Op Immunol 2008), we
went further on identifying a key mechanism of the most commonlyused adjuvant, aluminum salt, was due to nucleic acids as well as PGE 2,
released as an alarmin (Marichal T et al Nat. Med 2011, Kuroda E et al
Immunity 2011).
Taken together, we believe this is a new area of vaccine science and we
propose that revealing the critical role in driving the responses mediatedby many current vaccines of nucleic acid-sensing mechanisms (Desmet
C and Ishii KJ Nat Rev Immunol 2012), as well as host-derived metabolites(Jounai N et al Front Cell Infect Microbiol. 2012) and particulate molecules(Kuroda E et al, Int. Rev. Immunol 2013) may demonstrate possibilities forthe design of new vaccines.

<Human immunology, clinical development of novel
adjuvants and their biomarkers>
Professor

Ken Ishii

Associate Professor

Etsushi Kuroda

Assistant Professor

Taiki Aoshi
Kouji Kobiyama
Kohei Tetsutani

30

Postdoctoral Fellow

2

Research Assistant

4

Visiting Scientist

5

Support Staff

5

A Ph-I clinical trial for novel-adjuvanted vaccine; We have beensuccessful in developing a nucleic-acid-based adjuvant; humanized CpGODN for a travelers’ malaria vaccine targeting a blood stage parasite
antigen (Tougan T et al Human Vac 2013). Preclinical studies assessing
safety and efficacy have been completed with GMP grade humanized
CpG ODN and we conducted multiple pre-clinical studies to ensure
safety and efficacy. As a result, we obtained approval from the IRB of
Osaka University Hospital and PMDA (Japanese regulatory agency) andinitiated an investigator driven GCP Phase-I clinical trial during 2013 in
Osaka University Hospital.

Figure 1. “Biological DNA Sensor: The Impactof Nucleic Acids on Diseases and Vaccinology."(Elsevier 2013 edited by Ken J. Ishii and Choon
Kit Tang)

Figure 2. Nonagonistic Dectin-1 ligand transforms CpG into a multitask nanoparticulate
TLR9 agonist. Proc Natl Acad Sci U S A. 2014 111(8):3086-91. (Direct submission)

Recent Publications
 Kobiyama K, Aoshi T, Narita H, Kuroda E, Hayashi M, Tetsutani K, Koyama
S, Mochizuki S, Sakurai K, Katakai Y, Yasutomi Y, Saijo S, Iwakura Y, Akira
S, Coban C, Ishii KJ. Nonagonistic Dectin-1 ligand transforms CpG intoa multitask nanoparticulate TLR9 agonist. Proc. Natl. Acad. Sci. USA.
111:3086-91, 2014. (Direct submission) Kuroda E, Coban C, Ishii KJ. Particulate adjuvant and innate immunity: pastachievements, present findings, and future prospects. Int. Rev. Immunol.
32:209-20, 2013.

 Kuroda E, Ishii KJ, Uematsu S, Ohata K, Coban C, Akira S, Aritake K, Urade
Y, Morimoto Y. Silica crystals and aluminum salts regulate the productionof prostaglandin in macrophages via NALP3 inflammasome-independent
mechanisms. Immunity 34:514-26, 2011.
 Koyama S, Aoshi T, Tanimoto T, Kumagai Y, Kobiyama K, Tougan T, Sakurai
K, Coban C, Horii T, Akira S, Ishii KJ. Plasmacytoid dendritic cells delineate
immunogenicity of influenza vaccine subtypes. Sci. Transl. Med. 2:25ra24,
2010.

 Desmet CJ, Ishii KJ. Nucleic acid sensing at the interface between innate
and adaptive immunity in vaccination. Nat. Rev. Immunol. 12:479-91,
2012.

31

Immune Regulation
Innate and adaptive immunity cooperate to achieve host defense andimmune homeostasis. Ag presenting cells including dendritic cells (DCs)play critical roles in linking innate and adaptive immunity. DCs respondto a variety of immune adjuvants derived from pathogens or the hostthrough the sensors such as Toll-like receptors (TLRs). Upon sensing,DCs produce proinflammatory cytokines or type I interferons (IFNs) and
upregulate a set of costimulatory molecules, thereby inducing innate
immune responses and supporting T cell activation and differentiation.
These DC functions are critical not only for protective immunity against
pathogens but also for the pathogenesis of autoimmune and inflammatory disorders.
DCs are heterogeneous and consist of various subsets, such asplasmacytoid DC (pDC) or conventional DC (cDC), which have subsetspecific functions. We aim to clarify the molecular mechanisms forregulating these DC’s functions and to develop novel immunoregulatory
maneuvers based on the clarified mechanisms.

We are now analyzing the XCR1-DTRvenus mice for various
models including immune tolerance or disease models. Basedon the specific expression of XCR1 on CD8α/103+CD11b- cDCs,
we are also generating and analyzing several knock-in micefor clarifying the novel in vivo functions of CD8α/103+CD11bcDCs.

Critical roles of an Ets family transcription factor,
Spi-B, in pDC
pDC is featured by the ability to produce large amounts oftype I IFNs in response to signaling through TLR7 or TLR9,
which can sense host- or microorganism-derived nucleic acids.
This ability plays important roles in both protective immunity
against viral infection and pathogenesis of certain autoimmune
disorders such as SLE. It is still largely unknown how the activity
is regulated. We have found that an Ets family of transcription
factor, Spi-B, expressed abundantly in pDC, is critical for pDC

function. Spi-B could transactivate the type I IFN promoters insynergy with a transcription factor, IRF-7, which is critical for
TLR7/9-induced type I IFN gene expression. The synergy wasmost prominent with IRF-7 among IRF family members. Spi-B
also associated with IRF-7 most strongly among IRF family
members. Furthermore, Spi-B-deficient pDC and mice showed
defects in TLR7/9-induced type I IFN production.
Spi-B was also highly expressed in the intestinal microfold
cells (M cells), which are gate -keeping epithelial cells
that initiate mucosal immune responses by uptaking andtranscytosing luminal antigens. In Spi-B deficient mice, mature
M cells were absent, indicating that Spi-B is a master regulator
for M cell differentiation. We have found that Spi-B-deficient
mice are defective for mucosal IgA responses. We are now
studying how Spi-B is involved in IgA responses.

In vivo roles of a DC subset, XCR1-expressing DC
Tsuneyasu Kaisho, MD/PhD

32

Professor

Tsuneyasu Kaisho

Associate Professor

Hiroaki Hemmi

Assistant Professor

Izumi Sasaki

Research Assistant2

Support Staff

1

One cDC subset, the CD8α/103+CD11b- cDC, is characterized by thehigh ability to incorporate apoptotic or dead cells and crosspresent
antigens to generate CD8 T cell responses. The cDCs are also featuredby their high ability to produce proinflammatory cytokines in response
to various TLR signals. Through these functions, cDCs contribute to antimicrobial or anti-tumor immunity. The cDCs are also considered to beimportant for maintaining immune homeostasis. However, whether
or how the cDC subset is involved in developing various inflammatory
diseases or maintaining the immune homeostasis remains unknown.
We have generated the in vivo system for tracking or ablating
CD8α/103+CD11b- cDCs. We have focused on a chemokine receptor,
XCR1, which is highly expressed in the cDC subset. The genes for a
fluorescent protein, venus, and for the fusion protein of diphtheria
toxin receptor (DTR) and venus were k nocked into the XCR1
gene locus for generating the XCR1-venus and XCR1-DTRvenus
mice, respectively. Venus expression was selectively detected in
CD8α/103+CD11b- cDCs in various lymphoid and peripheral tissues.
Eighty to ninety percent of CD8α/103+CD11b- cDCs were venuspositive and the venus-positive cells showed much higher ability to
produce proinflammatory cytokines than the venus-negative cells
among CD8α/103+CD11b- cDCs, indicating that venus-positive cells
are functionally more mature than venus-negative cells. In the XCR1DTRvenus mice, the XCR1-expressing DCs were efficiently ablated after
injection of DT. By analyzing these mice, the XCR1-expressing DC was
found to be critical for double-stranded RNA (dsRNA)-induced CD8
T cell responses (crosspresentation) of protein, cell-associated andbacteria-derived Ags (C. Yamazaki et al. 2013). Furthermore, XCR1expressing DCs were also critical for NKT cell-mediated generation oftumor-specific memory CD8 T cell responses (K. Shimizu et al, 2013).

Figure. Xcr1+ dendritic cell deletion in mice. In the XCR1-DTRvenus mice, venus expressing cells in the CD8α/103+CD11bcDCs are ablated in vivo after injection of DT. The mice are useful for clarifying the functions of XCR1-expressing DCs.

Recent Publications
 Yamazaki C, Sugiyama M, Ohta T, Hemmi H, Hamada E, Sasaki I, Fukuda
Y, Yano T, Nobuoka M, Hirashima T, Iizuka A, Sato K, Tanaka T, Hoshino K,
Kaisho T. Critical roles of a dendritic cell subset expressing a chemokine
receptor, XCR1. J. Immunol. 190:6071-82, 2013.
 Okuma A, Hoshino K, Ohba T, Fukushi S, Aiba S, Akira S, Ono M, Kaisho T,
Muta T. Enhanced Apoptosis by Disruption of the STAT3-IκB-ζ Signaling
Pathway in Epithelial Cells Induces Sjögren’s Syndrome-like Autoimmune
Disease. Immunity 38:450-60, 2013.

 Kanaya T, Hase K, Takahashi D, Fukuda S, Hoshino K, Sasaki I, Hemmi H,
Knoop KA, Kumar N, Sato M, Katsuno T, Yokosuka O, Toyooka K, Nakai K,
Sakamoto A, Kitahara Y, Jinnohara T, McSorley SJ, Kaisho T, Williams IR,
Ohno H. The Ets transcription factor Spi-B is essential for the differentiationof intestinal microfold cells. Nat. Immunol. 13:729-36, 2012.
 Hoshino K, Sasaki I, Sugiyama T, Yano T, Yamazaki C, Yasui T, Kikutani H,
Kaisho T. Cutting edge: Critical role of IkB Kinase a in TLR7/9-Induced
type I interferon production by conventional dendritic cells. J. Immunol.
184:3341-45, 2010.

 Sasaki I, Hoshino K, Sugiyama T, Yamazaki C, Yano T, Iizuka A, Hemmi H,
Tanaka T, Saito M, Sugiyama M, Fukuda Y, Ohta T, Sato K, Ainai A, Suzuki T,
Hasegawa H, Toyama-Sorimachi N, Kohara H, Nagasawa T, Kaisho T. Spi-B
is critical for plasmacytoid dendritic cell function and development. Blood
120:4733-43, 2012.

33

Immune Network
Before joining IFReC, we were working on the molecular mechanisms
for how macrophages remove apoptotic cells. We previously identified
molecules that promote the engulfment of apoptotic cells by macrophages, and proved that the failure to remove apoptotic cells can leadto the development of lupus-like autoimmune diseases. Since we joined
IFReC in November 2011, we have started to address new research projects on the mechanisms of exosome secretion and lysosomal fusion in
macrophages and also the mechanisms that regulate hemophagocytosis
by macrophages in hyper-inflammation.

Rikinari Hanayama, MD/PhD

1. Physiological functions of exosomes and their secretionmechanisms.

2. Molecular mechanisms of lysosomal fusion in macrophages.

Exosomes are secreted small membrane vesicles, composed of a lipid
bilayer with inserted transmembrane proteins, enclosing cytosolic
components derived from the exosome-producing cells. Recently,
exosomes have received much attention as messengers of intercellularcommunication networks, allowing the exchange of proteins and lipids
between the exosome-producing cells and target cells. In particular,
the findings that exosomes carry both antigenic materials and peptideMHC complexes suggested their possible roles in triggering various
immune responses, such as antigen exchange and immune cell
activation (Figure 1). Exosomes were also shown to carry mRNAs andmicroRNAs inside them, raising the possibility that exosomes transfer
genetic information between cells. However, it is not clear whether
these processes occur under physiological conditions. The only way
to conclusively demonstrate the physiological roles for exosomes
would be to specifically inhibit or increase their secretion in vivo, anddemonstrate that this affects the physiological outcomes. Therefore,
our research is currently aimed at addressing three basic questions:

During inflammation, macrophages phagocytose many deadcells and/or bacteria into phagosomes and digest them into a
series of peptides by the fusion of phagosomes with lysosomes.
These peptides bind to MHC molecules and are transported tothe surface of macrophages by the fusion of phago-lysosomeswith cell plasma membrane. Using a similar mechanism,
undigested debris in phago-lysosomes can be released frommacrophages. During these processes, lysosomal enzymes
are also secreted, causing the degradation of the surrounding
tissues. This process is called heterolysis, but its molecularmechanisms as well as its relevance to the development ofchronic inflammation have been unclear (Figure 2).

1) What are the molecular mechanisms of exosome secretion?
2) What are the physiological functions of exosomes?
3) How do exosomes travel from donor cells to the target cells in vivo?

Associate Professor

Rikinari Hanayama

Postdoctoral Fellow

3

Research Assistant1

Support Staff

1

Figure 1. Exosomes: novel signaling entities that regulate
complex immune system.

To identify the regulator of exosome secretion, we establisheda shRNA-based high throughput screening system to sort out thecells with impaired exosome secretion. Using this assay system, werecently identified several genes that regulate the exosome secretion. We generated knockout mice of these genes to investigate thephysiological functions of exosomes in immune systems. To clarify
how exosomes travel in vivo, we generated a mouse model to visualize exosomes. As CD63 is one of the most specific protein markers
for exosomes, we generated a conditional transgenic mouse line inwhich exosomes are specifically labeled with a CD63-EGFP fusion
protein in a given immune cell population. Using this transgenic mouse, we are trying to clarify the behavior of exosomes in vivo.

We have recently identified a novel protein that can be a
mediator of lysosomal fusion in macrophages. It is a type IItransmembrane protein, carrying multiple phospholipidbinding domains in the cytoplasmic region. It is highly
expressed in various types of phagocytes, particularly
inflammatory macrophages, but not in T and B lymphocytes.
We found that this protein is specifically localized to lysosomes
and the cell plasma membrane and mediates fusion between
the membrane and lysosomes (and phago-lysosomes) upon
calcium stimuli, causing the release of undigested debris, thesecretion of lysosomal enzymes and MHC-mediated antigen
presentation. Our findings will help to elucidate the molecularmechanisms of heterolysis that can be a critical process for thedevelopment of chronic inflammation.

Figure 2. Lysosomal fusion events in macrophages.

3. Molecular mechanisms of hemophagocytosis.
Macrophages eat the apoptotic cells but not live cells byrecognizing the "eat me" signal exposed only on the apoptotic
cells. This process is strictly regulated. However, under special
conditions, macrophages go crazy and start to eat the live
blood cells. It is called hemophagocytic syndrome. It is oftencaused by severe hyper-inflammation which can occur withsystemic infection, autoimmune diseases and malignancy.
It is classified as one of the cytokine storm syndromes withhigh levels of inflammatory cytokines and also macrophageactivation syndrome that is caused by overwhelming activationof T cells and macrophages. Familial diseases are caused by
the deficiency of perforin, munc13 or syntaxin which leadto the impaired killing of virus-infected cells by NK cells andCTL and eventually cause hyper-inflammation. However,
how these hyper-inflammation induce hemophagocytosis
by macrophages remains completely unknown. To clarify themolecular mechanisms of this process, we established an invitro assay system of hemophagocytosis. Using this assay,
we have recently identified a receptor on macrophages that
promotes the hemophagocytosis. We hope that our findings
will help to clarify the pathophysiology of hemophagocytic
syndrome that currently has no effective treatment.

Recent Publications
 Hanayama R. Autoimmune Diseases and the Role of MFG-E8. MFG-E8 andInflammation, 2014. in press.

 Nagata S, Hanayama R, Kawane K. Autoimmunity and the clearance ofdead cells. Cell 140:619-30, 2010.

 Toda S, Hanayama R, Nagata S. Two-step engulfment of apoptotic cells.
Mol. Cell Biol. 32:118-25, 2012.

 Hanayama R, Miyanishi M, Yamaguchi H, Suzuki J, Nagata S. Engulfmentof apoptotic cells and its physiological roles. Cell Death. (John Wiley andSons) 165-75, 2009.

 Greer PL, Hanayama R, Bloodgood BL, Mardinly AR, Lipton DM, Flavell SW,
Kim TK, Griffith EC, Waldon Z, Maehr R, Ploegh HL, Chowdfury S, Worley PF,
Steen J, Greenberg ME. The Angelman Syndrome protein Ube3A regulates
synapse development by ubiquitinating arc. Cell 140:704-16, 2010.

34

35

Immunoparasitology
The host immune system produces inflammatory cytokines in response
to infection of an intracellular protozoan pathogen Toxoplasma gondii. T.
gondii is an obligatory intracellular protozoan parasite and the causative
agent of toxoplasmosis in humans and animals. Although the infection of
T. gondii in healthy animals largely results in opportunistic and latent infection, reactivation of this pathogen in immuno-compromised individuals suffering from AIDS or being treated by chemotherapy often leads tolethal encephalitis. Eradication of T. gondii critically requires IFN-γ, whichindirectly stimulates anti-T. gondii immune responses by robust inductionof nearly 2000 genes known as IFN-γ-inducible genes. IFN-γ activates anti-parasite programs involving IFN-γ-inducible GTPases such as p47 andp65 immunity-related GTPases called IRGs and GBPs, respectively. Mice
deficient in IRGs such as LRG-47 (also known as Irgm1), IGTP (Irgm3) or
IIGP1 (Irga6), or GBPs displayed loss of resistance to T. gondii infection.

Masahiro Yamamoto, PhD

36

Professor

Masahiro Yamamoto

Assistant Professor

Miwa Sasai

Postdoctoral Fellow

1

Support Staff

1

The immunity-related GTPases have been implicated in cellularimmunity against bacteria via autophagy. Autophagy is a homeostatic
fundamental cellular process, in which cytoplasmic cargos included indouble-membrane structures called autophagosomes are transported
to lysosomes. The autophagic process involves a series of autophagy
proteins, among which Atg5 was recently linked to the IFN-γ-mediatedparasiticidal effect against T. gondii. Myeloid-specific ablation of Atg5
in mice culminated in decreased recruitment of an IRG to T. gondii in
IFN-γ-activated macrophages and led to high parasite susceptibility invivo. Furthermore, other autophagy proteins such as Atg7 and Atg16L1
have recently been shown to play a nondegradative role in the IFN-γmediated anti-viral programs against murine norovirus, prompting us to
explore the functions of autophagy proteins other than Atg5 in the IFNγ-mediated anti-T. gondii cellular innate immune responses.
Here we have characterized the role of essential autohpagy proteins,Atg7, Atg16L1, Atg9a and Atg14 in IFN-γ-mediated inhibition of T. gondii
proliferation using mouse embryonic fibroblasts (MEFs) lacking these
proteins. MEFs lacking Atg7 or Atg16L1 exhibit defects in reductionof T. gondii by IFN-γ treatment and recruitment of an IRG Irgb6 and
GBPs to the infected parasites. In sharp contrast, Atg9a- or Atg14deficient cells show normal suppression of T. gondii growth and theaccumulation of immunity-related GTPases comparable to wild-type
cells in response to IFN-γ stimulation, indicating differential participationof autophagy regulators in the mouse system. IFN-γ stimulation leads
to strong inhibition of T. gondii proliferation in human cells. However,the contribution of human immunity-related GTPases and autophagy
proteins to the inhibition remains unknown. To analyze the role of
ATG16L1 and GBPs in human cells, we have generated ATG16L1- or GBPsdeficient human cells using Cas9/CRISPR-induced genome editing.
Although IFN-γ-induced inhibition of T. gondii proliferation are normal
in human cells devoid of ATG16L1 or GBPs, IFN-γ-induced recruitment of
GBPs to the parasites is defective in ATG16L1-deficient human cells. Thus,
these results demonstrate that IFN-γ-mediated recruitment of IRGs and
GBPs to T. gondii is differentially regulated by autophagy proteins in mice,and the mechanism for the ATG16L1-dependent GBP recruitment to theparasite may be conserved in humans.

Figure. Autophagy proteins differentially participate in IFN-γ-induced anti-T. gondii cell-autonomous immunity

Recent Publications
 Ohshima J, Lee Y, Sasai M, Saitoh T, Ma JS, Kamiyama N, Matsuura Y, PannGhill S, Hayashi M, Ebisu S, Takeda K, Akira S, Yamamoto M. Role of themouse and human autophagy proteins in IFN-γ-induced cell-autonomous
responses against Toxoplasma gondii. J. Immunol. 192:3328-35, 2014.

 Yamamoto M, Ma JS, Mueller C, Kamiyama N, Saiga H, Kubo E, Kimura T,
Okamoto T, Okuyama M, Kayama H, Nagamune K, Takashima S, Matsuura Y,
Soldati-Favre D, Takeda K. ATF6β is a host cellular target of the Toxoplasma
gondii virulence factor ROP18. J. Exp. Med. 208:1533-46, 2011.

 Meunier E, Dick MS, Dreier RF, Schurmann N, Broz DK, Warming S,
Roose-Girma M, Bumann D, Kayagaki N, Takeda K, Yamamoto M, Broz P.
Caspase-11 activation requires lysis of pathogen-containing vacuoles by
IFN-induced GTPases. Nature, 2014. in press.

 Yamamoto M, Standley DM, Takashima S, Saiga H, Okuyama M, Kayama
H, Kubo E, Ito H, Takaura M, Matsuda T, Soldati-Favre D, Takeda K. A single
polymorphic amino acid on Toxoplasma gondii kinase ROP16 determines
the direct and strain-specific activation of Stat3. J. Exp. Med. 206:2747-60,
2009.

 Yamamoto M, Okuyama M, Ma JS, Kimura T, Kamiyama N, Saiga H,
Ohshima J, Sasai M, Kayama H, Okamoto T, Huang DS, Soldati-Favre D,
Horie K, Takeda J, Takeda K. A cluster of interferon-γ-inducible p65 GTPases
plays a critical role in host defense against Toxoplasma gondii. Immunity
37:302-13, 2012.

37

Single Molecule Imaging
Theoretical and experimental studies of the mechanisms
for self/non-self discrimination by Treg cells

Toshio Yanagida, PhD

CD25+CD4+ regulatory T cells (Treg) play a key role in the discriminationof self and non-self antigens. Previously we showed that T cells requireco-signaling via CD28 or IL-2 for their proliferation and that Treg areable to inhibit the co-signaling using CTLA-4high IL-2low expression andpredominant interactions to antigen-presenting cells (APC). Morerecently we used theoretical modeling and in silico simulations to assess
whether this inhibitory mechanism is sufficient to suppress responses
to self antigens. The results indicated that the dissociation of T cells
from APC is regulated by specific Treg to discriminate self and non-self
antigens in addition to inhibiting T cell-proliferation on shared APC.
In vitro experiments showed that specific Treg actually upregulate theexpression of adhesion molecules on APC with down-regulating costimulatory ligands . Interestingly the model also predicted that Treg areable to enhance the proliferation of conventional T cells (Tconv) in thepresence of excessive co-signaling, which was confirmed in vitro. Thus,
specific Treg inhibit both the dissociation and proliferation of T cells on
APC for self-specific suppression. Therefore, the dissociation of T cells
from APC might be a crucial step to induce appropriate the expansion of
T cells .

difficult to use the same animal on different days, which isnecessary when investigating disease development over time.
We therefore have developed an implantable optical windowand surgical procedure suitable for chronic in vivo imaging.
T cell transfer model mice, which were injected with T cells
harvested from Fucci (Fluorescent Ubiquitination-based Cell
Cycle Indicator) mice, were used to visualize T cell migrationand proliferation during homeostatic proliferation and antigen
stimulation. Fucci probes label individual nuclei in G1 phasenuclei red and nuclei in S/G2/M phases green. To image thesame region on different day, the vasculature was visualizedby sulfo-rhodamine B and fluorescein (Figure .2(c)). The halflife of both angiographic agents was about 10 min, whichmeans they were available every hour for imaging. The
implantable optical window system we developed is the only
system that can observe chronically immunological events inlymph node follicles and spleen cortices for more than two
weeks. We expect this system to be capable of observing any
immunological event in both regions.

(a)Distinguishing the cell state using Raman spectroscopy

Professor
Associate Professor

Toshio Yanagida
Hideaki Fujita

We used Raman spectroscopy, which can report changes in thecell state without labeling, as a non-invasive, single cell method.
Significant differences in Raman spectra were observed at the cytosoland nucleus in different cell-lines from mouse, indicating that Raman
spectra reflect differences in the cell state. Moreover, observations
before and after the induction of differentiation in neuroblastoma
and adipocytes demonstrated that Raman spectra can detect evensubtle changes in the cell state. Differences in Raman spectra were also
observed between T cells and B cells, indicating that Raman spectra
can detect subtle changes between these two cell types (Figure. 1A).
Details of the differences were made when Raman spectroscopy was
coupled with principal component analysis (PCA), as different cell
types were positioned in different regions of the PCA score plot (Figure.
1B). Thus, the present study strongly indicates that Raman spectral
morphology, in combination with PCA, can be used to establish cellular
fingerprints that distinguish and identify cellular states. We are now
trying to visualize the transition of the T cell state after stimulation.

Tomoyuki Yamaguchi
Yutaka Komai
Postdoctoral Fellow

1

Research Assistant1

Guest Professor

Takashi Jin
Tomonobu Watanabe

38

Chronic in vivo imaging in mouse organs using an implanted chamber
To enhance the fusion between imaging and immunology, moreaccessible procedures for optical microscopy are required. Two-photon
microscopy is the only modality that can unveil cell dynamics in living
animals, but it requires invasive surgical preparation which makes it(b)
Figure 1.
(A) Raman spectrum of the fingerprint region from a T-cell (blue)
and a B-cell (red).
(B) Score plot of DAPC results from the Raman spectrum.

(c)Figure 2.
(a) Spleen window in mouse.
(b) The spleen can be seen through the window. Superimposed box corresponds tothe microscopic view.
(c) Images of angiography by sulfo-rhodamine B and fluorescein on days 14 and 15,
respectively, after antigen stimulation. White bar corresponds to 100 μm.

Recent Publications
 Ichimura T, Chiu LD, Fujita K, Kawata S, Watanabe TM, Yanagida T, Fujita
H. Visualizing cell state transition using Raman spectroscopy. PLoS One
9:e84478, 2014.
 Watanabe TM, Imada K, Yoshizawa K, Nishiyama M, Kato C, Abe F,
Morikawa TJ, Kinoshita M, Fujita H, Yanagida T. Glycine insertion makes
yellow fluorescent protein sensitive to hydrostatic pressure. PLoS One
8:e73212, 2013.

 Ikezaki K, Komori T, Yanagida T. Spontaneous detachment of the leadinghead contributes to myosin VI backward steps. PLoS One 8:e58912, 2013.
 Yasuhara N, Yamagishi R, Arai Y, Mehmood R, Kimoto C, Fujita T, Touma
K, Kaneko A, Kamikawa Y, Moriyama T, Yanagida T, Kaneko H, Yoneda
Y. Importin alpha subtypes determine differential transcription factor
localization in embryonic stem cells maintenance. Dev. Cell 26:123-35,
2013.

 Watanabe TM, Fujii F, Jin T, Umemoto E, Miyasaka M, Fujita H, Yanagida T.
Four-dimensional spatial nanometry of single particles in living cells using
polarized quantum rods. Biophys. J. 105:555-64, 2013.

39

Biofunctional Imaging
Our group has developed new highly sensitive and specific in vivo
visualization techniques with magnetic resonance imaging (MRI)and magnetic resonance spectroscopy (MRS) to unveil the functionsof integrated immune system. We applied for two patents on MRI
visualization techniques this year (on the MRI contrast probe: 2013173996, on the diagnosis and treatment: 2014-066193). This MRI contrast
probe is a magnetic substance of biological origin, biodegradable, andhas a short half-life in animal bodies. The other MRI probe was also
developed to sense pH using organic polymer (Okada et al., 2013).

Yoshichika Yoshioka, PhD

MRI is widely employed for the diagnosis of immune diseases such as
multiple sclerosis (MS: human autoimmune disease of the central nervous
system). However, the size and number of the MRI lesions have had only
modest correlations with neurological impairments in MS. Furthermore,such alterations reflect relatively advanced pathogenesis and notearlier symptoms like the breakdown of the blood–brain barrier (BBB)or edema accumulation. Auto reactive T cell accumulation in the fifth
lumbar cord (L5) was recently indicated in experimental autoimmune
encephalomyelitis (EAE) mice, an MS model. We investigated the event in
EAE mouse precisely during the disease progression by using MRI at 11.7
T (Mori et al., 2014).
Figure 1 shows T 2-weighted MRI (T 2W MRI) of a lower spinal cordof the same EAE mouse during the disease progression. We founda marked increase of T 2W MRI signal intensities in L4 and L5 during
the development of disease. At the same time, the sizes of L4 and L5changed significantly. We investigated the entire spinal cord and foundno detectable alterations in its higher levels or the brain even at thepeak phase. These results show the localized occurrence of edematous
inflammation in the lower spinal cord. This inflammation coincided with
the increase of auto reactive T cell number in the region.
Diffusion-weighted MRI (DWI) shows the signal decrease in the dorsal
marginal layers of L4 and L5 during the onset phase and further decreasein L3 and L4 during the peak phase, but recovered in L5, although theclinical scores remained high. During remission, the signal approached
the pre-clinical level and returned there by late remission. These results
suggest that changes in the DWI signal might precede clinical scores and,
therefore, act as an effective marker for disease prognosis.

Professor

Yoshichika Yoshioka

Assistant Professor

Masaki Fukunaga
Yuki Mori
Fuminori Sugihara

40

Visiting Scientist

2

Support Staff

2

We found the spinal arteries (the black regions of spine in Fig. 1) alsochanged during the disease progression. Angiographic images of MRI(MRA) showed occluded and thickened vessels, particularly during thepeak phase, followed by reperfusion in the remission phase (Figs. 1 &2). Moreover, we showed that the demyelination regions of some MS
patients, where inflammation occurs, had increased lactic acid content(Fig. 3), suggesting the presence of ischemic events.
We found that the bifurcation position of spinal arteries graduallyshifted caudally with EAE development (Fig. 2). At the peak phase, asignificant dislocation of over 2 mm between the preclinical and onset

phases was observed. We hypothesize that enlargement of thelower spinal levels caused this shift by dragging the lumbar
vessels to the lower side because there were some vessels that
did not shift with the disease development.

These results suggest that inflammation-mediated alterationsin the lower lumbar cord change the homeostasis of the spinal
cord and demonstrate that ultrahigh-field MRI enables thedetection of previously invisible pathological alterations in EAE.
MRI and MRS are non-invasive and provide diverse
information in vivo. The diverse information obtained by thesetechniques will enable direct visualization of the immune
responses in order to clarify how the integrated and dynamic
immune system actually works in the body and how immune
cells behave under pathological conditions in vivo.

Figure 1. T2W MRI of spinal cord (L3–L6) in the same EAE mouse at different stages of EAE. The spinal bodies at each stage are shown inside thedotted yellow polygons. Scale bar equals 500 μm. The top of each image
shows the dorsal side.

Figure 2. MRA of the dorsal spinal artery in an EAE mouse. (A) The sagittal
plane of the vasculature in the lower lumbar cord. The region of interest is
defined around the dorsal blood vessels by the dotted line. (B) Temporal
alterations of the dorsal spinal artery in the coronal plane. The level of the
spinal cord was registered using anatomical signals of the interspinal disks.
A bifurcation in the spinal vessels is shown by the arrows. Dislocation of
the bifurcation from the baseline (blue dotted line) was seen, and thelengths of the dislocation were correlated with the severity of the clinicalscores. Scale bar equals 1 mm.

Figure 3. 1H-MR spectra (TR/TE = 3000 ms/30 ms) at 3.0 T obtained
from a healthy control (above) and a MS patient (below). Note thepronounced increases of choline and lactate in the MS patientcompared with the control subject.

Recent Publications
 Mori Y, Murakami M, Arima Y, Zhu D, Terayama Y, Komai Y, Nakatsuji Y,
Kamimura D, Yoshioka Y. Early pathological alterations of lower lumber
cords detected by ultra-high field MRI in a mouse multiple sclerosis model.
Int. Immunol. 26:93-101, 2014.

 Ma Q, Nakane Y, Mori Y, Hasegawa M, Yoshioka Y, Watanabe T, Gonda K,
Ohuchi N, Jin T. Multilayered, core/shell nanoprobes based on magnetic
ferric oxide particles and quantum dots for multimodality imaging ofbreast cancer tumors. Biomaterials 33:8486-94, 2012.

 Okada S, Mizukami S, Matsumura Y, Yoshioka Y, Kikuchi K. A nanosphericalpolymer as an MRI sensor without paramagnetic or superparamagnetic
species. Dalton Trans. 42:15864-67, 2013.

 Mori Y, Umeda M, Fukunaga M, Ogasawara K, Yoshioka Y: MR contrast
in mouse lymph nodes with subcutaneous administration of iron oxide
particles: size dependency. Magn. Reson. Med. Sci. 10:219-27, 2011.

 Satoh T, Kidoya H, Naito H, Yamamoto M, Takemura N, Nakagawa K,
Yoshioka Y, Morii E, Takakura N, Takeuchi O, Akira S: Critical role of Trib1 indifferentiation of tissue-resident M2-like macrophages. Nature 495:52428, 2013.

41

Immunology and Cell Biology

Masaru Ishii, MD/PhD

Professor

Masaru Ishii

Assistant Professor

Junichi Kikuta
Keizo Nishikawa
Szandor Simmons

42

Postdoctoral Fellow

2

Research Assistant

2

Visiting Scientist

3

Support Staff

2

The immune system is highly dynamic and a variety of cell types
are migrating throughout the body and interacting with each other at
specific sites with an appropriate time course. The mission of the lab is tounderstand the fundamental principle controlling cellular dynamics inthis vivid small world inside our body, by means of our advanced imaging
technology. The research activity of the lab is focusing on three specifictopics, such as bone dynamics, immune and inflammatory systems, andcancer invasion, all of which are highly dynamic in vivo (see the figure).
Investigations on immune systems have been so far based on the static
histological and flow cytometry analyses and plausible ‘cartoon’ helped
us to understand their dynamic nature in vivo, although the cartoons
are sometimes incorrect. Recent advances in optical imaging technology
have enabled us to shed light on the ‘real’ dynamic phenomenon
that occurs inside the body. By using intravital 2-photon microscopy
system, our lab has originally elaborated the system for visualizing thereal cellular movements in various tissues and organs in their intact
conditions.
In particular, our lab has pioneered a novel imaging technology forvisualizing inside the bones, highly mineralized and the hardest tissues inthe body, in their completely intact conditions. By using this technique,we have first elucidated the in vivo behaviors of osteoclasts, bonedestroying special macrophages resident in bones, i.e., the migrationand positioning of their precursor macrophages, their mode of boneresorbing function in vivo and the functional and physical coupling withbone-reforming osteoblasts. By utilizing this methodology, we showedthat sphingosine-1-phosphate (S1P) controls the migratory behaviorof osteoclast precursors in concert with various chemokines (Nature
2009; J Exp Med 2010). S1P is preferentially enriched in blood plasma,and S1PR1, a cognate receptor for S1P, is expressed by bone marrow
monocytoid-lineage cells, including osteoclast precursors (Nature 2009).
Further examination demonstrated that osteoclast precursor monocytes
express S1PR2, another cognate receptor for S1P, which counteracts theaction of S1PR1 and negatively regulates migration toward S1P (J Exp
Med 2010, J Immunol 2012). Intravital imaging of bone tissues dissected
the finely tuned migratory behaviors of osteoclast precursors by these
two reciprocally acting receptors, ensuring their correct positioning on
the bone surface. We also showed the substantial contribution of S1Pmediated migratory control of bon cells by S1P by generating knockout
mice deficient for endogenous S1P transporter (J Clin Invest 2012).
Moreover, we have recently demonstrated that vitamin D, which is wellknown as a bone-protecting factor, significantly blocks bone destruction
by modulating S1P-mediated migration control of osteoclast precursor
monocytes (PNAS 2013). Based on a series of studies, we proposed a newconcept in which the migration and positioning of osteoclast precursor
monocytes on the sites to be resorbed are critical points of action in theregulation of bone destruction.
By improving bone imaging techniques and using our originallydeveloped reporter mouse strain in which a vacuolar type proton pump(a3 isoform of V-type H+-ATPase) was specifically labeled with GFP, werecently succeeded in visualizing the function of fully differentiatedosteoclasts adhering to bone surfaces in vivo (J Clin Invest 2013). This
novel visualization identified two distinct mature osteoclast functional

states; i.e., bone-resorbing (R) osteoclasts firmly adhering
to bones and devouring the bone matrix by secreting acids,
and non-resorbing (N) osteoclasts relatively loosely attached
and wriggling along the bone surface. Th17 cells, a bone
destruction-prone T cell subset, express RANKL on their surface,although its functional role remains elusive. This novel imaging
system showed that RANKL-bearing Th17 could stimulate
osteoclastic bone destruction by contacting N state osteoclasts
directly to convert them to the R state, a critical mechanism
underlying bone erosion in arthritic joints.
Chronic low-grade inflammation of adipose tissue plays acrucial role for the pathophysiology of obesity. Infiltration ofseveral immune cells such as macrophages into adipose tissue
was observed in obesity, although the initial factors triggering
their migration have not been elucidated. Here, by using
intravital multiphoton imaging technique, we analyzed thedetailed time-courses of inflammatory processes in adipose

tissues under high-fat and high-sucrose (HF/HS) diet. Mobilityof macrophages was shown to be activated just 5 days after
HF/HS diet, when the distinct hypertrophy of adipocytes
and the accumulation of macrophages have not still become
prominent. Significant increase of molecule A was detected
in mature adipocyte fraction just 5 days after HF/HS diet.
Recombinant molecule A stimulated chemotactic migration
both in vitro and in vivo, as well as induced pro-inflammatory
molecules both macrophage and adipocytes, such as TNF-aand CCL2. Finally, a neutralizing antibody targeting molecule
A efficiently suppressed the HF/HS diet-induced initialinflammatory change, i.e., increased mobilization of adipose
macrophages. In conclusion, time-lapse intravital multiphoton
imaging of adipose tissues first identified the very early event
exhibiting increased mobility of macrophages, which may betriggered by increased expression of molecule A and resultant
to progression of chronic inflammation in situ.

Figure . Sheds lights on immune cell dynamics in vivo. Immune cells are high dynamic in the living body. We
are elucidating the basic principle controlling the dynamic nature of immune cells by visualizing their in situ
behaviors with advanced imaging techniques.

Recent Publications
 Kikuta J, Kawamura S, Okiji F, Shirazaki M, Sakai S, Saito H, Ishii M. S1Pmediated osteoclast precursor monocyte migration is a critical point ofcontrol in antibone-resorptive action of active vitamin D. Proc. Natl. Acad.
Sci. USA. 110:7009-13, 2013.
 Kikuta J, Wada Y, Kowada T, Wang Z, Sun-Wada G-H, Nishiyama I, Mizukami
S, Maiya N, Yasuda H, Kumanogoh A, Kikuchi K, Germain RN, Ishii M.
Dynamic visualization of RANKL and Th17-mediated osteoclast function.
J. Clin. Invest. 123:866-73, 2013.

 Ishii M, Kikuta J, Shimazu Y, Meier-Schellersheim M, Germain RN.
Chemorepulsion by blood S1P regulates osteoclast precursor mobilization
and bone remodeling in vivo. J. Exp. Med. 207:2793-98, 2010.
 Ishii M, Egen JG, Klauschen F, Meier-Schellersheim M, Saeki Y, Vacher J,
Proia RL, Germain RN. Sphingosine-1-phosphate mobilizes osteoclast
precursors and regulates bone homeostasis. Nature 458:524-28, 2009.

 Kotani M, Kikuta J, Klauschen F, Chino T, Kobayashi Y, Yasuda H, Tamai K,
Miyawaki A, Kanagawa O, Tomura M, Ishii M. Systemic circulation andbone recruitment of osteoclast precursors tracked by using fluorescentimaging techniques. J. Immunol. 190:605-12, 2013.

43

Nuclear Medicine
Nuclear medicine is a field of great potential which can evaluate theinvivo dynamic imaging of the immune cells and molecules from small
animals to humans.
The targets of nuclear medicine imaging are metabolic responses in
inflammation, cancer, and immune related disorders by using specifictracers. To achieve these aims, we are developing a new methodology of
in-vivo PET imaging, evaluation of patients receiving therapy with newmetabolic based criteria, and new imaging modalities.
In addition to these researches, we began a twin study using 18F-FDG
PET in the 2013 fiscal year and are trying to evaluate genetic influenceon glucose metabolism in various organs. From evaluation of diffusely
increased 18F-FDG uptake in thyroid gland, we guess that there are
genetic influences on human immune function.

thyroid uptake was defined as the presence of 18F-FDG uptake
in the whole thyroid gland greater than cervical background
activity. As the result, Diffuse thyroid uptake was identified inboth sides of the twins in 5 of 31 (16%) MZ pairs but none of
DZ pairs (0%), and in either side of the twins in 6 of 31 (19%)
MZ pairs and 2 of 7 (29%) DZ pairs. Among subjects withidentifiable 18F-FDG uptake (n=18), 13 subjects had euthyroid
status. Four subjects had positive, but 14 subjects had negative
anti-TPO antibodies. We found no apparent relationshipbetween diffuse thyroid uptake and anti-TPO antibody levels or
thyroid function.

Considering the higher concordance rate in MZ thanin DZ pairs, diffusely increased 18F-FDG uptake in thyroid
gland in asymptomatic subjects is considered to be largely
influenced by genetic rather than acquired factors. (In otherwords, our immune function might be influenced by not
only environmental factors but also genetic factors.) We areplanning further analysis.

The principle of twin study

Jun Hatazawa, MD/PhD

The twin study is an excellent method of estimating the extent ofgenetic and environmental influences on associations between multiple
observed variables. The twin method is popular design in behavioral
genetics and is also applicable to non-behavioral traits. Twins are divided
into two major groups; monozygotic (MZ) twins and dizygotic (DZ) twins.
The twin study is based on the assumption that MZ twins share all of
their additive genes and DZ twins share on average half of their additive
genes. In addition, we could expect MZ and DZ twin pairs to share on
the whole their growth environment before and after birth to the same
extent. So observed differences in the phenotype of interest between
MZ twins of a pair must be due to non-genetic factors because MZ twinsare genetically identical. The addition of DZ twins allows us to separate
genetic from shared environmental factors. By comparing similarities in
MZ twins with those in DZ twins, we can estimate the extent of genetic
and environmental influences. If genetic influences are strong, similarities
in MZ twins tend to be greater than those in DZ twins. However, whenenvironmental influences are strong, the similarities in MZ twins tend tobe the same as those in DZ twins, or the similarities in both MZ and DZ
twins tend to be lower.
Our twin study was performed as a part of twin research at Osaka
University. The twin registry of our research is composed of asymptomatic
aged twin pairs, mainly over 60 years.

Recent Publications
18

Genetic influence on diffusely increased F-FDG uptake inthyroid gland in asymptomatic subjects

44

Professor

Jun Hatazawa

Research Assistant1

Support Staff

318

F-FDG accumulation in normal thyroid gland is usually not visualizedin PET study. Incidence of diffuse increase in 18F-FDG uptake has beenreported to be 0.6%-6.6%. Diffuse thyroid uptake may be a normal
variant due to genetic origin, while several reports suggested that diffuse
thyroid uptake is associated with chronic thyroiditis.
We have so far evaluated thirty-eight pairs of volunteer twins (31monozygotic (MZ) and 7 dizygotic (DZ) pairs, age ranging from 42 to 84
years, mean age 67.1 years). All of them underwent whole-body 18F-FDG
PET and laboratory tests on thyroid functions and serum anti-thyroid
peroxidase (TPO) antibody, a marker of chronic inflammation. Diffuse

 Watabe T, Shimosegawa E, Kato H, Isohashi K, Ishinashi M, Hatazawa J.
CBF/CBV maps in normal volunteers studied with 15O PET: a possible
index of cerebral Perfusion pressure. Neurosci Bull., 2014. in press.
 Watabe T, Shimosegawa E, Kato H, Isohashi K, Ishinashi M, Kitagawa K,
Fujinaka T, Hatazawa J. Paradocical reduction of cerebral blood flow after
acetazolamide loading: a hemodynamic and metabolic studies with
15O-PET. Neurosci Bull., 2014. in press.

 Miyata H, Yamasaki M, Takahashi T, Murakami K, Tanaka K, Yukinori K,
Nakajima K, Takiguchi S, Morii E, Hatazawa J, Mori M, Doki Y. Determinantsof Response to Neoadjuvant Chemotherapy for Esophageal Cancer Using(18)F-fluorodeoxiglucose Positron Emission Tomography ( (18)F-FDG-PET).
Ann. Surg. Oncol. 21:575-82. 2014.
 Yamamoto S, Watabe H, Kanai Y, Watabe T, Kato K, Hatazawa J.
Development of an ultrahigh resolution Si-PM based PET system for small
animals. Phys. Med. Biol. 58:7875-88.2013.

 Yamamoto S, Watabe H, Kanai Y, Kato K, Hatazawa J. Development of ahigh-resolution YSO gamma camera system that employs 0.8-mm pixels.
Ann. Nucl. Med. 28:232-40. 2014.

45

Chemical Imaging Techniques
Fluorogenic Probes for Quick No-Wash Live-Cell Imaging of
Intracellular Proteins

Kazuya Kikuchi, PhD

Protein labeling by synthetic fluorescence probes is a powerful
technique used to investigate protein function and localization in living
cells. In this technique, objective proteins are fused with a protein(peptide) tag and are labeled by a fluorescent probe through thebinding of the probe and tag. This method has attracted attention as
an alternative to fluorescent proteins because it has the advantage ofvarious bright fluorophores that can be incorporated into probes. In
addition, the timing of the labeling can be easily controlled, allowing
for precise spatiotemporal analyses of protein movement. On theother hand, unlabeled probes remaining inside cells have the potential
to cause an increase in the background fluorescence and hinder theidentification of labeled proteins. Therefore, rigorous washing of cells is
required to remove free probes.
We developed novel fluorogenic probes for no-wash live-cell imaging
of proteins fused to PYP-tag (Figure 1). Through the design of a new
PYP-tag ligand, specific intracellular protein labeling with rapid kinetics
and fluorogenic response was accomplished. The probes crossed thecell membrane, and cytosolic and nuclear localizations of PYP-tagged
proteins without cell washing were visualized within a 6-min reaction
time. The fluorogenic response was due to the environmental effect offluorophore upon binding to PYP-tag. Furthermore, the PYP-tag-based
method was applied to the imaging of methyl-CpG-binding domain
localization. This rapid protein-labeling system combined with the small
protein tag and designed fluorogenic probes offers a powerful method
to study the localization, movement, and function of cellular proteins.

of FLAME for in vivo targeting was evaluated in tumor-bearing
mice. FLAME was modified with polyethylene glycol (PEG)
for effective delivery to tumors, because PEGylation reduces
the uptake by increasing the electrostatic interactions withproteins and small molecules. As a result, strong 19 F MRI
signals of FLAME-PEG were observed from the tumor site,whereas the 19F MRI signals of non-PEGylated FLAME-COOH
were detected only in the liver. These results demonstratethat FLAME can be a useful 19F MRI contrast agent to overcome
two major limitations of current 19F MRI probes, that is, animpractical modifiability of the surface of nanoemulsions anda low sensitivity of small molecule-based probes.

Figure 1. Principle of no-wash labeling system based on PYP-tag.

Multifunctional Core–Shell Silica Nanoparticles for Highly
Sensitive 19F MRI
19

F magnetic resonance imaging (MRI) is an especially powerful
method for in vivo imaging of particular biomolecules, cells, and target
tissues, because of negligible background signals. In 19F MRI, fluorinatedagents need to be delivered with high density per voxel to give signalsof sufficient intensity. Therefore, highly sensitive 19F MRI contrast agentsare in great demand for their practical applications. However, we have
faced the following challenges: 1) increasing the number of fluorine
atoms decreases the solubility of the molecular probes, and 2) the
restriction of the molecular mobility attenuates the 19F MRI signals.
Professor

Kazuya Kikuchi

Associate Professor

Shin Mizukami

Assistant Professor

Yuichiro Hori
Toshiyuki Kowada
Yuko Kamikawa

Support Staff

46

1

To solve these issues, we developed a novel multifunctional 19 F
MRI contrast agent, fluorine accumulated silica nanoparticle for MRI
contrast enhancement (FLAME, Figure 2). FLAME is composed of a core
micelle filled with liquid perfluorocarbon and a robust silica shell, andhas superior properties such as high sensitivity, surface modifiability,
biocompatibility, and sufficient in vivo stability. By modifying FLAME
surface with ampicillin, gene expression of BL-tag as a reporter protein in
living cells was successfully detected using 19F MRI. Finally, the potential

Figure 2. (a) Design and (b) in vivo tumor accumulation of FLAME.

Recent Publications
 Okada S, Mizukami S, Sakata T, Matsumura M, Yoshioka Y, Kikuchi
K. Ratiometric MRI Sensors Based on Core-Shell Nanoparticles for
Quantitative pH Imaging. Adv. Mater., 2014. in press.
 Matsushita H, Mizukami S, Sugihara F, Nakanishi Y, Yoshioka Y, Kikuchi K.
Multifunctional Core-shell Silica Nanoparticles for Highly Sensitive 19F
Magnetic Resonance Imaging. Angew. Chem. Int. Ed. 53:1008-11, 2014.

 Hori Y, Nakaki K, Sato M, Mizukami S, Kikuchi K. Development ofprotein-labeling probes with a redesigned fluorogenic switch based on
intramolecular association for no-wash live-cell imaging. Angew. Chem.
Int. Ed. 51:5611-14, 2012.
 Mizukami S, Watanabe S, Akimoto Y, Kikuchi K. No-wash protein labelingwith designed fluorogenic probes and application to real-time pulsechase analysis. J. Am. Chem. Soc. 134:1623-29, 2012.

 Hori Y, Norinobu T, Sato M, Arita K, Shirakawa M, Kikuchi K. Developmentof Fluorogenic Probes for Quick No-Wash Live-Cell Imaging of IntracellularProteins. J. Am. Chem. Soc. 135:12360-65, 2013.

47

Biophotonics
Raman spectroscopic imaging uses laser based imaging to determine
the molecular composition of a sample and how the distributions ofmolecules change over time. Using only the physical scattering propertiesof the light beam, and by resonance with the endogenous molecules
in a living cell or tissue sample, it is possible to apply Raman imaging
to image molecular dynamics in living cells (Okada et al 2012). Without
using labels, the technical aspects of imaging are challenging, but bycollecting images based on the entire cellular ensemble of molecules anddecoding the resulting spectra, we can see cell components at ~500 nm
resolution, and with chemical information not available by other means.
In the Biophotonics laboratory, we are particularly interested in how theapplication of such label-free techniques can reveal new informationon the immune response, either in terms of new chemical components
appearing, in new ways that cell component distributions change, or in
how cell morphology changes during the response.

Nicholas Isaac Smith, PhD

48

Associate Professor

Nicholas Isaac Smith

Postdoctoral Fellow

3

One challenge with Raman is that the imaging rate is limited by thescattering probability of the incident light, and it has been challenging
to extract more raw signal from the cell samples. We then merged two
different imaging modes, both label-free that can provide complementary
information on the cell response (Pavillon et al 2014, Pavillon et al2013, Pavillon and Smith patent submitted 2013). The additional mode
uses coherent light and both imaging systems are spectrally isolated.
This provides rapid and quantitative phase date, at speeds higher thanwhat is achievable with spontaneous Raman imaging. The phase mode
provides quantitative spatial data, with high time resolution and theRaman mode provides chemical specificity. Merging these two modes
we get a multidimensional set of data which provides a previously
unseen view of how macrophages respond to stimulation by LPS andother pathways. The phase imaging pathway gives higher sensitivity
to proteins compared to Raman, and we found that the combinationof these provides complementary imaging data. Since Raman spectra
have hundreds or thousands of wavenumber channels, we can in theory
separately map the distributions of a wide range of molecules in the cellfrom one measurement. Practically, if the sensitivity of all molecules is
not equal, having an additional mode with higher sensitivity allows us tounravel some of the mixing of molecular distributions in the cells. This isimportant to determine which molecules are involved with, or changing
during the immune response.
Using local enhancement from inert gold nanoparticles which arereadily uptaken by macrophages, the sensitivity of Raman analysis canreach down to single molecule detection levels. This represents a typeof nano-endoscopy where the trajectory and local environment can beprobed during uptake or later cellular dynamics such as active transport(Ando et al 2011). We developed an automated process for detecting,classifying, and clustering signals from macrophage particle uptake(Pavillon et al 2013). With a robust system for treating large datasets, wewere then able to look at how different types of particle surface charges
modified the immune response (Pissuwan et al 2013), and charge effects
on uptake, sequestration and local enhancement in the macrophage(Pissuwan et al 2014). With collaborators in the Dept. Applied Physics, weare also looking at multidimensional tracking of nanoparticles
to try and understand how different proteins are involved intransport (Huang et al 2014).
Following the increasing interest of mRNA in the ensembleof cellular signaling molecules, we also created a method forquantitative analysis of the contributions of RNA bases to theRaman signature and thereby produce a metric for separating
composition and structural features of the RNA (Hobro et al2013). This has particularly good sensitivity to changes in RNA
structure and can be used to study applications in Raman
analysis of RNA. We additionally used the highly sensitive
nature of spectroscopic detection of heme-based proteins,which resonate with the incoming laser field to develop amethod for early malaria diagnosis (Hobro et al 2013), and are
now looking for ways to expand those methods into otherclinical applications.

Recent Publications
 Pissuwan D, Hobro AJ, Pavillon N, Smith NI. Distribution of label freecationic polymer-coated gold nanorods in live macrophage cells reveals
formation of groups of intracellular SERS signals of probe nanoparticles.
RSC Adv. 4:5536-41, 2014.
 Hobro AJ, Konishi A, Coban C, Smith NI. Raman spectroscopic analysis
of malaria disease progression via blood and plasma samples. Analyst.
138:3927-33, 2013.

 Hobro AJ, Standley DM, Ahmad S, Smith NI. Deconstructing RNA: opticalmeasurement of composition and structure. Phys. Chem. Chem. Phys.
15:13199-208, 2013.
 Okada M, Smith NI, Palonpon AF, Endo H, Kawata S, Sodeoka M. Label-free
Raman observation of cytochrome c dynamics during apoptosis. Proc.
Natl. Acad. Sci. USA. 109:28-32, 2012.

 Pavillon N, Hobro AJ, Smith NI. Cell Optical Density and MolecularComposition Revealed by Simultaneous Multimodal Label-Free Imaging.
Biophys. J. 105:1123-32, 2013.

49

Immune Response Dynamics
Our body consists of multiple organ systems which mutually communicate to coordinate responses to external stimuli and maintain homeostasis of internal environments. Thus, to understand biological eventsoccurring in complex organ systems in our body, it is important to reveal
the interconnection among multiple organ systems rather than focus on
an isolated organ system. Keeping this notion in mind, we are investigating the communication between the nervous and immune systems. It
has long been proposed that some aspects of immune responses wereaffected by activities of the nervous system. Indeed, the autonomic nervous system was shown to modulate the pathology of immune disorders,
including rheumatoid arthritis and multiple sclerosis (Bellinger, et al.
Cell. Immunol. 252: 27, 2008). Lymphoid organs are innervated by neurons releasing a variety of neurotransmitters, and immune cells express
corresponding neurotransmitter receptors, of which stimulation affects
a broad range of immune cell activities, including proliferation and cytokine production (Tracey. Annu. Rev. Immunol. 30: 313, 2012). However, thecellular and molecular basis by which the inputs from the nervous system are converted to the outputs from the immune system is still largely
unclear.

Kazuhiro Suzuki, MD/PhD

Precise trafficking and positioning of immune cells are essential
for homeostasis of the immune system and induction of immune
responses, most part of which is orchestrated by a family of G proteincoupled receptors (GPCRs) that respond to chemoattractive molecules
represented by chemokines. As well as chemokine receptors, many of theneurotransmitter receptors are also GPCRs. A recent study showed thatdifferent types of GPCRs form heteromeric complexes on the cell surface
and cross-regulate their signaling (Fribourg, et al. Cell 147: 1011, 2011).
This observation prompted us to investigate the relationship between
neurotransmitter and chemokine receptors. We found that a group of
neurotransmitter receptors expressed in lymphocytes form complexes
with specific chemokine receptors on the cell surface. Stimulationof the neurotransmitter receptors in lymphocytes modulated the
responsiveness of the associated chemokine receptors, which had a great
impact on lymphocyte recirculation through lymphoid organs in mice.
Moreover, alteration of lymphocyte dynamics by the neurotransmittersaffected the disease course of mouse models of autoimmunity.

In this study, we identified a lymphocyte-intrinsic mechanism
by which neuronal inputs control migratory behaviors oflymphocytes and revealed a novel layer of immune regulationby the nervous system. However, it remains unclear how
neurons interact with lymphocytes and control their trafficking
in vivo. To solve this issue, we are setting up an experimental
system to visualize the interface between the nervous
and immune systems in a single frame by in vivo imaging
using two-photon microscopy. This would help capture areal picture of the communication between the two organsystems. Another direction of our study is to clarify molecularmechanisms of the crosstalk between neurotransmitter andchemokine receptors. We have identified candidate moleculesthat could be involved in the crosstalk between the two classesof receptors. Functional analysis of these molecules would
further promote our understanding of the molecular basis forneural regulation of immunity.

Figure. Control of lymphocyte dynamics by neuronal inputs. The crosstalk between the neurotransmitter andchemokine receptors regulates lymphocyte trafficking among lymphoid organs.

Recent Publications

Associate Professor

Kazuhiro Suzuki

Research Assistant1

Support Staff

3

 Gray EE, Friend S, Suzuki K, Phan, TG, Cyster JG. Subcapsular sinus
macrophage fragmentation and CD169+ bleb acquisition by closely
associated IL-17-committed innate-like lymphocytes. PLoS One 7:e38258,
2012.
 Wang X, Cho B, Suzuki K, Xu Y, Green JA, An J, Cyster JG. Follicular dendritic
cells help establish follicle identity and promote B cell retention ingerminal centers. J. Exp. Med. 208:2497-510, 2011.

 Gray EE, Suzuki K, Cyster JG. Identification of a motile IL-17-producing
gamma delta T cell population in the dermis. J. Immunol. 186:6091-95,
2011.
 Suzuki K, Grigorova I, Phan TG, Kelly LM, Cyster JG. Visualizing B cell
capture of cognate antigen from follicular dendritic cells. J. Exp. Med.
206:1485-93, 2009.

 Green JA, Suzuki K, Cho B, Willison D, Palmer D, Allen CDC, Schmidt TH, Xu
Y, Proia R, Coughlin S R, Cyster JG. The sphingosine 1-phosphate receptor
S1P2 maintains the homeostasis of germinal center B cells and promotes
niche confinement. Nat. Immunol. 12:672-80, 2011.

50

51

Information systems
We did the following studies in Information Systems.
Ultrasonic Imaging
1. Image Registration :We proposed ultrasonic image registration for multi-frequency
analysis (Figure 1). The goal of our research is the portable and real timebrain diagnosis under the thick-skull. The choice of ultrasonic frequency
is a trade-off between spatial image resolution and imaging depth. This
study shows the usability of data synthesis by employing two different
frequency ultrasounds. In the first part of this study, using Fast Fourier
Transform, we conclude that the synthesized image was produced
from two ultrasonic images of individual objects. The purpose of thesecond approach of the data synthesis is to investigate three methods ofultrasonic imaging. This approach is of particular interest for the designof further study intending to visualize any defects by ultrasonic methods.
As for the results, the synthesized image with Wavelet transform has
higher efficiency than the other synthesized images for the bone andthe sulcus. In summary, this study indicates that ultrasonic synthesizedimaging is useful to visualize the imitated brain area. This observation is
encouraging for further studies of evaluating the brain in patients.

Yutaka Hata, PhD

Professor

Yutaka Hata

Associate Professor

Syoji Kobashi
Kei Kuramoto

52

Assistant Professor

Manabu Nii

Support Staff

1

deformable surface model based on the fuzzy models to thesubject's head MR images, and estimates the subject's brain
region. To validate the proposed method, it has been applied
to 10 newborn subjects (revised ages are −1month and 1
month), and compare the segmentation result with those of

2. Ultrasonic Average Difference Imaging:
We proposed fuzzy average difference imaging for ultrasonicnondestructive testing. In our experiment, we employed a piece of windturbine blade as a specimen. The specimen has holes on the rear side as
artificial damage. We acquired ultrasonic waveforms from scanning lines
on the surface of the specimen using an ultrasonic single probe. We madecross-sectional images of the specimen by correcting each scanning line
wave data. We set scanning lines so that specimen constructions under
the lines are the same as each other. Therefore the images show the same
construction of inside the specimen, we can enhance the damage echoesby using average difference imaging. To extract the damage echoes fromthe images, we applied a damage extraction method aided by fuzzy logic
and average difference imaging. As a result, we found the line image withall damage portions, and we estimated the depth of damage surface
with high accuracy. Therefore fuzzy average difference imaging showed
effectiveness for extracting difference portions on similar images.
Magnetic Resonance image processing
Hypoxic-ischemic encephalopathy is a typical cerebral disorder ofnewborn babies. For diagnosis of neonatal cerebral disorders, themeasurement of cerebral volume and surface area is effective forquantitatively evaluating the morphological change. The measurement
requires a brain segmentation process in magnetic resonance (MR)images. However, there are few studies for newborn brain segmentation.
This study proposes an automated brain segmentation method for the
newborn brain in MR images. The automated method can improve thediagnosis of the newborn cerebral disorders with efficiency and highaccuracy. The proposed method first constructs fuzzy models using
learning dataset (Figure 2). The fuzzy models express brain features by
fuzzy membership functions. Next, the proposed method applies thethe conventional methods. Leave-one-out-cross validation
test was conducted. The mean accuracy was 93.6 ± 3.7%, themean sensitivity was 98.9±0.5, and the mean G-metrics was
4.4 ± 0.8.

Figure 1. Ultrasonic image registration

Figure 2. Growing fuzzy anatomical model of the neonatal brain.

Recent Publications
 Yagi N, Oshiro Y, Ishikawa T, Hata Y. YURAGI synthesis for ultrasonic humanbrain imaging. JACIII 17:74-82, 2013.
 Kobashi S, Kuramoto K, Wakata Y, Ando K, Ishikura R, Ishikawa T, Hirota S,
Hata Y. Newborn brain MR Image segmentation using deformable surface
model based on fuzzy knowledge models. IC-MED 5:115-24, 2013.
 Yagi N, Hata Y, Shibanuma N. Fuzzy support system for total hip
arthroplasty stem by ultrasonic intraoperative measurement. Advances inFuzzy Systems doi:10.1155/2012/201248, 2012.

 Hata Y, Kobashi S, Kuramoto K, Nakajima H. Fuzzy Biosignal Detection
Algorithm and Its Application to Health Monitoring. ACMIJ 10:133-45,
2011.
 Oe K, Miwa M, Nagamune K, Sakai Y, Lee S. Y, Niikura T, Iwakura T,
Hasegawa T, Shibanuma N, Hata Y, Kuroda R, Kurosaka M. Nondestructive
evaluation of cell numbers in bone marrow stromal cells/beta-tricalcium
phospate composites using ultrasound. Tissue Engineering, Part C:
Methods 16:347-53, 2010.

53

Systems Immunology
Our Laboratory uses computational methods to address biologicalquestions that cannot easily be elucidated by experiment. We collaborateclosely with a number of experimental groups and also develop softwaretools for general use. Some projects we are actively working on are
described below.

Structures of protein-RNA complexes
Recent studies have revealed that control of gene expression at themRNA level is essential for proper immune function. RNA-binding
proteins (RBPs) have been shown to be important for controlling thestability of mRNAs coding for a number of essential proteins (e.g.,
Uehata, 2013). In order to reveal the mechanism by which RBPs recognize
their RNA targets, we are developing in silico methods for predicting
the structures of protein-RNA complexes. A key step in our approachis predicting RNA binding sites on the molecular surface of a protein
structure of interest. To this end, we have developed a web-based
tool called aaRNA, which out-performs established methods in threepublished benchmarks (Figure 1).

Multiple sequence alignment
Multiple sequence alignment (MSA) is at the core of manybioinformatics methods, including structural modeling,evolutionary analysis and function prediction. It is well establishedthat structural information can be used to improve the accuracy
protein MSAs; however, the optimal integration of sequence andstructural information is far from trivial. Our approach to solving
this problem (Katoh, 2013), which we call MAFFTash (http://sysimm.
ifrec.osaka-u.ac.jp/MAFFTash/), compares very favorably with two
established methods (Figure 3).

Macromolecular Dynamics
We use molecular dynamics (MD) to study proteins and protein
complexes. Because we can observe the effects of ligand binding,point mutations, changes in temperature, etc. at high temporaland spatial resolution, MD is an important tool for understanding
biomolecular function.

Figure 2. EOur performance in a recent antibody modeling
contest.

Figure 1. A comparison of aaRNA with two state-of-the-art methods forpredicting protein-RNA interactions.

Figure 3. A comparison of MAFFTash and two state-of-theart multiple sequence alignment methods.

Antibody modeling and engineering

Daron M. Standley, PhD

Antibodies are members of one of the largest protein superfamilies, andare currently the fastest growing class of therapeutic agents. To facilitateengineering of customized antibodies, we have developed a pipeline
called Kotai Antibody Builder (http://kotaiab.org/). In a recent blind testof antibody structural modeling methods (AMA-II), our joint team with
Astellas Pharma and the Institute for Protein Research (Shirai, 2014)
succeeded in submitting the models of with the average lowest error, bothin terms of whole variable region and the CDR-H3 loop (Figure 2).

Structures of TLR signaling complexes
TLR signaling is mediated by adaptor molecules that share a Toll/
Interleukin-1 receptor (TIR) domain. A number of previous studies haveproposed pairwise interactions between these TIR domain-containing
proteins based on mutagenesis data; however, many of these models
remain controversial. Using a novel flexible docking procedure, weconstructed an atomic-resolution structure of the trimeric complex
formed by one TRIF and two TRAM TIR domains based on mutagenesis
and yeast two-hybrid (Y2H) data (Enokizono, 2013). The resulting
complex helps explain how the TIR domain-mediated signal propagates.

Mechanism of STIM1 calcium sensing
Associate Professor

Daron M. Standley

Associate Professor

Kazutaka Katoh

Assistant Professor

Shide Liang
Alexis Vandenbon

54

Postdoctoral Fellow

5

Research Assistant2

Support Staff

1

The protein STIM1 is a key regulator of Store-operated Ca2+
entry (SOCE). Several studies in mice have indicated that SOCE is
physiologically important in mast cells, T cells, B cells, and skeletal
muscles. Interestingly, STIM1 responds cooperatively to Ca2+ depletionin the ER, resulting in calcium influx at precise Ca2+ concentrations. We
dissected the mechanism of STIM1-mediated calcium signaling andfound that loss of calcium promotes unfolding and dimerization in acooperative manner. In vitro experiments and mathematical modeling
could recapitulate the dissociation constant, free energy, Hill coefficient,and critical Ca2+ concentration of the in vivo reaction, supporting the
proposed mechanism (Furukawa, 2014).

Recent Publications
 Furukawa Y, Teraguchi S, Ikegami T, Dagliyan O, Jin L, Hall D, Dokholyan
NV, Namba K, Akira S, Kurosaki T, Baba Y, Standley DM. Intrinsic DisorderMediates Cooperative Signal Transduction in STIM1. J. Mol. Biol. 15:208297, 2014.
 Uehata T, Iwasaki H, Vandenbon A, Matsushita K, Hernandez-Cuellar E,
Kuniyoshi K, Satoh T, Mino T, Suzuki Y, Standley DM, Tsujimura T, Rakugi H,
Isaka Y, Takeuchi O, Akira S. Malt1-induced cleavage of regnase-1 in CD4(+)
helper T cells regulates immune activation. Cell 153:1036-49, 2013.

 Katoh K, Standley DM. MAFFT multiple sequence alignment software
version 7: improvements in performance and usability. Mol. Biol. Evol.
30:772-80, 2013.
 Enokizono Y, Kumeta H, Funami K, Horiuchi M, Sarmiento J, Yamashita K,
Standley DM, Matsumoto M, Seya T, Inagaki F. Structures and interface
mapping of the TIR domain-containing adaptor molecules involved ininterferon signaling. Proc. Natl. Acad. Sci. USA. 110:19908-13, 2013.

 Shirai H, Ikeda K, Yamashita K, Tsuchiya Y, Mizuguchi K, Higo J - i, Standley
DM, Nakamura H. High-resolution structural model building of antibodies
by combination of bioinformatics, expert knowledge and molecularsimulations. Proteins doi:10.1002/prot.24591, 2014.

55

Quantitative Immunology Research
MINING BIOLOGICAL NETWORKS
The immune system is a complex dynamical network of interconnected,
hierarchically organized tissues, cells and molecules, that prevents
infection by pathogens and maintains homeostasis. Understanding
the whole dynamics of the immune system is critical for the properdevelopment of preventive and curative therapies against diseases. Thisrequires the integration of information at different levels (e.g. tissue, celland molecule), over time, in a quantitative fashion, and their abstraction
into mathematical models that enables predicting future behavior.
The Quantitative Immunology Research Unit is a team of researcherswith expertise in different scientific fields including immunology,
bioinformatics and theoretical physics. Our aim is to analyze theimmune system through the mining of biological networks, by using
three different but closely interconnected approaches; (1) quantitative
measurement of molecular dynamics, (2) integration of “big data”
from multiple sources into network models, and (3) development of amathematical framework to understand the dynamics of this massive
data. These approaches are combined in several projects that aim to gain
insight into specific problems related to the immune system. Some ofthese projects are described below to highlight specific topics.

QUANTITATIVE APPROACHES
Accurate quantification of biological responses is critical forunderstanding the dynamics of complex systems. Previously, we havedeveloped a fluorescent protein reporter system for the quantitative
monitoring of IFN-a6 (Kumagai et al. 2007). Now, we are trying to increase
the “dimension” of the observation in two ways: time and perturbation.
Time lapse imaging of type I interferon expression under microscope will
be combined with multiple fluorescent protein knock-in cells to monitor
genes induced upon antiviral responses such as IL-6, IL-10.
In spite of the importance of receptor molecule dynamics such as
dimerization and clustering with downstream molecules, this process
is still poorly understood. To address this problem, we are applying, incollaboration with RIKEN QBiC and other laboratories in IFReC, totalinternal reflection fluorescent microscopy (TIRFM) to monitor dynamics
of single immune molecules. We have succeeded in monitoring TLRs andtheir adaptors, and hope this highly quantitative technique can be used
to describe detailed dynamics, possibly leading to rational drug design.

DATA INTEGRATION

Assistant Professor

Diego Diez
Yutaro Kumagai
Shunsuke Teraguchi

56

Postdoctoral Fellow

1

Research Assistant1

The development of high-throughput technologies has brought
biology into the omics era and the need for “big data” approaches
that integrate, summarize and extract the relevant information, while
preserving the critical relation between biological components. We
make heavy use of experimental approaches to measure transcriptome,
cistrome (transcription factor binding locations) and proteome levels
over time, and integrate this information with protein-protein interaction
data, which allow us to obtain insight into signaling and gene regulatory
networks.

For example, we have combined several omics datasets togain insight into hematopoietic differentiation. One of themost important questions in immunology is how immune cells
differentiate from the original hematopoietic progenitors into
cells capable of eliciting appropriate responses. This process
is partly controlled by transcription factors (TFs) forming
complexes known as transcriptional regulatory modules (TRMs)
that regulate transcriptional programs. We have developed amethod called rTRM to identify TRMs from PPI networks (Diezet al. 2014), and applied it to publicly available transcriptional
profiles obtained for diverse immune cells (i.e. B-, T-, NKcells, dendritic cells, monocytes and macrophages) during
hematopoietic differentiation. We identified TRMs defining cell
type specificity, and predicted transcription factors with criticalroles during differentiation of hematopoietic cells.

Figure 1. The Quantitative Immunology Research Unit combines
quantitative, integrative and theoretical approaches.

MATHEMATICAL MODELINGIn many other disciplines in science, theoretical framework,
typically represented in terms of mathematical language, plays
a crucial role. However, advances in theoretical immunology
have been hampered by the lack of comprehensive accurate
measurements of biological phenomena. The quantificationof immunological responses and the integration of massive
data open the door to construct such theoretical models
in immunology. For this purpose, we are developing novel
mathematical frameworks for the description of the immune
system.
For example, in one of our collaborative researches,
we obtained quantitative data of the cooperative STIM1
dimerization during signal transduction upon activation by
decrease in Ca 2+ concentration. We found that cooperative
behavior cannot be described by the conventional biochemical
equations due to the disorder nature of the protein, and hence,a suitable theoretical framework was needed for understanding

the phenomena. We successfully developed a statisticalmechanics-inspired formulation. The model quantitatively
agreed with experimental data and enabled us to narrow
down the dimerization site of STIM1, which was reinforced bysubsequent experiments (Furukawa et al. 2014).
We have also developed a mathematical framework(Stochastic Binary Modeling, SBM) to circumvent the necessityof biochemical determination of many parameters, which
is a common obstacle for constructing a dynamical modelof cellular signaling (Teraguchi et al. 2011). SBM also allows
us to represent the stochastic and heterogeneous natureof cell populations. Currently, we are developing a system
to automatically identify the structure and parameters of
the network of regulatory pathways with the help of data
assimilation techniques. We expect that this new system will
become a useful tool for mining the immune system’s networks.

Recent Publications
 Furukawa Y, Teraguchi S, Ikegami T, Dagliyan O, Jin L, Hall D, Dokholyan
NV, Namba K, Akira S, Kurosaki T, Baba Y, Standley DM. Intrinsic DisorderMediates Cooperative Signal Transduction in STIM1. J. Mol. Biol. 15:208297, 2014.

 Patil A, Kumagai Y, Liang KC, Suzuki Y, Nakai K. Linking transcriptionalchanges over time in stimulated dendritic cells to identify gene networks
activated during the innate immune response. PLoS Comput. Biol.
9(11):e1003323, 2013.

 Diez D, Hutchins AP, Miranda-Saavedra D. Systematic identification oftranscriptional regulatory modules from protein-protein interaction
networks. Nucleic Acids Res. 42(1):e6, 2014.

 Teraguchi S, Kumagai Y, Vandenbon A, Akira S, Standley DM. Stochastic
binary modeling of cells in continuous time as an alternative tobiochemical reaction equations. Phys. Rev. E. 84(6): p. 062903, 2011.

 Hutchins AP, Diez D, Miranda-Saavedra D. Genomic and computational
approaches to dissect the mechanisms of STAT3's universal and cell typespecific functions. Jakstat. 2(4):e25097, 2013.

57

Next Generation Optical Immune-imaging
This unit started on November 1, 2013. Our aim is to understand thecomplex dynamic mechanisms of cell-cell fusion with novel imaging
technologies.
Myeloid cells such as macrophages have a pronounced potential
to fuse not only among themselves (homotypic fusion) but also withother cell types (heterotypic fusion). Homotypic cell-cell fusion leads
to formation of multinucleated osteoclasts and/or multinucleatedgiant macrophages. The role of this type of fusion is thought toenhance cellular functions. For example, osteoclasts are specialized
multinucleated giant macrophages which resorb bone. Multinucleation
allows osteoclasts to resorb larger areas, and mutant osteoclasts,
which are not able to fuse, exhibit impaired bone resorption activity.
On the other hand, some macrophages have a potential to fuse withmalignant tumor cells (heterotypic fusion). In this case, fusion is thought
to promote chemo-resistance or metastatic potential of these cells.
Macrophages seem to have distinct fusogenic properties which leadto physiological or pathological output. We hope to make advancesin understanding the mechanisms and molecules involved in cell-cellfusion. We are mainly focusing on homotypic cell-cell fusion such as
osteoclasts and multinucleated giant macrophages.

Noriko Takegahara, MD/PhD

To address fusion competent cells, we will focus on the close
relationship of cell development to cell cycle. Fucci is a powerful
technology developed by Dr. Atsushi Miyawaki to study coordinationof cell cycle and cell development. Using Fucci, we can investigate how
cell cycle regulates the setting of appropriate timing of generation ofdifferent cell types. We apply Fucci technology to find fusion competentcells in osteoclasts- and multinucleated giant macrophage-precursors(Figure 2). We are also trying to identify fusogens which regulate cell-cellfusion.
Noriko Takegahara

Assistant Professor

Kazuaki Tokunaga(Visiting Academic Staff)

Cell-cell fusion is a dynamic process. Various types ofmolecules change their expression levels and their cellular
localization during the process. Recent advances in opticalimaging technologies allow us to detect qualitative changes
over time. However it is still hard to detect quantitative changes
over time. In collaboration with Nikon Instruments Company,
we would like to develop a novel technology to quantify
changes of molecular levels in in vitro and in vivo images.

An approach to find fusion competent cells
Once myeloid precursors receive signals of differentiation factor (RANKLfor osteoclasts, IL-4 for multinucleated giant macrophages), they commit
to be pre-osteoclasts or pre-giant macrophages and fuse to become
mature multinucleated cells (Figure 1). Myeloid precursors need to passmultiple steps to become mature multinucleated cells and one of thesteps is to become fusion competent cells. Although it has been reported
that cell cycle arrest in osteoclast precursors is important for theirdifferentiation into multinucleated osteoclasts, the biology of cell-cellfusion, especially, how cells commit to become fusion competent status
and what molecules are actual fusogens remains to be determined.

Associate Professor

A challenge of development of a novel imaging technology

Figure 1. Schematic summary of myeloid cell fusion. Common myeloid
lineage precursors commit to becoming fusion competentstatus and then fuse to become mature multinucleated cells.

Figure 2. Fluorescence micrographs of Fucci-expressing monocytes
and osteoclasts. Cells expressing green fluorescence are in
S/G2/M phases while cells expressing red fluorescence arein G1 phase.

Recent Publications
 Kim H, Kim T, Jeong BC, Cho IT, Han D, Takegahara N, Negishi-Koga T,
Takayanagi H, Lee JH, Sul JY, Prasad V, Lee SH, Choi Y. Tmem64 modulates
calcium signaling during RANKL-mediated osteoclast differentiation. Cell
Metab. 17:249-60, 2013.

 Kang S, Okuno T, Takegahara N, Takamatsu H, Nojima S, Kimura T, Yoshida
Y, Ito D, Ohmae S, You DJ, Toyofuku T, Jang MH, Kumanogoh A. Intestinal
Epithelial Cell-Derived Semaphorin 7A Negatively Regulates Developmentof Colitis via αvβ1 Integrin. J. Immunol. 188:1108-16, 2012.

 Hirosue A, Ishihara K, Tokunaga K, Watanabe T, Saitoh N, Chandra T, Narita
M, Shinohara M and Nakao M. Quantitative assessment of higher-orderchromatin structure of the INK4/ARF locus in human senescent cells. Aging
Cell 11:553–56, 2012.

 Takegahara N, Kang S, Nojima S, Takamatsu H, Okuno T, Kikutani H,
Toyofuku T, Kumanogoh A. Integral roles of a guanine nucleotide
exchange factor, FARP2, in osteoclast podosome rearrangements. FASEB J.
24:4782-92, 2010.

 Kuwajima T, Yoshida Y, Takegahara N, Petros TJ, Kumanogoh A, Jessell TM,
Sakurai T, Mason C. Optic chiasm presentation of Semaphorin6D in thecontext of Plexin-A1 and Nr-CAM promotes retinal axon midline crossing.
Neuron 74:676-90, 2012.

58

59

Symposia & Seminars

60Symposia & Seminars

International Symposium TCUID
Speaker

Date:

November 18-20, 2013

Venue:

Icho Kaikan & Taniguchi Memorial Hall, Osaka University

The International Symposium “Towards Comprehensive
Understanding of Immune Dynamism” (TCUID 2013) was
held on the 18th-20th of November at the Icho-Kaikan, Osaka
University. This was the third and the final symposium for
AKIRA Project, and was supported by the Cabinet Officeand the Japan Society for the Promotion of Science with
176 participants in total. Eight speakers from IFReC andAKIRA Project, and nine invited speakers from four different
countries including the special lecturer presented theirlatest research findings. As for the poster session, there
were 28 presentations led by young researchers who saw
lively questions from the participants. In TCUID 2013, theparticipants could deepen their understanding of the
dynamic mechanisms regulating innate and acquired

62

immunity throughpresentations anddiscussions of researchfindings achievedby using a variet y oftechniques including
imaging and systems
biology.
A Young Researchers’
Workshop was also held on the first day, where young
researchers from various areas of specialization were divided
into smaller groups to have group discussions, which
promoted exchange among the participants for futureinterdisciplinary researches.

Title

Michael Reth (Keynote Lecturer)(University of Freiburg, Max Planck Institute of
Immunobiology and Epigenetics, Germany)

Mapping the Lymphocyte Surface: a Nanoscale Study

Myung-Shik Lee(Sungkyunkwan University School of Medicine, Korea)

The Zinc Finger Domain Containing Nuclease, Regnase-1/Zc3h12a, Controls T
Cell Activation

Thirumala-Devi Kanneganti(St. Jude Children’s Research Hospital, USA)Mediators of Inflammatory Responses

Kensuke Miyake(The University of Tokyo, Japan)

Mechanisms Regulating Nucleic Acid Sensing Toll-like Receptors

Masahiro Yamamoto(Osaka University, Japan)

Toxoplasma gondii Polymorphic Dense Granule Protein GRA6 Selectively andStrain-specifically Activates NFAT4

Cevayir Coban(Osaka University, Japan)

A New Gateway for Cerebral Malaria Immunopathology

Kenta Nakai(The University of Tokyo, Japan)

Analyses of Transcriptional Changes over Time in Stimulated Dendritic Cells
through Systems Biology

Grégoire Altan-Bonnet(Memorial Sloan-Kettering Cancer Center, USA)

From Local Signals to Global Responses: Self-organization of T- Lymphocyte
Activation through Cytokine Communications

Fuyuhiko Inagaki(Hokkaido University, Japan)

Structural Biology of Innate Immunity

Stefan J. Riedl(Sanford-Burnham Medical Research Institute, USA)

Mechanism and Regulation of Cell Signaling by Death Domains

Masaru Ishii(Osaka University, Japan)

Dynamic Behavior of Osteoclasts and Their Precursors Macrophages Visualized
by Intravital Microscopy

Joji Fujisaki(Columbia University, USA)

Immune Privilege of the Hematopoietic Stem Cell Niche

Nicholas I. Smith(Osaka University, Japan)

Label Free Measurement of Immune Cells: Inflammasome Induction and RNA
Molecular Analysis

Kazuya Kikuchi(Osaka University, Japan)

In Vivo Imaging by Designed MRI Probes with Tunable Chemical Switches

Erik M. Shapiro(Michigan State University, USA)

Monitoring Immune Cell Infiltration Using MRI-based Cell Tracking

Mark S. Sundrud(The Scripps Research Institute, USA)

Dynamics and Molecular Features of Inflammatory T Cell Subsets in Human
Autoimmune Disease

Shizuo Akira(Osaka University, Japan)

NLRP3 Inflammasome and Microtubule Acetylation

63

Symposia & Seminars

Immunology Frontier "B to B Seminar"

Mini Symposium on Malaria Immunopathology
Date:

January 15, 2014

Venue:

Meeting Room #1, IFReC Research Building

Speaker:

Laurent Renia (Singapore Network-BMSI-A* STAR)
Breadth of the humoral response and potential
vaccine targets following experimental Plasmodium
falciparum sporozoite inoculation in humans
Katsuyuki Yui (Nagasaki University)

M alaria

ini symposium on

Immunopathology
January 15, 2014

Meeting room #1, IFReC Bldg. 2F
Speakers:

13:30-14:10 Laurent Renia (Singapore Network-BMSI-A* STAR)
Breadth of the humoral response and potential vaccine targets following
experimental Plasmodium falciparum sporozoite inoculation in humans
14:10-14:50 Katsuyuki Yui (Nagasaki University)
T cell responses during infection with Plasmodium berghei ANKA

T cell responses during infection with Plasmodium

14:50-15:10 Coffee Breakberghei ANKA

15:10-15:50 Richard Culleton (Nagasaki University)
Evolutionary origins of P. falciparum and P. vivax as human pathogens

Richard Culleton (Nagasaki University)
Evolutionary origins of P. falciparum and P. vivax ashuman pathogens
Cevayir Coban (Osaka University)

15:50-16:30 Cevayir Coban (Osaka University)
Imaging cerebral malaria

Malaria

Host : Malaria Immunology
http://malimm.ifrec.osaka-u.ac.jp/

Immunopathology

Imaging cerebral malaria
Host:

The Immunology Frontier "B to B (Bench to Bed, Bed to
Bench) Seminar" was organized by Prof. Atsushi Kumanogoh(Osaka University School of Medicine, IFReC) and other IFReC
PIs. In the meeting, over 150 audience members had the plea-

Date:

May 24, 2013

Venue:

Icho Kaikan, Osaka University

Speaker:

Shimon Sakaguchi (Experimental Immunology, IFReC)
Kazuhiko Yamamoto (Department of Allergy and Rheumatology,
The University of Tokyo Hospital)

Laboratory of Malaria Immunology, IFReC

sure to listen to "Regulatory T cell and autoimmunity; the basis
for cancer immunology" by Shimon Sakaguchi and "The recent
breakthroughs in the treatment of rheumatoid arthritis andhuman immunology" by Kazuhiko Yamamoto.

Kishimoto Foundation Lecture
Date:

October 8, 2013

Venue:

Taniguchi Memorial Hall, Osaka University

Speaker:Prof. David Isenberg (Academic Director and Arthritis Research
UK Diamond Jubilee Professor of Rheumatology, University
College London)

64

Title:

Systemic Lupus Erythematosus -Outcome and Origins

Host:

Research Institute for Microbial Diseases & IFReC

65

Symposia & Seminars

IFReC Seminars
FY2013 IFReC Seminar
Date

Speaker

Affiliation

Title

2013
May 9

Noriko Takegahara

Department of Pathology and Laboratory Medicine,
University of Pennsylvania School of Medicine

Elucidation of osteoclast fusion mechanisms

May 16

Maho Hamasaki

Department of Genetics,
Graduate School of Medicine, Osaka University

Autophagosome membrane biogenesis

Jun. 4

Axel Kallies

Division of Molecular Immunology,
Walter and Eliza Hall Institute of Medical Research

IRF4 is a master regulator of T cell differentiation and metabolism

Jun. 17

Gos Micklem

Department of Genetics, University of Cambridge

From model organisms to human disease
and back again: data warehousing formodern biology

Jul. 18

Susana Guerra

Department of Preventive Medicine and Public Health,
Universidad Aut.ANsnoma, Madrid.

ISG15 regulates peritoneal macrophages
functionality against viral infection

Dec. 9Takeshi Egawa

Transcription factors regulating duration of T
Pathology and Immunology,
cell immune responses and host protection
Washington University in St. Louise, School of Medicine
through metabolic regulation

Dec. 24

Florent Ginhoux

Singapore Immunology Network

Dendritic Cells and Macrophages in Healthand Disease

Jan. 15

Laurent Renia

Singapore Immunology Network

Breadth of the humoral response and potential vaccine targets following experimental
Plasmodium falciparum sporozoite inoculation in humans

Jan. 15

Katsuyuki Yui

Graduate School of Biomedical Sciences, Nagasaki University

T cell responses during infection with Plasmodium berghei ANKA

Jan. 15

Richard Culleton

Institute of Tropical Medicine, Nagasaki University

Evolutionary origins of P. falciparum and P.
vivax as human pathogens

Jan. 15

Cevayir Coban

Malaria Immunology, IFReC

Imaging cerebral malaria

Jan. 30

Eicke Latz

Institute of Innate Immunity, BiomedicalCenter,
University of Bonn
Division of Infectious Diseases & Immunology,
University of Massachusetts Medical School

Mechanisms of Sterile Inflammation

Feb. 4

Shan Lu

Department of Medicine University of Massachusetts
Medical School

Heterologous DNA prime-protein boost HIV
vaccines

2014

IFReC holds seminars throughout the year with speakers from a variety of disciplines including immunology, imaging and informatics with the aim of promoting
collaborative research, as well as to inspire and educate the next generation ofscientists.

IFReC Seminar“Mechanisms of Sterile Inflammation”

Eicke LATZ, MD, PhD
Institute of Innate Immunity, Biomedical Center,
University of Bonn, Germany /Division of Infectious Diseases &Immunology,
University of Massachusetts Medical School, USA

DATE Thursday, January 30, 2014
TIME 11:00 am - 12:00 pm
VENUE Taniguchi Memorial Hall, Integrated Life Science Bld. 1F
HOST Ken Ishii (Laboratory for Vaccine Science, IFReC)

66

67

Symposia & Seminars

IFReC Colloquia
IFReC started a series of colloquia for IFReC researchers in
FY2011. Researchers from all labs are able to present and discuss their current research progress. These events were initiated as a means to make all those present at IFReC aware of thewealth of immunological research that is being conducted,
and to promote collaborations between all the immunology,
imaging and informatics groups. An average of approximately
100 people attend each time.

Date:

The 9th IFReC Colloquium: April 24, 2013
The 10th IFReC Colloquium: June 12, 2013th

The 11 IFReC Colloquium: August 7, 2013
The 12th IFReC Colloquium: October 16, 2013
The 13th IFReC Colloquium: December 18, 2013
The 14th IFReC Colloquium: February 12, 2014
Venue:

Speaker
Syoji Kobashi (Information Systems)

Soft computing for image processing to quantify in-vivo images

Kei Kuramoto (Information Systems)

Simulation and soft computing for asthmatic attack

Hui Jin (Immunochemistry)

Misfolded ER proteins transported to the cell surface by MHC class II molecules are targeted byautoantibodies

Hiroshi Ike (Cell Signaling)

Fluctuation of DAG in immunological synapse regulates T cell activation

Kouji Kobiyama (Vaccine Science)

Targeting lymph node macrophage and dendritic cells by nano-particulated TLR9-ligand

Hong Zhao (Malaria Immunology)

Immunopathology of cerebral malaria: A new gateway

Tomokazu Ohta (Immune Regulation)

Homeostatic roles of XCR1-expressing dendritic cells in the intestinal immunity

Yoshiko Murakami(Immunoglycobiology)

Genetic and biochemical basis of inherited GPI-anchor deficiencies

Shinichiro Watanabe(Nuclear Medicine)

Whole-body glucose metabolism in identical twin pairs:Effects of genetic and acquired factors on variousorgansYuko Kamikawa(Chemical Imaging Techniques)

Development of PYP-based protein tag system for fluorogenic imaging of cellular proteins

Takeharu Minamitani(Molecular Immunology)

Epstein-Barr Virus LMP2a reduces the stringency of germinal center B cell selection: an implication for
persistent virus infection

Yoshihiro Oka (Immuno-pathology)

Cancer antigen WT1-targeting immunotherapy for patients with malignancies

Nam Trung Nguyen(Immune Regulation)

The roles of aryl hydrocarbon receptor in immune responses

Taisuke Nakahama(Immune Regulation)

Aryl hydrocarbon receptor-mediated induction of the microRNA-132/212 cluster promotes TH17 celldifferentiation

Masanori Matsumoto(Lymphocyte Differentiation)

Plasmablasts control T-cell autoimmunity through IL-10 production

Tomoyuki Yamaguchi(Single Molecule Imaging)

Models of T cell-mediated immune response

Yoko Yano (BioLegend Japan KK)

BioLegend collaboration strategies in Japan

Takatoku Oida (BioLegend Japan KK)

BioLegend launched Japan R&D lab at Saito

Keizo Nishikawa(Immunology and Cell Biology)

Dnmt3a as a novel nexus coordinating metabolism and epigenetics in osteoclasts

Atsushi Tanaka(Experimental Immunology)

Breakdown of immune tolerance by quantitative and qualitative alterations of TCR signaling through ZAP-70

Shide Liang (Systems Immunology)

Development of knowledge-based energy functions and applications in immunology research

Miwa Sasai (Immunoparasitology)

The Toxoplasma virulence effector GRA6 is required for selective and strain-specific NFAT4 activation

Masaki Fukunaga(Biofunctional Imaging)

Manganese enhanced MRI revealed microglia activation in the demyelinating disease model brain

Takamasa Ishidome (Immune Network)

Phagocytic clearance of live T lymphocytes in inflammation

Taniguchi Memorial Hall, Osaka University

Chair : Hironori Kawahara

14th
IFReC
Colloquium

3:25 pm|
4:00 pm“The Toxoplasma virulence eﬀector GRA6
is required for selective and strain-speciﬁc
NFAT4 activation”
Miwa Sasai /Immunoparasitology

Chair : Masahiro Yamamoto
Date

Wed., Feb. 12, 2014

4:00 pm|
4:35 pm“Manganese enhanced MRI revealed
microglia activation in the demyelinating
disease model brain”
Masaki Fukunaga / Biofunctional Imaging

4:35 pm|
5:10 pm“Phagocytic clearance of live T lymphocytes
in inﬂammation”
Takamasa Ishidome / Immune Network

5:10 pm|

Happy Hour

Venue

Taniguchi Memorial Hall

Please note that the starting time of
the 14th colloquium is3:25pm.

“IFReC Colloquium” is the seminar series open to IFReC members only.
At each seminar, speakers from IFReC laboratories will talk about their recent
research progress.

Title

Please do not disclose what you hear in the seminar to outside parties, because
each presentation contains unpublished data.
Happy Hour will be held after the colloquium to enhance exchanges between
IFReC members.

68

69

Live Immuno-Imaging Facility Opening Workshop

EventsIFReC established the “Live Animal
Immuno-imaging Facility” in April, 2013.
To commemorate its opening, the "Live
Immuno-Imaging Facility" Opening
Workshop was held. At the beginning,
Yoshichika Yoshioka (PI, Biofunctional
Imaging Lab, IFReC) gave a lecture titled“Live Immuno-Imaging: From small animal to human.” In the workshop, sevenresearchers presented their current
research progress related to bioimaging
technology. Afterwards, a tour to the facility equipped with an 11.7T MRI and amulti-photon fluorescence microscope
was arranged for the participants.

Date:

June 19, 2013

Venue:

Taniguchi Memorial Hall, Osaka University<Keynote Lecture>
15:00 -15:30 Yoshichika Yoshioka (PI, Biofunctional Imaging Lab, IFReC)
Live Immuno-Imaging: from Small Animal to Human<Morphological Studies>
15:30 -15:40 Masaaki Murakami (Developmental Immunology)
15:40 -15:50 Yuki Mori (Biofunctional Imaging)<Functional Studies>
15:50 -16:00 Hiroyuki Murota (Dept. Dermatology, School of Medicine)
16:00 -16:10 Masaki Fukunaga (Biofunctional Imaging)<Cellular-tracking Studies>
16:10 -16:20 Manabu Kinoshita (Dept. Neurosurgery, School of Medicine)
16:20 -16:30 Takashi Jin (RIKEN QBiC)
16:30 -16:40 Yoshikazu Tsukasaki (RIKEN QBiC)
16:40 -17:00 Questions and Answers

70Events

Winter School
Lecturer

Title

Hisashi Arase
WPI Immunology Frontier Research Center (IFReC), Regulation of Immune Response by Paired Receptors
Osaka University, JapanThe third Winter School on Advanced Immunology was jointly organized with
Singapore Immunology Network (SIgN).
Forty eight young researchers, who were competitively selected from 217 applicants, and 15 world leading immunologists got together on Awaji Island inJapan on 19-23 January 2014.
They shared intriguing insights and findings in immunology, discussed new
ideas and forged friendships that fueled networking and future collaborations.
The participants enjoyed a one-day tour to IFReC in Suita Campus of Osaka
University after the school in Awaji Island.

72

Date:

January 19-23, 2014

Venue:

Awaji Yumebutai International Conference Center, Awaji Island, Hyogo

Robert Brink
Garvan Institute of Medical Research, Australia

Unravelling the Complexities of the Germinal Centre Reaction

Marco Colonna
Washington University School of Medicine, USA

Innate Lymphoid Cells in Mucosal Immunity

Florent Ginhoux
Singapore Immunology Network

Dendritic Cell and Macrophage Immunobiology

Steffen Jung
Weizmann Institute of Science, Israel

Probing Macrophage Functions in Gut and Brain

Hajime Karasuyama
Tokyo Medical and Dental University, Japan

Emerging roles for basophils in immunity: a neglected minority gains new respect

Maria A. Curotto de Lafaille
Singapore Immunology Network

IgE and the Pathogenesis of Allergic Diseases

Andreas Radbruch
German Rheumatism Research Centre (DRFZ)

The Resting and the Restless Immunological Memory

Federica Sallusto
Institute for Research in Biomedicine, Switzerland

Effector and memory T cells in humans

Yousuke Takahama
University of Tokushima, JapanT cell repertoire formation in the thymus

Masahiro Yamamoto
A mechanism of IFN-γ-induced cell-autonomous immunity against an intracellular
WPI Immunology Frontier Research Center (IFReC),
protozoan parasite Toxoplasma gondii
Osaka University, Japan
Lucia Mori
Singapore Immunology Network (SIgN), Singapore

Lipid Antigen Presentation to T Cells

Sho Yamasaki
Kyushu University, Japan

Recognition of pathogens through C-type lectin receptors

Carola Vinuesa
The Australian National University

Multiple mechanisms operate to regulate T cell help for B cells

Christian Münz
University of Zurich, Switzerland

Macroautophagy in MHC restricted antigen presentation73

Events

Staff and Researchers Development Seminars

RDSD

Date:

August 9, 2013

Date:

February 5, 2014

Lecturer:

Mitiko Go

Lecturer:

Junko Tanaka(External Executive Director, Research Organization(Professor, Graduate School of Biomedical Sciences,of Information and Systems)

Hiroshima University)

Researcher and Staff Development

FY2013 “Being a Scientist” RD/SD Seminar Series*These seminars will be conducted only in Japanese

IFReC offered various Staff and Researcher Development programs in FY2013 to create
a motivated, skilled, and well-trained staff.
WPI拠点としての国際化・異分野融合を志向する教職員の意識改革を進めるために、
IFReC はRD/SDを積極的に展開します。
テーマは 「キャリアのターニングポイントにおいてどのように考え決断してきたか？」

Berlitz-IFReC Communication Course●第１回

The Berlitz-IFReC Communication Course is a new SD/RD program for IFReC members to
improve their English skills, communication and practice in the workplace. The 31 participants were selected by interview to join the course.

中村 桂子（NAKAMURA Keiko）

ＪＴ生命誌研究館館長【専門：生命基礎論（複雑系）・カオス・非平衡現象論】
日時：6月26日(水）：13時10分出発（IFReC）～16時10分到着（IFReC）予定
場所：JT生命誌研究館(高槻市） * 開催場所にご注意ください 。
交通：貸切バス
中村館長による講演30分/質疑応答15分/見学45分●第２回

原田 慶恵（HARADA Yoshie）

京都大学 iCeMS 教授 【専門：一分子計測・生物物理学】
日時：7月3日（水）：13時30分～15時30分
場所：谷口記念講堂
講演30分/質疑応答15分
若手研究者とのディスカッション （定員：10-15名）＊1時間程度 （IFReC棟：5階ラウンジ-事前申込み要/定員制)●第３回

Other Seminars for Staff Development
The following two seminars were held for administrative
staff to further promote expert knowledge. IFReC plans tocontinue the “Immunology introductory course series foradministrative & technical staff” during and beyond 2014
in collaboration with various scientists.

郷 通子（GO Michiko）

大学共同利用機関法人/情報・システム研究機構理事【専門：生物学・生物物理学】
日時：8月9日（金）：13時30分～15時30分
場所：谷口記念講堂
講演30分/質疑応答15分
若手研究者とのディスカッション （定員：10-15名）＊1時間程度 （IFReC棟：5階ラウンジ-事前申込み要/定員制）
企画室（内線４７７７）

Distinguished Women Scientist Seminars
One of the goals for IFReC and the university is to attract and recruit the best young researchers. Four distinguished women scientists were invited to IFReC to present about their academic career path and development. Through the series, about 200 participantsfrom both within and outside the university had the privilege to listen the lectures followed by a Question and Answer session.
Practice Theory for Outreachin the University-Know-how at IFReC-

Jun SAKANOUE(Press Information Officer of IFReC)

DATE
December 3, 2013
3:00 - 4:00 pm

VENUE
Meeting Room #1
IFReC Building

CONTACT: Jun SAKANOUE
j-sakano@ifrec.osaka-u.ac.jp

Date:

June 26, 2013

Date:

July 3, 2013

Lecturer:

Keiko Nakamura

Lecturer:

Yoshie Harada(Director, JT Biohistory Research Hall)
74(Professor, WPI-iCeMS, Kyoto University)

Date:

December 3, 2013

Date:

February 20, 2014

Title:“Practice Theory for Outreach in the University;

Title:“Immunology Introductory Course Series for

Know-How at IFReC”
Lecturer:

Jun SakanoueAdministrative & Technical Staff” Part 1
Lecturer:

Jun Sakanoue(Research Planning & Management Office,(Research Planning & Management Office,

IFReC)

IFReC)
75

Events

Japanese Lessons

August 8, 2013/ February 13, 2014

November 14, 2013

ORIGAMI

Let’s make plans for your virtual travel with a Japanese map !

What’ s the difference ?

ころころ。 ごろごろ。
とろとろ。 どろどろ。

Unlimited for the

IFReC Japanese Café

will be started from April 4.

IFReC
Japanese
Café
IFReC
Japanese
Café
IFReC Japanese Café
IFReC Japanese Café
IFReC Japanese Café
From April
4 to September
19 CaféIFReCJapanese

Do Japan!

Date & Time

Every Thursday, 18:30-20:00

IFReC Japanese Class
Registration required※Every Tuesday Evening, 18:30-20:00
From April 2, 2013 to September 17, 2013
(Up to 15 people, 20 Lessons, break in August)Eligible person

Intermediate and advanced Japanese Speakers※ Grammar, Hiragana, Katakana, etc. aiming at havingbasic knowledge necessary for conversation in daily life

OutlineThis year, interesting
themes related to Japan
were set in the Japanese
C a f e , a n d in t e r a c t i v elessons were conducted
through the themes ofJapanese traditional culture and trends. Japanese volunteers joined and helped thelessons to give foreign people an opportunity to practice in Japanese.

■Class B : For Elementary to Pre-intermediate

This is an actual opportunity to speak Japanesein casual style intended for intermediate, advanced
Japanese speakers. A certain topic/activity will begiven by a Japanese teacher in each session. You canbring your questions regarding Japanese language todiscuss with a Japanese teacher or other people.

※Every Wednesday Evening, 18:30-20:00

From April 3, 2013 to September 18, 2013
(Up to 15 people, 20 Lessons, break in August)※ Grammar, Phrase, case study etc. aiming at having
more practical knowledge for conversation in daily life

Free of charge for lesson

Do Japan!

76

きりきり。 ぎりぎり。

■Class A : For Absolute Beginner

2F Refresh Room, IFReC Bldg.

We offer "Class A: Absolute beginner class", "Class B: Elementary/Pre-intermediate Class" held in lecture style to learn basic
knowledge necessary for conversations, and "Japanese Cafe
: Intermediate/advanced class" held in a cafe style to learn
more practical phrases and idioms in daily life from a professional Japanese teacher.

ふつふつ。 ぶつぶつ。

--- First Semester ---Venue

We hold Japanese language classes for overseas researchers / students to help reduce the stress and inconvenience that are caused by
the language barrier in a research environment as well as in daily life.

へとへと。 べとべと。

e
tim
st

First Semester

きらきら。 ぎらぎら。

ﬁr

Required to purchase text books

July 18, 2013

September 12, 2013

January 16, 2014

Osakan culture “TAKOYAKI”

SHODO

The first writing of the New Year (KAKIZOME)

October 31, 2013
Let’s make an ONIGIRI ! (Rice Ball)

December 12, 2013

March 13, 2014

Let’s try to write a New Year’s greeting card by using
a fude (a brush ) !

Let’s introduce your country with a world map!

77

Outreach Activities

78

Outreach Activities

Science Café Series “Café on the Edge”

Super Science High School Festa 2013

IFReC and FIRST Program AKIRA Project have organized “Café on the Edge”
since FY2010. We held “Café on the Edge” two times in FY2013 with the speakers
chosen from cutting-edge science fields of IFReC. The number of participants
totaled over 150.

Date:

May 3, 2013

Venue:

Techno Alliance Building, Osaka University

Guest:

Yoshichika Yoshioka (Biofunctional Imaging, IFReC)

Title:“Toward the fullest potential of MRI; Neuron activities toimmune reactions”

The Purpose of the project for Super Science High School (SSH) is to promote
upper secondary education to foster excellent human resources in scienceand technology. The SSHs are designated to be developed enriched curricula,teaching methods, and materials for science and mathematics in cooperation
with universities and research institutes.
OL
HO
IFReC and other WPI institutes participated in the annual meeting of SSH 2013
SC
H
G
in Yokohama. We presented the research fields conducted at IFReC, and hope CE HINECI
the information could help students to choose a university in the near future.
RS

R E
OL
PE
SU IENC CHO
SC GHS
HI

定

省指

科学

文部

14

サ

ー

パ

ー

ス

エ

イ

ル

ー

ク

ス

イ

ハ

ス

ン

20

〜
13

20

E

Date:

June 12, 2013

Venue:Art Area B1 (Naniwabashi station, Osaka)

Guest:

Osamu Takeuchi

Title:

80

P
SU(Institute for Virus Research, Kyoto University & IFReC)

Date:

August 7 & 8, 2013

“Inflammation; Us-or-them?”

Venue:

Pacifico Yokohama, Kanagawa

Host:JST, MEXT

81

Outreach Activities

Science Agora 2013

Science Talk Live 2013 by WPI“Science Agora” is an event designed based on “Family Science Day” atannual meeting of American Association for the Advancement of Science. In
the “Science Agora 2013” in Tokyo, WPI institutes co-organized “Science Talk
Live” for kids and general citizens. After the event, we were awarded “Science
Agora Prize” for the introduction of the most-advanced science and technology in Japan.

Date:

November 9 & 10, 2013

Venue:

National Museum of Emerging Science and Innovation, Tokyo

Host:

JST

This event started in 2011 to encourage a close relationshipbetween WPI scientists and high school students. The third
symposium “The perceptive scientists change the World!” washeld in Sendai. In the event, the researchers from WPI insti-

tutes talked about their cutting-edge researches, and spokeon passion for science. At the end of the event, students andscientists spoke freely with one another and enjoyed experiments at each WPI booth.

Date:

December 14, 2013

Venue:

Sendai International Center, Miyagi

Host:

Advanced Institute for Materials Research, Tohoku University (WPI-AIMR), WPI Institutes

Support:Miyagi Prefecture, MEXT, JSPS

Speaker Kenichiro Itami (Director of WPI-ITbM, Nagoya University) Kei Hirose (Director of WPI-ELSI, Tokyo University of Technology)
 Masashi Yanagisawa (Director of WPI-IIIS, University of Tsukuba) Alan Greer (Principal Investigator of WPI-AIMR, Tohoku University, Professorof University of Cambridge)
 Akari Takayama (JSPS Postdoctoral Fellow, WPI-AIMR, Tohoku University)

Presentations by Students
Sendai Dai-ichi High School / Sendai Dai-san High School / Furukawa Reimei
High School / Eleanor Roosevelt High School, USA

82

83

Outreach Activities

Symposium for General Audience "Future Medical Treatment Created by Immunology"

AAAS 2014
The American Association for the Advancement of Science(AAAS) is the biggest international non-profit organization for
the advancement of science in the world. The main theme forthe recent annual meeting of AAAS (AAAS 2014) was "Meeting
Global Challenges: Discovery and Innovation".
In the exhibition, as part of the Japan pavilion at AAAS 2014,
the Press Information Officers from all the WPI institutes introduced the collaborative researches bridging various research

Date:

February 1, 2014

Venue:MIRAICAN Hall, National Museum of Emerging Science and Innovation, Tokyo

Host:

IFReC, FIRST Program AKIRA Project, and the Mainichi Newspapers Co.

Date:

February 13-17, 2014

Venue:

Hyatt Regency Chicago, IL, USA

fields in an internationally opened environment of each
institute. The Japan pavilion received the greatest numberof guests among all the booths. The WPIs and RIKEN co-organized the exhibitor workshop "Build a Career in Japan!"on Feb. 14. In the workshop, we provided an opportunity fordiscussing Japan's science policy and career path for overseas
researchers.

<Program>
Lecture I "Potential of New Treatments Created by Immunology"
Shizuo Akira (Director of IFReC)
Lecture II "Antibody Drugs- The Potential and Prospect"
Kunihiro Hattori (Chugai Pharmaceutical Co., Ltd.)

Talk Session "Future Medical Treatment Created by Immunology"
 Shizuo Akira (IFReC)
 Kunihiro Hattori (Chugai Pharmaceutical Co., Ltd.) Tomohiro Kurosaki (IFReC)
 Junichi Taguchi (Hospital Director, Tokyo Midtown Clinic)
 Yuri Kato (Personality)
 Yukiko Motomura (the Mainichi Newspaper)

84

内閣府

独立行政法人日本学術振興会

85Research Projects

86Research Projects

Research Support Program for Fusion of Different Fields
The final goal of the World Premier International ResearchCenter (WPI) Initiative program is to create world leading
research centers where top researchers gather, enabling theachievement of high-level research results, and at the sametime, providing an excellent research environment. To achieve
one of the goals of the WPI program, each center is expected
to generate novel research fields through the promotion of
combined research projects.

Here at IFReC, we are aiming to create innovative immunology fields by combining with imaging and bioinformatic technologies. In order to support this new challenge, we established the Research Support Program for Combined Research
Fields in FY2009 for IFReC members. A total of 10 combined
research projects and one Dual Mentor Program are currently
underway.

Projects Supported by the Research Support Program for Fusion of Different Fields
Date
Started
Oct. 2012

Oct. 2013

Principal
Researcher

Collaborator

Titles of Proposed Project

M Yamamoto

D StandleyEva-Maria Frickel

Trilateral analysis of interferon-γ-mediated cellular innate immunity against
Toxoplasma gondii

F Sugihara

R Hanayama
K Kikuchi

in vivo Imaging of Germinal Center Development in Mouse Spleen Using MRI

B Ripley

D Standley
G Kurisu

Role of Arid5A in the Selective Control of IL-6 mRNA Stability and Development of
TH17 Cells

M Kohyama

C CobanK SuzukiRole of tissue macrophage in malaria infection, and their developmental control byparasite metabolite

T Yamaguchi

H Fujita
S Sakaguchi
T Watanabe
T Jin

Imaging analysis of Imuune activation and regulation

Y Kumagai

J Kozuka
S Teraguchi
N TrostVisualizing Information Processing of Immune Cells via Combination of Fluorescent
Reporter and Single Molecule Imaging

K Suzuki

Y BabaVisualizing activation of germinal center B cells using genetically encoded calcium indicators

N Ohkura

A Vandenbon
S Nakamura
S Yamazaki
S KatoM Hashimoto

Development of an epigenome-based computational classification system for thetreatment of autoimmune diseases

A Vandenbon

S Sakaguchi
H Morikawa
N Ohkura

Identification of key factors for inducing functionally stable regulatory T cells

D DiezR Hanayama

The dynamics of novel signaling networks of macrophages exposed to pathogens

Researchers from the groups of <Immunology>, <Imaging>, <Informatics>, and <Other>
88

89

Research Projects

Combined Research Fields Evaluation Workshop
The evaluation workshop was held on September 19, 2013
at Taniguchi Memorial Hall. Both the Combined Research Program and Dual Mentor Program recipients for FY2012 made a
research-in-progress presentation for 20 minutes. All IFReC PIs
joined the workshop as reviewers. The evaluation workshop

Date:

September 19, 2013

Venue:

Taniguchi Memorial Hall

provides opportunities for the promotion of interdisciplinary
research activities among young researchers, introducing thecurrent research at IFReC. The newly selected recipients of the
program for FY2013 gave two-minute presentations to introduce their research.

Dual Mentor Program
The Dual Mentor Program aims to help young researchers
engage in inter-disciplinary research by providing them with
two advisors and/or mentors with specialization in different
research fields. The annual budget for each project is ¥3 mil-lion with an internal evaluation once a year. The program wasintroduced as a platform to further promote interdisciplinary
research at IFReC.

Takeshi Yoshida (C) and his two mentors, Rikinari Hanayama (L) and Kazuhiro Suzuki (R).

Evaluation Criteria Chart

Recipient
T YOSHIDA(Immune Network)

MentorR Hanayama (Immune Network)
K Suzuki(Immune Response Dynamics)

Project Title
Visualizing the dynamics of exosomes during various
immune responses in vivo

Young Scientist Support Program for Research Abroad
To strengthen our international research network and ourbasis for international collaborative research, IFReC has provided financial support to young researchers who wish to participate in research activities at overseas institutions. The pro-gram aims to develop the practical skills and abilities of young
researchers towards international collaborative research andto develop their networking skills with researchers overseas.
Eleven researchers used this support program in FY2013.

Both document and presentation evaluations are made based on each of the elements aboveby all principal investigators. The results may determine whether the research grant amountwill be increased or decreased in the following year.

9091Research Projects

FIRST Program : AKIRA Project

The Funding Program for World-Leading Innovative R&D on
Science and Technology, (FIRST Program) AKIRA Project "Comprehensive understanding of immune dynamism: toward
manipulation of immune responses" led by core researcher,
Shizuo AKIRA and supported by JSPS and the Cabinet Office, Government of Japan, finished successfully at the endof March, 2014. The AKIRA Project produced a lot of fruitfulresults from the intensive research during the past four years,
including interdisciplinary research between immunology,
imaging, structural biology and systems biology, and the results were published as approximately 100 papers in journals
with high impact factors. During the four years, 13 invention
reports were submitted, and one was patented.

AKIRA Project made a DVD movie as a final summary of the
project and introduced the following four outstanding research results in it. DVDs were distributed at several outreach

events and a short version of the movie is planned to be madeavailable on our website.

Regulation of inflammation
by Regnase-1

Formation and activation of the
NLRP3 inflammasome

Regnase-1 regulates the inflammation by degradationof mRNA. Dynamic control of Regnase-1 expression
is critical for controlling activation of innate as well as
acquired immune cells.

The molecular mechanism of gout in immune cells was
elucidated. Microtubule-driven spatial arrangementof mitochondria promotes activation of the NLRP3
inflammasome.

Activities of the AKIRA Project in FY2013 (Except for outreach activities)
Date

Activity

2013
Apr. 16

The Fourth Patent Committee Meeting of the AKIRA Project

Apr. 24

The Fourth Meeting of 30 Operational Support Institutions in the FIRST Program by Cabinet Office, Government of
Japan

May 1

Issue of Leaflet “Immunology On the Edge” (Revised third edition)(in Japanese)

Jul. 16

The Fifth Patent Committee Meeting of the AKIRA Project

Jul. 31

Issue of Leaflet “Immunology On the Edge” (Revised fourth edition) (in Japanese)

Aug. 20

Establishing the Website of Intellectual Property (IP) for AKIRA Project

Sep. 6The Sixth Patent Committee Meeting of the AKIRA Project

Sep. 27

The Fifth Meeting of 30 Operational Support Institutions in the FIRST Program by Cabinet Office, Government of
Japan

Oct. 1

Researchers’ Development Seminar “Science Communication Seminar” (in Japanese)

Nov. 18-20

International Symposium TCUID 2013 (including Young Researchers’ Workshop)

Nov. 20

The Sixth Steering Committee Meeting of the AKIRA Project, FIRST Program

Nov. 29

Researchers’ Development Seminar “Career Path Seminar” (in Japanese)

Jan. 17

Production of the DVD “Novel Development of Immunology Research” for introducing outcomes of the AKIRA Project(in Japanese)

Mar. 5-6

The AKIRA Project Research Meeting 2014

Mar. 6The Seventh Steering Committee Meeting of the AKIRA Project, FIRST Program

Mar. 28

Issue of Research Achievement Summary Report of the AKIRA Project, FIRST Program(FY2010-FY2013) (in Japanese)

Disease specific M2 Macrophages
S. AKIRA’s group made the world-first discovery thatmacrophages are categorized to disease specific classes,
and identified macrophages specific to metabolic
syndrome and allergies.

Visualization of
the dynamic conversion of
osteoclast function
K.KIKUCHI’s group and M.ISHII’s group have succeededin visualizing the dynamic conversion of the osteoclast
func tion using 2-photon microscopy and newly
developed chemical probes.

2014

92

93

Research Projects

Activities
In FY2013, the AKIRA Project actively engaged in more events
than ever before. We hosted five outreach activities, and opened
booths and/or gave lectures at five other events. In every event,
we tried to have better communication with the general public
in order to enhance people’s understanding of our project andits importance. In the last year of our project, we also held several seminars aimed at researchers’ development for their future
activities.

Activities for General Public
Science cafes, lectures and the symposium

Event

Date and Venue

Speaker

Panel exhibition and Science Café
On the Edge 12

May2-3
Osaka Univ.

Yoshichika Yoshioka

Science Café On the Edge 13,“Inflammation: Friend or Foe of Our Body?”

June 12Art Area B1

Osamu Takeuchi

FIRST/WPI Researcher live lectures!
Young×Young in the "FY2013 Congress ofSuper Science High Schools"

August 8
Pacifico Yokohama

Yutaka Suzuki

Lecture for public, “The Milky Way in our body~the function and the movement of immune
cells through two-photon live imaging~”

November 24‘Sophia Sakai,’Kyoikubunka Center(Center for Education & Culture),
Sakai City

Masaru Ishii and Joji Fujisaki

Lecture: OSAKA GAKUGEISECONDARY SCHOOL visit

December 16
Osaka Univ.

Masaru Ishii and Shizuo Akira

Symposium for General Audience, “Future Medical
Treatment Created by Immunology”

February 1
MIRAICAN Hall, National Museum of
Emerging Science and Innovation

Shizuo Akira,Tomohiro Kurosaki, andKunihiro Hattori(Chugai Pharmaceutical Co., Ltd.)

Lecture for Public at Planetarium

Researchers’ Development Seminars and Career Path Seminar
AKIRA Project held science communication seminars on Oct 1 (in Japanese)and on November 18 (in English, as young researchers’ workshop in TCUID
2013). Guest lecturers from the National Museum of Emerging Science andInnovation (Miraikan) talked about helpful skills to interpret one’s researchefficiently and effectively. A career path seminar was also held on November
29, in which Masaru Ishii and Joji Fujisaki (Columbia University, USA) talked
about their career progression and what were the key points for them.

94

Twinkling cells on planetarium dome - AKIRA Project held a public lecture, in which live cell imaging movies taken by
two-photon microscopy were projected on the dome screen of planetarium at ‘Sophia Sakai,’ on 24 November. Masaru Ishii,
one of the Sub-Project Leaders and Joji Fujisaki, an assistant professor of Columbia University, got on stage. Ishii led the eventand explained the contents of themovies. They also explained to theparticipants interesting aspects ofconducting researches, as well as what
inspired them to become scientists.

Other Open Public Events
We opened a booth at four open public events, “Science
Agora 2013 (in Tokyo)”, “FIRST Business Matching Symposium(in Sendai and in Kyoto)” and “FIRST EXPO 2014 (in Tokyo)”.
In addition, Core Researcher Shizuo Akira gave lectures andjoined panel sessions at “FIRST Business Matching Symposiumin Kyoto” and “FIRST EXPO 2014”. All the events gathered thousands of people in total ranging from kids to the elderly.

95

Data

96

Data

Facilities
cated in specific laboratories of IFReC or RIMD. Collaboration
is key to the goals of IFReC; all facilities are accessible to any
researcher, facilitated with help from the core facilities management group.

IFReC and the Research Institute for Microbial Diseases (RIMD)
Research Complex adjoins the Institute for Protein Researchand the Graduate School of Engineering. This research complex offers a wealth of facilities to the researchers of IFReC,
shared and operated in part with RIMD. Instruments are lo-

IFReC–RIMD Research Complex at Suita Campus of Osaka University

1
2
8

3

Animal Resource Center (A, B &C)
 Pathogen-free animal facilities
 Embryo freezing and preservation facilities
 Transgenic and knock-out animal generation facilities
 In vivo experiment facilities Basic biological experiment facilities

7
6

Immunology Groups DNA sequencers
 Cell sorters and flow cytometers Real-time PCRs
 Spectrometers
 Two-photon microscopes
 Super-high resolution microscope

5

4

Imaging Groups

Core Facilities with Skilled Technicians

 MRI (11.7T)
 TEM / Cryo TEM
 Raman microscopes

 Animal facility A (for infected animal experiments)
 Animal facilities B&C (specific pathogen-free) Radio isotope experiment facilities DNA chip facilities
 Next generation DNA sequencer DNA sequencers
 Cell sorters and flow cytometers
 TEM
 Mass spectrometers

 Two-photon microscopes
 Super-high resolution microscopes
 Total reflective microscopes
 Confocal microscopes
 Cryo-microtomes
 DNA sequencers
 NMR
 Mass spectrometers Spectrometers
 Chromatography (LC/HPLC/FPLC) Dynamic light scattering analyzer
 Biacores Peptide synthesizer

Photo: S. Higashiyama

1. IFReC Research Building

5. Cutting-edge Research Building for Infectious Diseases

2. Integrated Life Science Building

6. Animal Resource Centers for Infectious Diseases (Building A, B, C)

3. Research Institute for Microbial Diseases (RIMD)

7. Institute for Protein Research

4. The Genome Information Research Center, RIMD

8. Graduate School for Engineering

Informatics Groups Bioinformatics server

11.7T MRI
Preparation room

P2A

P2A

Animal RoomOperating Room*safety cabinet

MRI
MachineRoom

Ante-Room

Fluorescence
Microscope

Two Photon
Microscope

PB
ChangeRoomP.S.

SPF animal Area
Biohazard Area

The Live Animal Immuno-imaging Facility equipped withan 11.7T MRI and a multi-photon fluorescence microscope, isdesigned with a clean room adjacent to the laboratory. It is
98

These photos were taken before sterilization.

Technicians of Core Facilities
Facilities available to researchers at IFReC are owned, operated
and maintained by the management group for the core facilities, or individual laboratories, to allow researchers to focus
more on their work, various kinds of equipment are run andmaintained by skilled individuals providing a professional service.

All the staff of Core Facilities outside the RIMD and IFReC buildings

expected that this facility will make great contributions to thedevelopment of fusion research between immunology andimaging at IFReC.
99

Data

Kishimoto Foundation Fellowships

Members

The Kishimoto Foundation was established in 2008 in honor of Tadamitsu Kishimoto, who, during the 1980s and 90s,
elucidated the function of interleukin-6 (IL-6), a key moleculefor stimulating immune responses. He later developed theanti-IL6 receptor-based therapy, tocilizumab, to treat immune
disorders such as Castleman's disease or rheumatoid arthritis.

IFReC launched the Kishimoto Foundation Fellowship program for researchers in various fields of immunology in 2010.
The program is supported by the Kishimoto Foundation anddesigned to support overseas researchers in order to promoteand develop immunological research and international exchanges at IFReC. The fellowships are open to postdoctoral
researchers of non-Japanese nationality who seek to collaborate with IFReC researchers. The recipients are provided witha salary and an airfare to Japan.

Number of IFReC Staff
300

250
300
200
250
150
200
100

FY2013 Kishimoto Fellowship Recipients

100

Position of Recipient

Name (initials)

Nationality

Host researcher

Period150
50Specially Appointed Researcher

H. Z.

China

Coban

Jul. 1, 2010 Dec. 31, 2013

100

Specially Appointed Researcher

R. S.

India

MurakamiAug. 16, 2010 Aug. 15, 2013

50

Specially Appointed Researcher

C. T.

TaiwanKikutani

Apr. 1, 2011 Mar. 31, 2014

Specially Appointed Researcher

J. B.

India

Akira

Apr. 1, 2011 Mar. 31, 2014

Specially Appointed Researcher

S. L.

Korea

KishimotoApr. 16, 2011 Mar. 31, 2014

Specially Appointed Researcher

W. L.

China

KishimotoSep. 1, 2011 Jun. 30, 2013

Specially Appointed Researcher

I. B.

Tunisia

Hanayama

Jan. 16, 2012 Jan. 15, 2015

Specially Appointed Researcher

K. N.

India

KishimotoJun. 1, 2012 Sep. 15, 2013

Specially Appointed Researcher

P. D.

India

KishimotoNov. 1, 2012 Oct. 31, 2014

Specially Appointed Researcher

J. H.

China

Sakaguchi

Dec. 1, 2013 Nov. 30, 2014

Specially Appointed Researcher

D. M.

BritainKishimotoDec. 1, 2013 Mar. 31, 2015

Research Fellow

N. N.

Vietnam

KishimotoSep. 21, 2013 Dec. 17, 2013

Research Fellow

F. G.

FranceKaishoDec. 16, 2013 Jan. 18, 2014

Research Fellow

P. S.

Singapore

KaishoDec. 16, 2013 Jan. 31, 2014

0

0

200

Number of Researchers
150
200

100
150

50
100

500

0

101

Data

AwardsToshio Yanagida

Person of Cultural Merit

The Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan announced
that Toshio Yanagida, Deputy Director of IFReC
was named a Person of Cultural Merit on October25, 2013. MEXT commented that Yanagida wasawarded by his outstanding achievements in thestudies of fundamental structure of biological system through the developments of single molecular measurements techniques.
This is the second time for a scientist of IFReC to
be honored as a Person of Cultural Merit, since Shizuo Akira (Director of IFReC) was awarded in 2009.

The Scientific Award, The Japanese Society forBone and Mineral Research
Masaru Ishii

Young Investigator’s Award, The Japanese Medical
Association
Masaru Ishii

Toshio Yanagida The Honorary Fellow of the Physical Society of Japan (JPS)The 37 scientists who greatly contributed to the development of physics have been chosen for JPS honorary fellow. The first honoree in 1950 was Hantaro Nagaoka, a former President of Osaka University. The others include, Hideki Yukawa, Shinichiro Tomonaga,
Makoto Kobayashi, Toshihide Masukawa, and Yoichiro Nambu (Nobel Prize laureates).
Yanagida was chosen by his outstanding achievements in the studies of fundamental structure of biological system through thedevelopments of single molecular measurements techniques.

Best Contribution Award,
The 14th International Symposium on AdvancedIntelligent Systems

Bayer International Publication Award
Yuki Mori,
Masaaki Murakami,
Yasunobu Arima,
Dasong Zhu, and Yoshichika Yoshioka

Award for Young Scientists,
The Pharmaceutical Society of Japan

Tatsuya Saitoh

Yutaka Hata

The Commendation for Science and Technology by
the Minister of Education, Culture, Sports, Scienceand Technology, Japan

Inoue Research Award for Young Scientists

Koichi Suzuki Memorial Award,
The Japanese Biochemical Society

Kenta Maruyama

Tetsuya Hirata

Hisashi Arase

102

Osaka Science Prize

Young Investigator Award,
The Japanese Association of Cancer Immunology

Kazuya Kikuchi

Daisuke Sugiyama
103

Finance
Break down of total expenditure at IFReC

Research Outputs

104

Research Outputs

Selected Articles
S1P-mediated osteoclast precursor monocyte migration is a critical point of control in anti-boneresorptive actions of active vitamin D

Arid5a controls IL-6 mRNA stability, which contributes to elevation of IL-6 level in vivo
Proc Natl Acad Sci USA 110(23):9409-14, 2013.

Proc Natl Acad Sci USA 110(17):7009-13, 2013.
Junichi Kikuta, Shunsuke Kawamura, Fumie Okiji, Mai Shirazaki, Sadaoki Sakai, Hitoshi Saito, and Masaru Ishii

Kazuya Masuda, Barry Ripley, Riko Nishimura, Takashi Mino, Osamu Takeuchi, Go Shioi, Hiroshi Kiyonari, and Tadamitsu Kishimoto

Masaru Ishii’s group show that calcitriol, which is the hormonally active form of vitamin D, and its therapeutically usedanalog, eldecalcitol, inhibit bone resorption by modulating
this mechanism. Vitamin D analogs have been used clinicallyfor treating osteoporosis, although the mode of its pharmacologic action remains to be fully elucidated. In this study, theyfound that active vitamin D reduced the expression of S1PR2,
a chemorepulsive receptor for blood S1P, on circulating osteoclast precursor monocytes both in vitro and in vivo. Calcitriolor eldecalcitol-treated monocytoid RAW264.7 cells, whichdisplay osteoclast precursor-like properties, migrated readily
to S1P. Concordantly, the mobility of circulating CX3CR1+
osteoclast precursor monocytes was significantly increasedon systemic administration of active vitamin D. These results
show a mechanism for active vitamin D in controlling the
migratory behavior of circulating osteoclast precursors, andthis action should be conducive to limiting osteoclastic bone
resorption in vivo.

Posttranscriptional regulation of IL-6 has been largely
uncharacterized, with the exception of the ribonuclease
Regnase-1, which prevents autoimmunity by destabilizing
IL-6 mRNA. Kishimoto’s group identified AT-rich interactive
domain-containing protein 5A (Arid5a) as a unique RNAbinding protein, which stabilizes IL-6 but not TNF-α mRNA
through binding to the 3’ untranslated region of IL-6 mRNA.
Arid5a was enhanced in macrophages in response to LPS, IL1β, and IL-6. Arid5a deficiency inhibited elevation of IL-6 serum level in LPS-treated mice and suppressed IL-6 levels andthe development of TH17 cells in experimental autoimmune
encephalomyelitis. Importantly, Arid5a inhibited the destabilizing effect of Regnase-1 on IL-6 mRNA. These results indicate
Arid5a plays an important role in promotion of inflammatory
processes and autoimmune diseases.

In vivo S1PR2-mediated control of migration of osteoclast
precursor monocytes visualized using intravital multiphoton
imaging.

Poly IC Triggers a Cathepsin D- and PS-1-Dependent Pathway to Enhance Cytokine Production
and Mediate Dendritic Cell Necroptosis

Arid5a stabilizes IL-6 through binding to the 3’ untranslated
region of IL-6 mRNA.

Malt-1-induced cleavage of Regnase-1 in CD4+ Helper T cells regulates immune activation
Cell 153(5):1036-49, 2013.

Immunity 38(4):717-28, 2013.
Jian Zou, Taro Kawai, Tetsuo Tsuchida, Tatsuya Kozaki, Hiroki Tanaka, Kyung-Sue Shin, Himanshu Kumar, and Shizuo Akira
RIG-I-like receptors (RLRs) sense virus-derived RNA or polyinosinic-polycytidylic acid (poly IC) to exert antiviral
immune responses. Akira’s group
found the mechanisms underlying the
adjuvant effects of poly IC. Poly IC wastaken up by dendritic cells (DCs), andit induced lysosomal destabilization,
which, in turn, activated an RLR-dependent signaling pathway. Upon poly IC
HMGB1 released by dying dendritic cells enhanced IFN-β production in concert with poly IC.
stimulation, cathepsin D was released
into the cytoplasm from the lysosome
number of DCs. HMGB1 released by dying cells enhanced
to interact with IPS-1, an adaptor molecule for RLRs. This inIFN-β production in concert with poly IC. Collectively, theseteraction facilitated cathepsin D cleavage of caspase 8 andfindings suggest that cathepsin D-triggered, IPS-1-dependent
the activation of the transcription factor NF-κB, resulting in
necroptosis is a mechanism that propagates the adjuvant effienhanced cytokine production. Further recruitment of the kicacy of poly IC.
nase RIP-1 to this complex initiated the necroptosis of a small

106

Takuya Uehata, Hidenori Iwasaki, Alexis Vandenbon, Kazufumi Matsushita, Eduardo Hernandez-Cuellar, Kanako Kuniyoshi,
Takashi Satoh, Takashi Mino, Yutaka Suzuki, Daron M. Standley, Tohru Tsujimura, Hiromi Rakugi, Yoshitaka Isaka, Osamu Takeuchi,
and Shizuo Akira

Although Regnase-1 (also known as Zc3h12a and MCPIP1)
inactivation leads to development of an autoimmune diseasecharacterized by T cell activation and hyperimmunoglobulinemia in mice, the mechanism of Regnase-1-mediated immune
regulation has remained unclear. Akira’s group show that
Regnase-1 is essential for preventing aberrant effector CD4+
T cell generation cell autonomously. Moreover, in T cells,
Regnase-1 regulates the mRNAs of a set of genes, including
c-Rel, Ox40, and Il2, through cleavage of their 30 UTRs. Interestingly, T cell receptor (TCR) stimulation leads to cleavageof Regnase-1 at R111 by Malt1/paracaspase, freeing T cells
from Regnase-1-mediated suppression. Furthermore, Malt1
protease activity is critical for controlling the mRNA stabilityof T cell effector genes. Collectively, these results indicate thatdynamic control of Regnase-1 expression in T cells is criticalfor controlling T cell activation.

TCR signaling leads to Regnase-1 cleavage by Malt1, facilitating T
cell activation.

107

Research Outputs

Selected Articles
Repression of the Transcription Factor Bach2 Contributes to Predisposition of IgG1 Memory B
Cells toward Plasma Cell Differentiation

Autoantibodies to IgG/HLA class II complexes are associated with rheumatoid arthritis
susceptibility

Immunity 39(1):136-47, 2013.

Proc Natl Acad Sci USA, 2014. in press.

Kohei Kometani, Rinako Nakagawa, Ryo Shinnakasu, Tomohiro Kaji, Andrei Rybouchkin, Saya Moriyama, Koji Furukawa, Haruhiko
Koseki, Toshitada Takemori, and Tomohiro Kurosaki

Hui Jin, Noriko Arase, Kouyuki Hirayasu, Masako Kohyama, Tadahiro Suenaga, Fumiji Saito, Kenji Tanimura, Sumiko Matsuoka,
Kosuke Ebina, Kenrin Shi, Noriko Toyama-Sorimachi, Shinsuke Yasuda, Tetsuya Horita, Ryosuke Hiwa, Kiyoshi Takasugi, Koichiro
Ohmura, Hideki Yoshikawa, Takashi Saito, Tatsuya Atsumi, Takehiko Sasazuki, Ichiro Katayama, Lewis L. Lanier, and Hisashi Arase

Memory B cells are essential for generating rapid and robust
secondary antibody responses. It has been thought that theunique cytoplasmic domain of IgG causes the prompt activation of antigen-experienced IgG memory B cells. Kurosaki’s
group have generated a mouse containing IgG1 B cells thathave never encountered antigen. They found that, uponchallenge, antigen-experienced IgG1 memory B cells rapidly
differentiated into plasma cells, whereas nonexperienced IgG1
B cells did not, suggesting the importance of the stimulation
history. In addition, their results suggest that repression of the
Bach2 transcription factor, which results from antigen experience, contributes to predisposition of IgG1memoryBcells todifferentiate into plasma cells.

Antigen experience induces repression of Bach2 in IgG1 memory B
cells, which in turn contributes to rapid humoral recall responses.

Strawberry notch homologue 2 regulates osteoclast fusion by enhancing the expression of DCSTAMP

Arase’s group found a unique function of HLA class II molecules: their ability to aberrantly transport cellular misfolded
proteins to the cell surface without processing to peptides.
Rheumatoid factor (RF) is an autoantibody that binds to denatured IgG or Fc fragments of IgG and is detected in 70-80%
of RA patients but also in patients with other diseases. They
report that intact IgG heavy chain (IgGH) is transported tothe cell surface by HLA class II via association with the peptide-binding groove and that IgGH/HLA class II complexes are
specifically recognized by autoantibodies in RF-positive serafrom RA patients. In contrast, autoantibodies in RF-positive
sera from non-RA individuals did not bind to IgGH/HLA class
II complexes. Of note, a strong correlation between autoantibody binding to IgG complexed with certain HLA-DR alleles
and the odds ratio for that allele's association with RA wasobserved. Their findings suggest that IgGH complexed withcertain HLA class II alleles is a target for autoantibodies in RA,
which might explain why these HLA class II alleles confer susceptibility to RA.

IgGH/HLA class II complexes are recognized by autoantibodies
in RF-positive sera from RA patients.

J Exp Med. 210(10):1947-60, 2013.
Kenta Maruyama, Satoshi Uematsu, Takeshi Kondo, Osamu Takeuchi, Mikaël M. Martino, Takumi Kawasaki, and Shizuo Akira
Osteoclasts are multinucleated cells formed by fusion ofmononuclear precursors in response to receptor activatorof nuclear factor κB (NF-κB) ligand (RANKL). Akira’s group
found that RANKL induced expression of the DExD/H helicase
family corepressor strawberry notch homologue 2 (Sbno2).
Sbno2-deficient mice exhibited increased bone mass due toimpaired osteoclast fusion. Expression of dendritic cell-specific transmembrane protein (DC-STAMP), a critical player inosteoclast fusion, was significantly attenuated, and cell fusionof Sbno2-deficient osteoclasts was rescued by DC-STAMP.
Sbno2 directly bound to T cell acute lymphocytic leukemia 1(Tal1) and attenuated its inhibition of DC-STAMP expression,leading to activation of the DC-STAMP promoter by microphthalmia-associated transcription factor (MITF). Thus, Sbno2
plays a pivotal role in bone homeostasis in vivo by fine-tuning
osteoclast fusion.

Articles, Lectures, and Awardees
FY

ArticlesLectures at
International
Meetings

Awardee(International &Domestic)

2011
2012
2013

153
179

96
99

13
15

181

127

11

Papers on high-impact journals by author(s) affiliated with IFReC

Sbno2 directly bound to Tal1 and attenuated its inhibition of
DC-STAMP expression, leading to activation of the DC-STAMP
promoter by MITF.

Nat Rev
ImmunolHigh
Impact
Total

51.7

33.1

-

1

0

0

20

0

1

0

1

21

1

1

1

3

16

Nat
Nat
New Eng
Nat MedScience
Immunol
Neurosci
J Med

FY

Cell

Immunity

IF*

32.4

24.2

14.8

36.0

25.7

25.4

14.2

31.4

2011

1

9

3

0

4

2

0

2012

1

9

3

3

2

1

2013

2

3

1

2

2

0

JEM Nature

Data from WEB OF SCIENCE™
*The impact factor means the frequency with which the “average article” in a journal has been cited in a given period of time.

108

109

Research Outputs

Publications
1

Akira, S.; Misawa, T.; Satoh, T.; Saitoh, T.. Macrophages control innate
inflammation. Diabetes Obesity & Metabolism 15:10-18, 2013.

2

Aksoy, Irene; Jauch, Ralf; Chen, Jiaxuan; Dyla, Mateusz; Divakar,
Ushashree; Bogu, Gireesh K.; Teo, Roy; Ng, Calista Keow Leng; Herath,
Wishva; Sun Lili; Hutchins, Andrew P.; Robson, Paul; Kolatkar, PrasannaR.; Stanton, Lawrence W.. Oct4 switches partnering from Sox2 to
Sox17 to reinterpret the enhancer code and specify endoderm. Embo
Journal 32:938-953, 2013.

3

4

Atif, S. M.; Uematsu, S.; Ak ira, S.; M cS orley, S. J.. CD103CD11b+dendritic cells regulate the sensitivity of CD4 T-cell responses
to bacterial flagellin. Mucosal Immunology 7:68-77, 2014.

5

Atsumi, Toru; Singh, Rajeev; Sabharwal, Lavannya; Bando, Hidenori;Meng, Jie; Arima, Yasunobu; Yamada, Moe; Harada, Masaya; Jiang,
Jing-Jing; Kamimura, Daisuke; Ogura, Hideki; Hirano, Toshio;
Murakami, Masaaki. Inflammation Amplifier, a New Paradigm inCancer Biology. Cancer Research 74:8-14, 2014.

6

Banerji, Christopher R. S.; Miranda-Saavedra, Diego; Severini, Simone;
Widschwendter, Martin; Enver, Tariq; Zhou, Joseph X.; Teschendorff,
Andrew E.. Cellular network entropy as the energy potential in
Waddington's differentiation landscape. Scientific Reports 3:3039,
2013.

7

Bussieres-Marmen, Stephanie; Hutchins, Andrew P.; Schirbel, Anja;
Rebert, Nancy; Tiganis, Tony; Fiocchi, Claudio; Miranda-Saavedra,
Diego; Tremblay, Michel L.. Characterization of PTPN2 and its use as abiomarker. Methods 65:239-246, 2014.

8

Chen, James K.; Kikuchi, Kazuya. Emerging technologies in molecularimaging: new windows into biology. Current Opinion In Chemical
Biology 17:635-636, 2013.

9

Chiyonobu, Tomohiro; Inoue, Norimitsu; Morimoto, Masafumi;
Kinoshita, Taroh; Murakami, Yoshiko. Glycosylphosphatidylinositol(GPI) anchor deficiency caused by mutations in PIGW is associatedwith West syndrome and hyperphosphatasia with mental retardation
syndrome. Journal of Medical Genetics 51:203-207, 2014.

10

Chow, Ho Ming; Horovitz, Silvina G.; Carr, Walter S.; Picchioni, Dante;
Coddington, Nate; Fukunaga, Masaki; Xu, Yisheng; Balkin, Thomas J.;
Duyn, Jeff H.; Braun, Allen R.. Rhythmic alternating patterns of brain
activity distinguish rapid eye movement sleep from other states ofconsciousness. Proceedings of National Academy of Sciences USA
110:10300-10305, 2013.

11

Coban, Cevayir; Kobiyama, Kouji; Jounai, Nao; Tozuka, Miyuki; Ishii,
Ken J.. DNA vaccines A simple DNA sensing matter?. Human Vaccines& Immunotherapeutics 9:2216-2221, 2013.

12

Cui, Guangwei; Hara, Takahiro; Simmons, Szandor; Wagatsuma,
Keisuke; Abe, Akifumi; Miyachi, Hitoshi; Kitano, Satsuki; Ishii, Masaru;
Tani-ichi, Shizue; Ikuta, Koichi. Characterization of the IL-15 niche inprimary and secondary lymphoid organs in vivo. Proceedings of The
National Academy of Sciences, USA 111:1915-1920, 2014.

13

Deretic, Vojo; Saitoh, Tatsuya; Akira, Shizuo. Autophagy in infection,
inflammation and immunity. Nature Reviews Immunology 13:722737, 2013.

14

Diez, Diego; Hutchins, Andrew Paul; Miranda-Saavedra, Diego.
Systematic identification of transcriptional regulatory modules fromprotein-protein interaction networks. Nucleic Acids Research 42:e6,
2014.

15

Dorner, Marcus; Horwitz, Joshua A.; Donovan, Bridget M.; Labitt,
Rachael N.; Budell, William C.; Friling, Tamar; Vogt, Alexander;
Catanese, Maria Teresa; Satoh, Takashi; Kawai, Taro; Akira, Shizuo; Law,
Mansun; Rice, Charles M.; Ploss, Alexander. Completion of the entire
hepatitis C virus life cycle in genetically humanized mice. Nature
501:237-241, 2013.

16

17
110

Atarashi, Koji; Tanoue, Takeshi; Oshima, Kenshiro; Suda, Wataru;
Nagano, Yuji; Nishikawa, Hiroyoshi; Fukuda, Shinji; Saito, Takuro;
Narushima, Seiko; Hase, Koji; Kim, Sangwan; Fritz, Joelle V.; Wilmes,
Paul; Ueha, Satoshi; Matsushima, Kouji; Ohno, Hiroshi; Olle, Bernat;
Sakaguchi, Shimon; Taniguchi, Tadatsugu; Morita, Hidetoshi; Hattori,
Masahira; Honda, Kenya. T-reg induction by a rationally selected
mixture of Clostridia strains from the human microbiota. Nature
500:232-236, 2013.

18

Forrest, Alistair R. R. et al. A promoter-level mammalian expression
atlas. Nature 507:462-470, 2014.

19

Fujita, Naonobu; Morita, Eiji; Itoh, Takashi; Tanaka, Atsushi; Nakaoka,
Megumi; Osada, Yuki; Umemoto, Tetsuo; Saitoh, Tatsuya; Nakatogawa,
Hitoshi; Kobayashi, Shouhei; Haraguchi, Tokuko; Guan, Jun-Lin; Iwai,
Kazuhiro; Tokunaga, Fuminori; Saito, Kazunobu; Ishibashi, Koutaro;
Akira, Shizuo; Fukuda, Mitsunori; Noda, Takeshi; Yoshimori, Tamotsu.
Recruitment of the autophagic machinery to endosomes during
infection is mediated by ubiquitin. Journal of Cell Biology 203:115128, 2013.

20

Fujita, Toshitsugu; Asano, Yoshinori; Ohtsuka, Junko; Takada, Yoko;
Saito, Kazunobu; Ohki, Rieko; Fujii, Hodaka. Identification of telomereassociated molecules by engineered DNA-binding moleculemediated chromatin immunoprecipitation (enChIP). Scientific
Reports 3:UNSP 3171, 2013.

21

Fujiwara, Nagatoshi; Porcelli, Steven A.; Naka, Takashi; Yano, Ikuya;
Maeda, Shinji; Kuwata, Hirotaka; Akira, Shizuo; Uematsu, Satoshi;
Takii, Takemasa; Ogura, Hisashi; Kobayashi, Kazuo. Bacterial
sphingophospholipids containing non-hydroxy fatty acid activate
murine macrophages via Toll-like receptor 4 and stimulate bacterial
clearance. Biochimica Et Biophysica Acta-Molecular And Cell Biology
Of Lipids 1831:1177-1184, 2013.

22

Fukasaka, Masahiro; Ori, Daisuke; Kawagoe, Tatsukata; Uematsu,
Satoshi; Maruyama, Kenta; Okazaki, Toshihiko; Kozaki, Tatsuya;
Imamura, Tomoko; Tartey, Sarang; Mino, Takashi; Satoh, Takashi; Akira,Shizuo; Takeuchi, Osamu. Critical Role of AZI2 in GM-CSF-Induced
Dendritic Cell Differentiation. Journal of Immunology 190:5702-5711,
2013.

23

Fukui, Michiaki; Ushigome, Emi; Tanaka, Muhei; Hamaguchi,
Masahide; Tanaka, Toru; Atsuta, Haruhiko; Ohnishi, Masayoshi;
Oda, Yohei; Hasegawa, Goji; Nakamura, Naoto. Could home arterial
stiffness index be a novel marker for arterial stiffness in patients with
type 2 diabetes?. Hypertension Research 36:645-649, 2013.

24

Goritzka, M.; Durant, L. R.; Pereira, C. R.; Makris, S.; Kausar, F.; Kumagai,Y.; Akira, S.; Johansson, C.. Alveolar macrophages modulate early
immune responses to Respiratory Syncytial Virus (RSV) infection
through MAVS signalling and type I interferons. Immunology 140:8787, 2013.

25

Haldar, Malay; Kohyama, Masako; So, Alex Yick-Lun; Wumesh, K. C.;
Wu, Xiaodi; Briseno, Carlos G.; Satpathy, Ansuman T.; Kretzer, Nicole
M.; Arase, Hisashi; Rajasekaran, Namakkal S.; Wang, Li; Egawa, Takeshi;
Igarashi, Kazuhiko; Baltimore, David; Murphy, Theresa L.; Murphy,
Kenneth M.. Heme-Mediated SPI-C Induction Promotes Monocyte
Differentiation into Iron-Recycling Macrophages. Cell 156:1223-1234,
2014.

26

Halder, Sebok K.; Matsunaga, Hayato; Ishii, Ken J.; Akira, Shizuo;
Miyake, Kensuke; Ueda, Hiroshi. Retinal cell type-specific preventionof ischemia-induced damages by LPS-TLR4 signaling throughmicroglia. Journal of Neurochemistry 126:243-260, 2013.

27

Hamada, Michito; Nakamura, Megumi; Mai Thi Nhu Tran; Moriguchi,
Takashi; Hong, Cynthia; Ohsumi, Takayuki; Tra Thi Huong Dinh;
Kusakabe, Manabu; Hattori, Motochika; Katsumata, Tokio; Arai, Satoko;
Nakashima, Katsuhiko; Kudo, Takashi; Kuroda, Etsushi; Wu, ChienHui; Kao, Pei-Han; Sakai, Masaharu; Shimano, Hitoshi; Miyazaki, Toru;
Tontonoz, Peter; Takahashi, Satoru. MafB promotes atherosclerosis by
inhibiting foam-cell apoptosis. Nature Communications 5:3147, 2014.

28

29

Hansen, Lars; Tawamie, Hasan; Murakami, Yoshiko; Mang, Yuan;
Rehman, Shoaib Ur; Buchert, Rebecca; Schaffer, Stefanie; Muhammad,
Safia; Bak, Mads; Noethen, Markus M.; Bennett, Eric P.; Maeda, Yusuke;
Aigner, Michael; Reis, Andre; Kinoshita, Taroh; Tommerup, Niels;
Baig, Shahid Mahmood; Abou Jamra, Rami. Hypomorphic Mutationsin PGAP2, Encoding a GPI-Anchor-Remodeling Protein, Cause
Autosomal-Recessive Intellectual Disability. American Journal of
Human Genetics 92:575-583, 2013.
Hatzihristidis, Teri; Liu, Shaq; Pryszcz, Leszek; Hutchins, Andrew P.;
Gabaldon, Toni; Tremblay, Michel L.; Miranda-Saavedra, Diego. PTPcentral: A comprehensive resource of protein tyrosine phosphatases
in eukaryotic genomes. Methods 65:156-164, 2014.

Enokizono, Yoshiaki; Kumeta, Hiroyuki; Funami, Kenji; Horiuchi,
Masataka; Sarmiento, Joy; Yamashita, Kazuo; Standley, Daron M.;
Matsumoto, Misako; Seya, Tsukasa; Inagaki, Fuyuhiko. Structures andinterface mapping of the TIR domain-containing adaptor molecules
involved in interferon signaling. Proceedings of National Academy of
Sciences USA 110:19908-19913, 2013.

30

H i r a t a , Te t s u y a ; F u j i t a , M o r i h i s a ; K a n z a w a , N o r i y u k i ;M u r a k a m i , Yo s h i k o ; M a e d a , Yu s u k e ; K i n o s h i t a , Ta r o h .
Glycosylphosphatidylinositol mannosyltransferase II is the ratelimiting enzyme in glycosylphosphatidylinositol biosynthesis
under limited dolichol-phosphate mannose availability. Journal of
Biochemistry 154:257-264, 2013.

Feldhammer, Matthew; Uetani, Noriko; Miranda-Saavedra, Diego;
Tremblay, Michel L.. PTP1B: A simple enzyme for a complex world.
Critical Reviews In Biochemistry And Molecular Biology 48:430-445,
2013.

31

Hirayama, Michiko; Nishikawa, Hiroyoshi; Nagata, Yasuhiro; Tsuji,
Takemasa; Kato, Takuma; Kageyama, Shinichi; Ueda, Shugo;
Sugiyama, Daisuke; Hori, Sahoko; Sakaguchi, Shimon; Ritter, Gerd;
Old, Lloyd J.; Gnjatic, Sacha; Shiku, Hiroshi. Overcoming regulatory
T-cell suppression by a lyophilized preparation of Streptococcus
pyogenes. European Journal of Immunology 43:989-1000, 2013.

32

Hori, Yuichiro; Kikuchi, Kazuya. Protein labeling with fluorogenic
probes for no-wash live-cell imaging of proteins. Current Opinion In
Chemical Biology 17:644-650, 2013.

33

Hori, Yuichiro; Norinobu, Tomoya; Sato, Motoki; Arita, Kyohei;
Shirakawa, Masahiro; Kikuchi, Kazuya. Development of Fluorogenic
Probes for Quick No-Wash Live-Cell Imaging of Intracellular Proteins.
Journal of The American Chemical Society 135:12360-12365, 2013.

34

Hoshii, Takayuki; Kasada, Atsuo; Hatakeyama, Tomoki; Ohtani,
Masashi; Tadokoro, Yuko; Naka, Kazuhito; Ikenoue, Tsuneo; Ikawa,
Tomokatsu; Kawamoto, Hiroshi; Fehling, Hans Joerg; Araki, Kimi;
Yamamura, Ken-ichi; Matsuda, Satoshi; Hirao, Atsushi. Loss of mTOR
complex 1 induces developmental blockage in early T-lymphopoiesis
and eradicates T-cell acute lymphoblastic leukemia cells. Proceedingsof The National Academy of Sciences, USA 111:3805-3810, 2014.

35

Hosokawa, Hiroyuki; Tanaka, Tomoaki; Kato, Miki; Shinoda, Kenta;
Tohyama, Hiroyuki; Hanazawa, Asami; Tamaki, Yuuki; Hirahara, Kiyoshi;
Yagi, Ryoji; Sakikawa, Ikue; Morita, Atsushi; Nagira, Morio; Poyurovsky,
Masha V.; Suzuki, Yutaka; Motohashi, Shinichiro; Nakayama, Toshinori.
Gata3/Ruvbl2 complex regulates T helper 2 cell proliferation via
repression of Cdkn2c expression. Proceedings of National Academyof Sciences USA 110:18626-18631, 2013.

36

Howard, Malcolm F.; Murakami, Yoshiko; Pagnamenta, Alistair T.;
Daumer-Haas, Cornelia; Fischer, Bjoern; Hecht, Jochen; Keays, David
A.; Knight, Samantha J. L.; Koelsch, Uwe; Krueger, Ulrike; Leiz, Steffen;
Maeda, Yusuke; Mitchell, Daphne; Mundlos, Stefan; Phillips, John
A., III; Robinson, Peter N.; Kini, Usha; Taylor, Jenny C.; Horn, Denise;
Kinoshita, Taroh; Krawitz, Peter M.. Mutations in PGAP3 Impair GPIAnchor Maturation, Causing a Subtype of Hyperphosphatasia with
Mental Retardation. American Journal of Human Genetics 94:278287, 2014.

37

Hutchins, Andrew P.; Diez, Diego; Miranda-Saavedra, Diego. The IL-10/
STAT3-mediated anti-inflammatory response: recent developments
and future challenges. Briefings In Functional Genomics 12:489-498,
2013.

38

Hutchins, Andrew Paul; Liu, Shaq; Diez, Diego; Miranda-Saavedra,
Diego. The Repertoires of Ubiquitinating and Deubiquitinating
Enzymes in Eukaryotic Genomes. Molecular Biology And Evolution
30:1172-1187, 2013.

39

Ichimura, Taro; Chiu, Liang-da; Fujita, Katsumasa; Kawata, Satoshi;
Watanabe, Tomonobu M.; Yanagida, Toshio; Fujita, Hideaki. Visualizing
Cell State Transition Using Raman Spectroscopy. Plos One 9:e84478,
2014.

40

Imai, Takahiko; Koyanagi, Naoto; Ogawa, Ryo; Shindo, Keiko; Suenaga,
Tadahiro; Sato, Ayuko; Arii, Jun; Kato, Akihisa; Kiyono, Hiroshi;
Arase, Hisashi; Kawaguchi, Yasushi. Us3 Kinase Encoded by Herpes
Simplex Virus 1 Mediates Downregulation of Cell Surface Major
Histocompatibility Complex Class I and Evasion of CD8(+) T Cells.
PLOS ONE 8:e72050, 2013.

41

Inoue, Takahiro; Iijima, Hideki; Arimitsu, Junsuke; Hagihara,
Keisuke; Kawai, Shoichiro; Shiraishi, Eri; Hiyama, Satoshi; Mukai,
Akira; Shinzaki, Shinichiro; Nishida, Tsutomu; Ogata, Atsushi; Tsujii,
Masahiko; Takehara, Tetsuo. Amelioration of Small Bowel Injury by
Switching from Nonselective Nonsteroidal Anti-Inflammatory Drugs
to Celecoxib in Rheumatoid Arthritis Patients: A Pilot Study. Digestion
89:124-132, 2014.

47

Ito, Koyu; Morimoto, Junko; Kihara, Akio; Matsui, Yutaka; Kurotaki,
Daisuke; Kanayama, Masashi; Simmons, Szandor; Ishii, Masaru;
Sheppard, Dean; Takaoka, Akinori; Uede, Toshimitsu. Integrin alpha9 on lymphatic endothelial cells regulates lymphocyte egress.
Proceedings of The National Academy of Sciences, USA 111:30803085, 2014.

48

Iyer, Smita S.; Latner, Donald R.; Zilliox, Michael J.; McCausland,
Megan; Akondy, Rama S.; Penaloza-MacMaster, Pablo; Hale, Jeffrey
Scott; Ye, Lilin; Mohammed, Ata-Ur-Rasheed; Yamaguchi, Tomoyuki;
Sakaguchi, Shimon; Amara, Rama R.; Ahmed, Rafi. Identification ofnovel markers for mouse CD4(+) T follicular helper cells. European
Journal of Immunology 43:3219-3232, 2013.

49

Jiang, Yan; Arase, Noriko; Kohyama, Masako; Hirayasu, Kouyuki;
Suenaga, Tadahiro; Jin, Hui; Matsumoto, Maki; Shida, Kyoko; Lanier,
Lewis L.; Saito, Takashi; Arase, Hisashi. Transport of misfolded
endoplasmic reticulum proteins to the cell surface by MHC class II
molecules. International Immunology 25:235-246, 2013.

50

Jin, Hui; Arase, Noriko; Hirayasu, Kouyuki; Kohyama, Masako; Suenaga,
Tadahiro; Saito, Fumiji; Tanimura, Kenji; Matsuoka, Sumiko; Ebina,
Kosuke; Shi, Kenrin; Toyama-Sorimachi, Noriko; Yasuda, Shinsuke;
Horita, Tetsuya; Hiwa, Ryosuke; Takasugi, Kiyoshi; Ohmura, Koichiro;
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102

Mizukami, Shin; Hori, Yuichiro; Kikuchi, Kazuya. Small-MoleculeBased Protein-Labeling Technology in Live Cell Studies: Probe-Design
Concepts and Applications. Accounts of Chemical Research 47:247256, 2014.

103

Moretti, Rocco. et al. Community-wide evaluation of methods forpredicting the effect of mutations on protein-protein interactions.
Proteins-Structure Function And Bioinformatics 81:1980-1987, 2013.

104

Mori, Yuki; Murakami, Masaaki; Arima, Yasunobu; Zhu, Dasong;
Terayama, Yasuo; Komai, Yutaka; Nakatsuji, Yuji; Kamimura, Daisuke;
Yoshioka, Yoshichika. Early pathological alterations of lower lumbar
cords detected by ultrahigh-field MRI in a mouse multiple sclerosis
model. International Immunology 26:93-101, 2014.

105

Morimoto, Yasuo; Oyabu, Takako; Horie, Masanori; Kambara,
Tatsunori; Izumi, Hiroto; Kuroda, Etsushi; Creutzenberg, Otto;
Bellmann, Bernd; Pohlmann, Gerhard; Schuchardt, Sven; Hansen,
Tanja; Ernst, Heinrich. Pulmonary toxicity of printer toner following
inhalation and intratracheal instillation. Inhalation Toxicology 25:679690, 2013.

106

Mosiewicz, Katarzyna A.; Kolb, Laura; van der Vlies, Andre J.; Martino,
Mikael M.; Lienemann, Philipp S.; Hubbell, Jeffrey A.; Ehrbar, Martin;
Lutolf, Matthias P.. In situ cell manipulation through enzymatichydrogel photopatterning. Nature Materials 12:1071-1077, 2013.

113

Research Outputs

121

Ohi, Kazutaka; Hashimoto, Ryota; Yamamori, Hidenaga; Yasuda,
Yuka; Fujimoto, Michiko; Umeda-Yano, Satomi; Fukunaga, Masaki;
Watanabe, Yoshiyuki; Iwase, Masao; Kazui, Hiroaki; Takeda, Masatoshi.
The impact of the genome-wide supported variant in the cyclin M2
gene on gray matter morphology in schizophrenia. Behavioral andBrain Functions 9:40, 2013.

122

Ohta, Katsuya; Haraguchi, Naotsugu; Kano, Yoshihiro; Kagawa,Yoshinori; Konno, Masamitsu; Nishikawa, Shimpei; Hamabe, Atsushi;
Hasegawa, Shinichiro; Ogawa, Hisataka; Fukusumi, Takahito;
Uemura, Mamoru; Nishimura, Junichi; Hata, Taishi; Takemasa, Ichiro;
Mizushima, Tsunekazu; Noguchi, Yuko; Ozaki, Miyuki; Kudo, Toshihiro;
Sakai, Daisuke; Satoh, Taroh; Fukami, Miwa; Ishii, Masaru; Yamamoto,Hirofumi; Doki, Yuichiro; Mori, Masaki; Ishii, Hideshi. Depletion of
JARID1B induces cellular senescence in human colorectal cancer.
International Journal of Oncology 42:1212-1218, 2013.

123

124

Ono, Chikako; Ninomiya, Akinori; Yamamoto, Satomi; Abe, Takayuki;
Wen, Xiauyu; Fukuhara, Takasuke; Sasai, Miwa; Yamamoto, Masahiro;
Saitoh, Tatsuya; Satoh, Takashi; Kawai, Taro; Ishii, Ken J.; Akira, Shizuo;
Okamoto, Toru; Matsuura, Yoshiharu. Innate Immune Response
Induced by Baculovirus Attenuates Transgene Expression in
Mammalian Cells. Journal of Virology 88:2157-2167, 2014.

125

Oo, Ye Htun; Sakaguchi, Shimon. Regulatory T-cell directed therapies
in liver diseases. Journal of Hepatology 59:1127-1134, 2013.

126

Ori, Daisuke; Kato, Hiroki; Sanjo, Hideki; Tartey, Sarang; Mino, Takashi;
Akira, Shizuo; Takeuchi, Osamu. Essential Roles of K63-Linked
Polyubiquitin-Binding Proteins TAB2 and TAB3 in B Cell Activation via
MAPKs. Journal of Immunology 190:4037-4045, 2013.

127

Oyabu, Takako; Morimoto, Yasuo; Hirohashi, Masami; Horie,
Masanori; Kambara, Tatsunori; Lee, Byeong Woo; Hashiba, Masayoshi;
Mizuguchi, Yohei; Myojo, Toshihiko; Kuroda, Etsushi. Dose-dependent
pulmonary response of well-dispersed titanium dioxide nanoparticles
following intratracheal instillation. Journal of Nanoparticle Research
15:UNSP 1600, 2013.

128

129

130

114

Ohta, Tomokazu; Koshi, Katsuo; Ushizawa, Koichi; Hosoe, Misa;
Takahashi, Toru; Yamaguchi, Takahiro; Kizaki, Keiichiro; Hashizume,
Kazuyoshi. Dynamics of CD3(+) T-cell Distribution Throughout the
Estrous Cycle and Gestation in the Bovine Endometrium. Journal of
Reproduction And Development 59:507-511, 2013.

Palacpac, Nirianne Marie Q.; Ntege, Edward; Yeka, Adoke; Balikagala,
Betty; Suzuki, Nahoko; Shirai, Hiroki; Yagi, Masanori; Ito, Kazuya;
Fukushima, Wakaba; Hirota, Yoshio; Nsereko, Christopher; Okada,
Takuya; Kanoi, Bernard N.; Tetsutani, Kohhei; Arisue, Nobuko; Itagaki,
Sawako; Tougan, Takahiro; Ishii, Ken J.; Ueda, Shigeharu; Egwang,Thomas G.; Horii, Toshihiro. Phase 1b Randomized Trial and FollowUp Study in Uganda of the Blood-Stage Malaria Vaccine Candidate
BK-SE36. PLOS ONE 8:e64073, 2013.
Patel, Khushbu K.; Miyoshi, Hiroyuki; Beatty, Wandy L.; Head, RichardD.; Malvin, Nicole P.; Cadwell, Ken; Guan, Jun-Lin; Saitoh, Tatsuya;
Akira, Shizuo; Seglen, Per O.; Dinauer, Mary C.; Virgin, Herbert W.;
Stappenbeck, Thaddeus S.. Autophagy proteins control goblet cellfunction by potentiating reactive oxygen species production. Embo
Journal 32:3130-3144, 2013.
Patil, Ashwini; Kumagai, Yutaro; Liang, Kuo-ching; Suzuki, Yutaka;
Nakai, Kenta. Linking Transcriptional Changes over Time in Stimulated
Dendritic Cells to Identify Gene Networks Activated during the Innate
Immune Response. Plos Computational Biology 9:e1003323, 2013.

131

Pavillon, Nicolas; Bando, Kazuki; Fujita, Katsumasa; Smith, Nicholas I..
Feature-based recognition of surface-enhanced Raman spectra forbiological targets. Journal of Biophotonics 6:587-597, 2013.

132

Pike, K. -A.; Hutchins, A. P.; Tremblay, M. L.; Miranda-Saavedra, D..
Systems biology approaches to dissect and manipulate the IL-10/
STAT3 anti-inflammatory response. Immunology 140:36-36, 2013.

133

Pissuwan, D.; Hobro, A. J.; Pavillon, N.; Smith, N. I.. Distribution of label
free cationic polymer-coated gold nanorods in live macrophage cells
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134

Pissuwan, Dakrong; Kumagai, Yutaro; Smith, Nicholas I.. Effect ofSurface-Modified Gold Nanorods on the Inflammatory Cytokine
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135

Rattanapak, Teerawan; Birchall, James C.; Young, Katherine; Kubo,
Atsuko; Fujimori, Sayumi; Ishii, Masaru; Hook, Sarah. Dynamic
Visualization of Dendritic Cell-Antigen Interactions in the Skin
Following Transcutaneous Immunization. Plos One 9:e89503, 2014.

136

Rattanapak, Teerawan; Birchall, James; Young, Katherine; Ishii,
Masaru; Meglinski, Igor; Rades, Thomas; Hook, Sarah. Transcutaneous
immunization using microneedles and cubosomes: Mechanistic
investigations using Optical Coherence Tomography and Two-Photon
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151

Sultana, Tahmina; Takagi, Hiroaki; Morimatsu, Miki; Teramoto, Hiroshi;
Li, Chun-Biu; Sako, Yasushi; Komatsuzaki, Tamiki. Non-Markovian
properties and multiscale hidden Markovian network buried in single
molecule time series. Journal of Chemical Physics 139:245101, 2013.

137

Sahoo, Bikash R.; Maharana, Jitendra; Bhoi, Gopal K.; Lenka, Santosh
K.; Patra, Mahesh C.; Dikhit, Manas R.; Dubey, Praveen K.; Pradhan,
Sukanta K.; Behera, Bijay K.. A conformational analysis of mouse
Nalp3 domain structures by molecular dynamics simulations, andbinding site analysis. Molecular Biosystems 10:1104-1116, 2014.

152

Tachino, Sho; Fujiki, Fumihiro; Oka, Yoshihiro; Tsuboi, Akihiro;
Morimoto, Soyoko; Lin, Yu-Hung; Tamanaka, Taichi; Kondo, Kenta;
Nakajima, Hiroko; Nishida, Sumiyuki; Hosen, Naoki; Oji, Yusuke;
Kumanogoh, Atsushi; Sugiyama, Haruo. Functional human Th17
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138

Saito, H.; Ogasawara, K.; Nishimoto, H.; Yoshioka, Y.; Murakami, T.;
Fujiwara, S.; Sasaki, M.; Kobayashi, M.; Yoshida, K.; Kubo, Y.; Beppu, T.;
Ogawa, A.. Postoperative Changes in Cerebral Metabolites Associatedwith Cognitive Improvement and Impairment after Carotid
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153

Tada, Satoru; Yasui, Teruhito; Nakatsuji, Yuji; Okuno, Tatsusada; Koda,
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154

Tanaka, Toshio. Can IL-6 blockade rectify imbalance between Tregs
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155

Tanaka, Toshio; Narazaki, Masashi; Ogata, Atsushi; Kishimoto,Tadamitsu. A new era for the treatment of inflammatory autoimmune
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139

Sakaguchi, Shimon; Vignali, Dario A. A.; Rudensky, Alexander Y.;
Niec, Rachel E.; Waldmann, Herman. The plasticity and stability of
regulatory T cells. Nature Reviews Immunology 13:461-467, 2013.

140

Sandig, Hilary; Jobbings, Catherine E.; Roldan, Nestor Gonzalez;
Whittingham-Dowd, Jayde K.; Orinska, Zane; Takeuchi, Osamu; Akira,Shizuo; Bulfone-Paus, Silvia. IL-33 causes selective mast cell tolerance
to bacterial cell wall products by inducing IRAK1 degradation.
European Journal of Immunology 43:979-988, 2013.

141

Sasai, Miwa; Yamamoto, Masahiro. Pathogen Recognition Receptors:
Ligands and Signaling Pathways by Toll-Like Receptors. International
Reviews of Immunology 32:116-133, 2013.

142

Sasaki, Yoshiteru; Sano, Soichi; Nakahara, Masaki; Murata, Shigeo;
Kometani, Kohei; Aiba, Yuichi; Sakamoto, Shinji; Watanabe, Yoshihiro;
Tanaka, Keiji; Kurosaki, Tomohiro; Iwai, Kazuhiro. Defective immune
responses in mice lacking LUBAC-mediated linear ubiquitination in B
cells. Embo Journal 32:2463-2476, 2013.

143

Sato, S.; Kaneto, S.; Shibata, N.; Takahashi, Y.; Okura, H.; Yuki, Y.;
Kunisawa, J.; Kiyono, H.. Transcription factor Spi-B-dependent and-independent pathways for the development of Peyer's patch M cells.
Mucosal Immunology 6:838-846, 2013.

144

Sawada, Kazuhiko; Saito, Shigeyoshi; Horiuchi-Hirose, Miwa; Mori,
Yuki; Yoshioka, Yoshichika; Murase, Kenya. Dose-related cerebellar
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145

S e o n g , J i h y o u n ; Wa n g , Ye t a o ; K i n o s h i t a , Ta r o h ; M a e d a ,Yusuke. Implications of lipid moiety in oligomerization andimmunoreactivities of GPI-anchored proteins. Journal of Lipid
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146

Shimada, Yosuke; Kinoshita, Makoto; Harada, Kazuo; Mizutani,
Masafumi; Masahata, Kazunori; Kayama, Hisako; Takeda, Kiyoshi.
Commensal Bacteria-Dependent Indole Production Enhances
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147

Shimizu, Kanako; Asakura, Miki; Shinga, Jun; Sato, Yusuke; Kitahara,
Shuji; Hoshino, Katsuaki; Kaisho, Tsuneyasu; Schoenberger, Stephen P.;
Ezaki, Taichi; Fujii, Shin-ichiro. Invariant NKT Cells Induce Plasmacytoid
Dendritic Cell (DC) Cross-Talk with Conventional DCs for Efficient
Memory CD8(+) T Cell Induction. Journal of Immunology 190:56095619, 2013.

148

149

150

Shimozawa, Togo; Yamagata, Kazuo; Kondo, Takefumi; Hayashi,
Shigeo; Shitamukai, Atsunori; Konno, Daijiro; Matsuzaki, Fumio;
Takayama, Jun; Onami, Shuichi; Nakayama, Hiroshi; Kosugi, Yasuhito;
Watanabe, Tomonobu M.; Fujita, Katsumasa; Mimori-Kiyosue, Yuko.
Improving spinning disk confocal microscopy by preventing pinholecross-talk for intravital imaging (vol 110, pg 3399, 2013). Proceedingsof National Academy of Sciences USA 110:6240-6240, 2013.
Sugita, Kazunari; Ikenouchi-Sugita, Atsuko; Nakayama, Yasuko;
Yoshioka, Haruna; Nomura, Takashi; Sakabe, Jun-ichi; Nakahigashi,
Kyoko; Kuroda, Etsushi; Uematsu, Satoshi; Nakamura, Jun; Akira,Shizuo; Nakamura, Motonobu; Narumiya, Shuh; Miyachi, Yoshiki;
Tokura, Yoshiki; Kabashima, Kenji. Prostaglandin E-2 is critical for thedevelopment of niacin-deficiency-induced photosensitivity via ROS
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Sugiyama, Daisuke; Nishikawa, Hiroyoshi; Maeda, Yuka; Nishioka,
Megumi; Tanemura, Atsushi; Katayama, Ichiro; Ezoe, Sachiko;
Kanakura, Yuzuru; Sato, Eiichi; Fukumori, Yasuo; Karbach, Julia;
Jaeger, Elke; Sakaguchi, Shimon. Anti-CCR4 mAb selectively depletes
effector-type FoxP3(+)CD4(+) regulatory T cells, evoking antitumor
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156

Tanaka, Toshio; Takahashi, Ryo. Flavonoids and Asthma. Nutrients
5:2128-2143, 2013.

157

Tanaka, Yuki; Suenaga, Tadahiro; Matsumoto, Misako; Seya, Tsukasa;
Arase, Hisashi. Herpesvirus 6 Glycoproteins B (gB), gH, gL, and gQ Are
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87:10900-10903, 2013.

158

Tetsumoto, Satoshi; Takeda, Yoshito; Imai, Hirohiko; Kimura, Atsuomi;
Jin, Yingji; Nakanishi, Kaori; Maeda, Yohei; Kuhara, Hanako; Tsujino,
Kazuyuki; Iwasaki, Takeo; Shigeta, Hiroshi; Kondo, Yasushi; Ito, Mari;
Minami, Toshiyuki; Hirata, Haruhiko; Takahashi, Ryo; Kohmo, Satoshi;
Nagatomo, Izumi; Inoue, Koji; Kida, Hiroshi; Kijima, Takashi; Tachibana,
Isao; Maeda, Norikazu; Funahashi, Toru; Shimomura, Iichiro;
Fujiwara, Hideaki; Kumanogoh, Atsushi. Validation of Noninvasive
Morphological and Diffusion Imaging in Mouse Emphysema by
Micro-Computed Tomography and Hyperpolarized Xe-129 Magnetic
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159

Tsukumo, Shin-ichi; Unno, Midori; Muto, Akihiko; Takeuchi, Arata;
Kometani, Kohei; Kurosaki, Tomohiro; Igarashi, Kazuhiko; Saito,
Takashi. Bach2 maintains T cells in a naive state by suppressing
effector memory-related genes. Proceedings of National Academy of
Sciences USA 110:10735-10740, 2013.

160

Uehata, Takuya; Iwasaki, Hidenori; Vandenbon, Alexis; Matsushita,
Kazufumi; Hernandez-Cuellar, Eduardo; Kuniyoshi, Kanako; Satoh,
Takashi; Mino, Takashi; Suzuki, Yutaka; Standley, Daron M.; Tsujimura,
Tohru; Rakugi, Hiromi; Isaka, Yoshitaka; Takeuchi, Osamu; Akira,Shizuo. Malt1-Induced Cleavage of Regnase-1 in CD4(+) Helper T
Cells Regulates Immune Activation. Cell 153:1036-1049, 2013.

161

Ueno, Masaki; Fujita, Yuki; Tanaka, Tatsuhide; Nakamura, Yuka; Kikuta,
Junichi; Ishii, Masaru; Yamashita, Toshihide. Layer V cortical neurons
require microglial support for survival during postnatal development.
Nature Neuroscience 16:543-551, 2013.

162

Umemoto, Eiji; Takeda, Akira; Jin, Soojung; Luo, Zhijuan; Nakahogi,
Naoki; Hayasaka, Haruko; Lee, Chun Man; Tanaka, Toshiyuki; Miyasaka,
Masayuki. Dynamic Changes in Endothelial Cell Adhesion Molecule
Nepmucin/CD300LG Expression under Physiological and Pathological
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163

Ushigome, Emi; Fukui, Michiaki; Hamaguchi, Masahide; Matsumoto,
Shinobu; Mineoka, Yusuke; Nakanishi, Naoko; Senmaru, Takafumi;
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pulse pressure is associated more strongly with elevated albuminuria
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164

Ushigome, Emi; Fukui, Michiaki; Hamaguchi, Masahide; Tanaka, Toru;
Atsuta, Haruhiko; Ohnishi, Masayoshi; Oda, Yohei; Yamazaki, Masahiro;
Hasegawa, Goji; Nakamura, Naoto. Beneficial effect of calcium
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4:399-404, 2013.

165

Wang, Yetao; Murakami, Yoshiko; Yasui, Teruhito; Wakana, Shigeharu;
Kikutani, Hitoshi; Kinoshita, Taroh; Maeda, Yusuke. Significanceof Glycosylphosphatidylinositol-anchored Protein Enrichment in
Lipid Rafts for the Control of Autoimmunity. Journal of Biological
Chemistry 288:25490-25499, 2013.

166

Watanabe, Tomonobu M.; Fujii, Fumihiko; Jin, Takashi; Umemoto,
Eiji; Miyasaka, Masayuki; Fujita, Hideaki; Yanagida, Toshio. FourDimensional Spatial Nanometry of Single Particles in Living Cells
Using Polarized Quantum Rods. Biophysical Journal 105:555-564,
2013.

167

Watanabe, Tomonobu M.; Imada, Katsumi; Yoshizawa, Keiko;
Nishiyama, Masayoshi; Kato, Chiaki; Abe, Fumiyoshi; Morikawa,
Takamitsu J.; Kinoshita, Miki; Fujita, Hideaki; Yanagida, Toshio. Glycine
Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic
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168

Wing, James B.; Sakaguchi, Shimon. Foxp3 T-reg cells in humoral
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169

Yamaguchi, Tomoyuki; Kishi, Ayumi; Osaki, Motonao; Morikawa,
Hiromasa; Prieto-Martin, Paz; Wing, Kajsa; Saito, Takashi; Sakaguchi,
Shimon. Construction of self-recognizing regulatory T cells fromconventional T cells by controlling CTLA-4 and IL-2 expression.
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170

Yamanaka, Masahito; Saito, Kenta; Smith, Nicholas I.; Kawata,
Satoshi; Nagai, Takeharu; Fujita, Katsumasa. Saturated excitation offluorescent proteins for subdiffraction-limited imaging of living cells
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171

Yamanaka, Masahito; Yonemaru, Yasuo; Kawano, Shogo; Uegaki,
Kumiko; Smith, Nicholas I.; Kawata, Satoshi; Fujita, Katsumasa.
Saturated excitation microscopy for sub-diffraction-limited imaging
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172

Yamazaki, Chihiro; Sugiyama, Masanaka; Ohta, Tomokazu; Hemmi,
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173

Yano, Tomoki; Matsui, Takeshi; Tamura, Atsushi; Uji, Masami; Tsukita,
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174

Yao, Bo; Zheng, Dandan; Liang, Shide; Zhang, Chi. Conformational
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175

Yasuhara, Noriko; Yamagishi, Ryosuke; Arai, Yoshiyuki; Mehmood,Rashid; Kimoto, Chihiro; Fujita, Toshiharu; Touma, Kenichi; Kaneko,
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176

Yokota, Shumpei; Imagawa, Tomoyuki; Mori, Masaaki; Miyamae,
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177

Yonemaru, Yasuo; Yamanaka, Masahito; Smith, Nicholas I.; Kawata,
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178

Yoshida, Yuji; Tanaka, Toshio. Interleukin 6 and Rheumatoid Arthritis.
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179

Zenker, Stefanie; Panteleev-Ivlev, Julia; Wirtz, Stefan; Kishimoto,Tadamitsu; Waldner, Maximilian J.; Ksionda, Olga; Tybulewicz, Victor
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180

Zhang, Z. -X.; Geng, D. -Y.; Han, Q.; Liang, S. -D.; Guo, H. -R.. The
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181

Zou, Jian; Kawai, Taro; Tsuchida, Tetsuo; Kozaki, Tatsuya; Tanaka,
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115

Research Outputs

Lectures by PIs
Lecturers

116

Meeting

Country

Date

SwedenApr. 4

Toshio Yanagida

7th GREEN Symposium

Japan

Oct. 9USA

Apr. 15Jun Hatazawa

Japan-Russia International Symposium for Medical Exchange

Japan

Oct. 10

Harvard Medical School Committee on Immunology SeminarUSA

Apr. 17

Shimon Sakaguchi

75th Japan Society for Hematology Annual Meeting

Japan

Oct. 11

Annual Meeting of Japanese Society of PediatricJapan

Apr.19Kazuya Kikuchi

9th AFMC International Medicinal Chemistry Symposium (AIMECS 2013)

TaiwanOct. 15

Shimon Sakaguchi

Eberly Distinguished Lectureship in Immunology at Pittsburgh University

USA

Apr. 25

Taroh Kinoshita

Seminar at School of Pharmaceutical Science, Busan National University

Korea

Oct. 16

Cevayir Coban

22nd Turkish Immunology Conference

TurkeyApr. 27

Taroh Kinoshita

Fall International Convention of the Pharmaceutical Society of Korea

Korea

Oct. 17

Ken Ishii

22nd Turkish Immunology Conference

TurkeyApr. 27

Kiyoshi Takeda

FIMSA International Symposium on Autoimmune Diseases

China

Oct. 17

Kiyoshi Takeda

American Association of Immunologists Annual Meeting 2013

USA

May 3

Ken Ishii

7th Annual Vaccine Renaissance Conference

USA

Oct. 18

Shimon Sakaguchi

American Association of Immunologists Annual Meeting 2013

USA

May 3

Toshio Yanagida

Joint Weizmann-MBI Mechanobiology Conference "Dynamic Architecture of Cell and Tissues"

Takashi Saito

American Association of Immunologists Annual Meeting 2013

USA

May 3

Toshio Yanagida

51st Annual Meeting of the Biophysical Society of Japan

Shimon Sakaguchi

Karolinska Research Lectures at Nobel Forum

Shizuo Akira

Emory Vaccine Center Special Seminar

Shizuo Akira
Tadamitsu Kishimoto

Lecturers

Meeting

Country

DateSingapore

Oct. 21Japan

Oct. 27

Cevayir Coban

10th Prof. Dr. Fethi Tezok Microbiology and Infectious Diseases Symposium

TurkeyMay 3

Ken Ishii

7th Vaccine & ISV Congress

Spain

Oct. 28

Kiyoshi Takeda

33rd Korean College of Rheumatology Annual Scientific Meeting

Korea

May 10Atsushi Kumanogoh

EMBO Workshop

FranceOct. 31

Kazuya Kikuchi

Advanced Practical Course in Chemical Biology

Germany

May 16

Toshio Yanagida

Workshop on Modeling Biomolecular Systems in Cellular Environments

Japan

Nov. 1Ken Ishii

21st International Symposium of Macrophage Molecular and Cellular Biology

Japan

May 20

Tadamitsu Kishimoto

100th Anniversary of Kitasato InstituteJapan

Nov. 5

Tsuneyasu Kaisho

Joint International Meeting of the 78th Meeting of the Japanese Society of Interferon and Cytokine
Research and the 21st International Symposium on Molecular CellJapan

May 20Tsuneyasu Kaisho

2013 Agricultural Biotechnology Symposium "Immune Modulation and Bioconvergence"

Korea

Nov. 6

Kiyoshi Takeda

2013 Fall Conference of the Korean Association of Immunologists

Korea

Nov. 7

Tsuneyasu Kaisho

Biology of Macrophage (JSICR-MMCB 2013)

Japan

May 20Tsuneyasu Kaisho

2013 Fall Conference of the Korean Association of Immunologists

Korea

Nov. 7Jun Hatazawa

BRAIN & Brain PET 2013

ChainaMay 20

Shimon Sakaguchi

12th International Xenotransplantation Association Congress 2013

Japan

Nov. 10

Yutaka Hata

IEEE 43rd International Symposium on Multiple-Valued LogicJapan

May 22Ken Ishii2013 Symposium on Complex Biodynamics and Networks

Japan

Nov. 12

Tomohiro Kurosaki

Okinawa Institute of Science and Technology SeminarJapan

May 24

Tomohiro Kurosaki

2013 Oversea Scholar Seminar, Tsinghua University

China

Nov. 14

Toshio Yanagida

8th Symposium of Asian Biophysics Association

Korea

May 26Jun Hatazawa

CJK2013 The 6th CJK Conference on Nuclear Medicine

Korea

Nov. 15

Masaru Ishii

10th Anniversary Symposium of the Hwason Optical Imaging Workshop & Symposium (Plenary)

Korea

May 28Cevayir Coban

International Symposium TCUID 2013

Japan

Nov. 18

Shizuo Akira

78th Cold Spring Harbor Symposium on Quantitative Biology "Immunity & Tolerance"USA

May 30Kazuya Kikuchi

International Symposium TCUID 2013

Japan

Nov. 18

Shimon Sakaguchi

40th IMSUT (The Institute of Medical Science, The University of Tokyo) Founding Commemorative Symposium

Japan

May 31Jun Hatazawa

ICRT2013 8th International Conference on Radiopharmaceutical Therapy

Philippines

Nov. 20

Shimon Sakaguchi

6th International Workshop of Kyoto T Cell ConferenceJapan

Jun. 3Kazuya Kikuchi

2nd Japan-France Coordination Chemistry Symposium

Japan

Nov. 24

Tadamitsu Kishimoto

Charles A. Janeway Jr. MD Memorial Symposium

USA

Jun. 4

Shimon Sakaguchi

41st Japan Society for Clinical Immunology Meeting

Japan

Nov. 27

Tsuneyasu Kaisho

France-Japan Joint Forum “Frontiers in Innate Immunity''

FranceJun. 5

Shimon Sakaguchi

67th Japanese Society of Allergology Meeting

Japan

Nov. 28

Shimon Sakaguchi

Immunology Interest Group Seminar Series in NIH

USA

Jun. 12

Takashi Saito

43rd Annual Scientific Meeting Australasian Society for Immunology

New ZealandDec. 2

Shimon Sakaguchi

Program in Immunology Seminar Series University of Alabama at BirminghamUSAJun. 13

Ken Ishii

Key Lab Medical Molecular Virology Shanghai Medical College, Fudan University

China

Dec. 2

Toshio Yanagida

CREST & PRESTO Joint Symposium for Photonics

Japan

Jun. 20

Tadamitsu Kishimoto

Annual Meeting of Japanese Society of Molecular Biology

Japan

Dec. 3

Shizuo Akira

13th Annual Meeting of the Federation of Clinical Immunology Societies (FOCIS2013)

USA

Jun. 28

Hisashi Arase

Germany-Japan Immunology Seminar 2013

Japan

Dec. 5

Toshio Yanagida

2nd HDP International Symposium on Multi-Level Systems Biology

JapanJun. 28

Tadamitsu Kishimoto

Germany-Japan Immunology Seminar 2013

Japan

Dec. 5

Kazuya Kikuchi

7th International Conference on Materials for Advanced Technologies (ICMAT 2013)

Singapore

Jun. 30

Kiyoshi Takeda

Germany-Japan Immunology Seminar 2013

Japan

Dec. 5

Shimon Sakaguchi

34th Japanese Society of Inflammation and Regeneration Annual Meeting

Japan

Jul. 2

Shimon Sakaguchi

Germany-Japan Immunology Seminar 2013

Japan

Dec. 5

Taroh Kinoshita

3rd Austria/Japan Seminar on Comparative and Developmental Glycobiology

Japan

Jul. 3

Takashi Saito

Germany-Japan Immunology Seminar 2013

Japan

Dec. 5

Taroh Kinoshita

FASEB SRC-Protein Lipidation, Signaling, and Membrane Domains

USA

Jul. 15

Kazuya Kikuchi

International Conference on Emerging Trends in Chemical Science (IETC2013)

India

Dec. 5

Taroh Kinoshita

Seminar at Department of Microbiology, University of Massachusetts

USA

Jul. 18

Shimon Sakaguchi

42th Japanese Society for Immunology Annual Meeting

Japan

Dec. 11

Kazuya Kikuchi

27th Annual Symposium of The Protein Society

USA

Jul. 20

Tsuneyasu Kaisho

42th Japanese Society for Immunology Annual Meeting

Japan

Dec. 11Masahiro Yamamoto

XIVth International Congress of Protistology

CanadaJul. 28

Masaru Ishii

42th Japanese Society for Immunology Annual Meeting

Japan

Dec. 11

Takashi Saito

Frontiers in Immunology ConferenceJapan

Jul. 29Jun Hatazawa

ARCCNM2013 The 12th Annual General Meeting of the ARCCNM

India

Dec. 12

Tomohiro Kurosaki

JSI Immunology Summer School

Japan

Jul. 31

Tomohiro Kurosaki

36th Annual Meeting of the Molecular Biology Society of JapanJapan

Dec. 13

Hisashi Arase

15th JSI Immunology Summer School

Japan

Aug. 1

Tomohiro Kurosaki

Memorial Symposium for Professor Tatsushi Muta

Japan

Dec. 21

Masaru Ishii

FASEB Summer Research ConferenceJapan

Aug. 5

Toshio Yanagida

25th Rheumatism Seminar, NakanoshimaJapan

Dec. 21

Kazuya Kikuchi

15th Asian Chemical Congress (15 ACC)

SingaporeAug.19Masaru Ishii

Gordon Research Conferences

USA

Jan. 12

Tomohiro Kurosaki

5th International Conference on B cell and Autoimmunity

Italy

Aug. 20

Daron M. Standley

Antibody Design, Modeling, and Applications

Japan

Jan. 14

Masaru Ishii

Cold Spring Harbor Conference Asia

China

Aug. 20

Toshio Yanagida

German-Japanese Symposium 2014

Germany

Jan. 16

Tadamitsu Kishimoto

15th International Congress of Immunology

Italy

Aug. 22

Tadamitsu Kishimoto

Keystone Symposia

CanadaJan. 17

Shimon Sakaguchi

15th International Congress of Immunology

Italy

Aug. 22

Shimon Sakaguchi

Keystone Symposia

CanadaJan.17

Shizuo Akira

15th International Congress of Immunology

Italy

Aug. 23

Nicholas Isaac Smith

Annual Meeting, The Physical Society of Republic of China 2014

TaiwanJan. 22

Hisashi Arase

67th Annual Meeting of Tohoku Society of Microbiology

Japan

Aug. 30

Taroh Kinoshita

IUBMB 10th International Symposium on Roles of Eukaryotic Cell Surface Macromolecules

India

Jan. 24

Tomohiro Kurosaki

Immune-related Pathologies: Understanding Leukocyte Signaling and Emerging Therapies – IMPULSE 2013

Hungary

Sep. 2

Tomohiro Kurosaki

World-leading Immunology Workshop at Institute of Development, Aging, and Cancer, Tohoku University

Japan

Jan. 29

Nicholas Isaac Smith

EMSG Ardgour Symposium, Scotland

Scotland

Sep. 3

Tadamitsu Kishimoto

Senri Life Science International Symposium “Innate Immunity, Cytokines, and Immune Regulation”

Japan

Jan. 31

Rikinari Hanayama

86th Annual Meeting of the Japanese Biochemical SocietyJapan

Sep. 12

Shimon Sakaguchi

Senri Life Science International Symposium “Innate Immunity, Cytokines, and Immune Regulation”

Japan

Jan. 31

Shizuo Akira

Annual SAB Meeting Dallas PO1 Genetic Analysis of Resistance to Viral Infection

USA

Sep. 16

Tomohiro Kurosaki

Keystone Symposia: Biolgy of B Cell Reponses

USA

Feb. 11

Ken Ishii

5th International Conference on Crossroads between Innate and Adaptibe Immunity

GreeceSep. 21

Rikinari Hanayama

10th Miyazaki Science CampJapan

Feb. 14Jun Hatazawa

5th Asia-Pacific Symposium on Radiochemistry '13

Japan

Sep. 25

Toshio Yanagida

GCOE-CNR Workshop: Past and Future Directions of Cognitive Neuroscience Robotics

Japan

Feb. 24

Shimon Sakaguchi

4th International GK Symposium Regulators of Adaptive Immunity

Germany

Sep. 27

Nicholas Isaac Smith

Japan Taiwan Bilateral Conference on Biomedical and Plasmonic Imaging

TaiwanFeb. 26

Takashi Saito

4th International GK Symposium Regulators of Adaptive Immunity

Germany

Sep. 27

Shimon Sakaguchi

13th Japanese Society for Regenerative Medicine Meeting

Japan

Mar. 4

Tadamitsu Kishimoto

International Cytokine Meeting

USA

Sep. 29

Shimon Sakaguchi

10th German Society of Immunology Spring School on Immunology

Germany

Mar. 9

Shimon Sakaguchi

4th International Workshop on Humanized Mice

Korea

Sep. 30

Shizuo Akira

8th World Immune Regulation Meeting-VIII2014

Switzerland

Mar. 19

Tadamitsu Kishimoto

Genentech Institute Seminar

USA

Oct. 1Ken Ishii

World Vaccine Congress & Expo

USA

Mar. 24

Masaru Ishii

72nd Annual Meeting of the Japanese Cancer Association

Japan

Oct. 3

Tadamitsu Kishimoto

Conference on Advances in Targeted Therapies

GreeceMar. 26

Hisashi Arase

13th Kyoto Clinical Immunology Seminar

Japan

Oct. 5Jun Hatazawa

1st KURRI International Workshop on Neutron Capture Therapy

Japan

Mar. 27

117

Research Outputs

Access Map
SUITA CAMPUS Osaka University

Graduate School of Medicine
Handai- guchi
Ibaraki Mihogaoka

Senri Gate
Osaka Monorail
Handai-Byoin-Mae Station
Handai Honbumae
Graduate School of Engineering
Hankyu Railway
Kita-Senri Station

Main Gate

Access from the nearest station
By Train
15 min. walk east from Kita-Senri Station on the
Hankyu Senri Line.
By Monorail
25 min. walk from Handai Byoin Mae Station on
the Osaka Monorail.
By Hankyu Bus
Option1:
Get on the bus bound for "Handai Honbu Mae" or"Ibaraki Mihogaoka" from Senri-Chuo Station.Get
off at Handai Honbu Mae, walk 15 min.
Option2:
Get on the bus bound for "Onohara Higashi, Fuji
Kasai" from Senri-Chuo tation. Get off at Handaiguchi, walk 5 min.
By Kintetsu Bus
Get on the bus bound for "Handai Honbu Mae"or "Ibaraki Mihogaoka" from Hankyu Ibarakishi
Station (via JR Ibaraki Station). Get off at Handai
Honbu Mae, walk 15 min.
118

Graduate School of Frontier Biosciences

Kita Fire
Station

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ADDRESS 2Fl. IFReC Research Building, Osaka University
3-1 Yamadaoka, Suita, 565-0871, Osaka, JapanTEL
General Affairs Section: +81-6-6879-4275
Accounting Section: +81-6-6879-4917
Research Planning & Management Office: +81-6-6879-4777
FAX+81-6-6879-4272
E-mailifrec-office@ifrec.osaka-u.ac.jp
119

WPI Immunology Frontier Research Center FY 2013

Edit : Research Planning & Management Office, IFReC
Published in June, 2014

O s a k a U n i ve r s i t y

Copyright : Immunology Frontier Research Center (IFReC ), Osaka University

World Premier International
Research Center

Osaka University

Immunology
Frontier
ResearchCenterAnnual Reportof IFReC
FY 2013