International Immunology, Vol. 21, No. 4, pp. 313–316 ª The Japanese Society for Immunology. 2009.

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IMMUNOLOGY IN JAPAN

The study of innate immunity in Japan: a historical

perspective
Tsuneyasu Kaisho1 and Kiyoshi Takeda2,3
1
Laboratory for Host Defense, RIKEN Research Center for Allergy and Immunology, 1-7-22, Suehiro-cho, Tsurumi-ku, Yokohama,
Kanagawa 230-0045, Japan
2
Laboratory of Immune Regulation, Department of Microbiology and Immunology, Graduate School of Medicine, Osaka
University and 3World Premier International Immunology Frontier Research Center, 2-2 Yamada-oka, Suita, Osaka

REVIEW
565-0871, Japan

Keywords: antigen-presenting cell, CpG DNA, lipopolysaccharide, phagocytes, Toll-like receptor

Abstract
Innate immunity is key for host defense in a vast range of organisms, from invertebrates to
vertebrates. A number of Japanese scientists have made significant contributions to the clarification
of innate immune mechanisms and have applied this knowledge to treatments for various diseases
including cancer, allergy and autoimmune disorders. Here, we present an overview of the

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development of innate immunology by highlighting several Japanese contributors.

Introduction
The immune response is traditionally and conveniently cate-
gorized into two systems—innate immunity and adaptive
immunity—although these systems interact and overlap exten-
sively. The concept of innate immunity has its origin at the end
of 19th century, following experiments on the larvae of starfish
by a Russian microbiologist and zoologist, Mechnikov (1).
Adaptive immunity is based on recognition of antigen by di-
verse repertories of receptors. Antibodies are produced by
B cells and their diversity is established by the somatic gene
rearrangement system (2), which also works in generating
the repertoire of TCRs, and enables two important features of
the immune system in advanced vertebrates—adaptation and
memory. Innate immunity plays critical roles in host defense in
all plant and animal species and cooperates with adaptive im-
munity in the more-advanced organisms. To a large extent, the
development of immunology has been driven by a desire to
clarify the functions of adaptive immune cells rather than in-
nate immune cells; however, it has become increasingly clear
that innate immunity is more sophisticated than was once
thought and crucially influences adaptive immunity.
It is obvious that immunologists all over the world have
illuminated the mechanisms involved in innate immunity, but
here we focus on the contribution of several Japanese
immunologists to the topic.
Phagocytes: from Mechnikov’s time to the 21st century
The cells that Mechnikov identified in starfish can incorpo-
rate and digest invading micro-organisms. Cells that do this
are called phagocytes and the engulfment phenomenon is
called phagocytosis; this is just one of a wide range of
mechanisms used in innate immunity.
Innate immune cells such as phagocytic macrophages
can migrate toward lesions and are critically involved in
establishing inflammation. In the 1960s, Hideo Hayashi at
Kumamoto University characterized several factors that can
attract various kinds of leucocytes. For example, leukoegre-
sin was identified as a chemotactic factor for polymorphonu-
clear leucocytes (3). These soluble factors are nowadays
known as chemokines (4).
Phagocytes such as macrophages engulf not only micro-
organisms (non-self) but also self-derived apoptotic cells. In
the 1990s and the early years of this century, Shigekazu
Nagata (5), while working at Osaka Bioscience Institute and
Osaka University, questioned how apoptotic cells are recog-
nized by phagocytes and revealed the mechanism. He iden-
tified several molecules of phagocytes that recognize an
‘eat-me’ signal, which is expressed on the surface of dying
cells. He further clarified how engulfed apoptotic cells, and
especially self-derived DNA, are processed within phago-
cytes. The disturbance of this process can result in autoim-
mune disorders such as arthritis.
An important function of innate immunity in vertebrates is
antigen presentation to T cells. This antigen-presenting ability
is critical for linking innate and adaptive immunity. Antigen
presentation depends on adherent non-lymphoid lineage cells
and phagocytic activity. Therefore, macrophages had been

Correspondenc to: T. Kaisho; E-mail: tkaisho@rcai.riken.jp, K. Takeda; Received 26 January 2009, accepted 30 January 2009
E-mail: ktakeda@ongene.med.osaka-u.ac.jp
Advance Access publication 26 February 2009
314 Innate immunity in Japan
considered to be major players as antigen-presenting cells
(APCs). The pioneering work of Steinman and Cohn (6) at
the Rockefeller University in New York showed APC activity in
non-adherent and non-phagocytic cells, which are now called
dendritic cells (DCs). Kayo Inaba and Shigeru Muramatsu (7)
at Kyoto University identified non-macrophage cells as critical
cell components for in vitro antibody response. Then Kayo
Inaba, through fruitful collaboration with Steinman’s group,
contributed to clarifying DC functions. She revealed that DCs
can be generated by an in vitro culture from bone marrow
precursors.
Using innate immunity to combat cancer
Around the same time that Mechnikov described phago-
cytes, an American surgeon called Coley developed immu-
notherapy against cancer (8). He observed that a cancer
patient who suffered from severe infection with Streptococcus
pyogenes recovered from the cancer. On the basis of
this experience, Coley succeeded in shrinking tumors by
injecting a brew of S. pyogenes directly into tumors. This
success led to the generation of the Coley vaccine that is
composed of dead S. pyogenes and dead Serratia marces-
cens bacteria. It is now well recognized that production of
tumor necrosis factor during the infection contributes to the
curative effects on cancers.
Yuichi Yamamura is another pioneer in developing a thera-
peutic maneuver, based on innate immunity, for cancers
(Fig. 1). He was mainly involved in clarifying the mecha-
nisms by which Mycobacterium tuberculosis or related sub-
stances cause disease and in applying knowledge about
mycobacteria to help treat cancers. In the 1950s, the pulmo-
Fig. 1. Yuichi Yamamura.
nary cavity in tuberculosis patients was considered to be
generated by live mycobacteria, like a house being eaten
away by termites. While working as a clinician at the
Toneyama Hospital in Osaka, Yamamura demonstrated
that dead mycobacteria or lipoprotein extracts from myco-
bacteria can induce cavities similar to those in patients with
tuberculosis and that this cavity formation depends on
T-cell-mediated immunity (9).
These findings can be now interpreted as follows: micro-
organism-derived molecular components, which include
some Toll-like receptor (TLR) ligands (see below), can in-
duce the cavity formation by stimulating innate immune cells
to activate T cell-mediated adaptive immune responses. At
Kyushu and Osaka Universities, Yamamura has further ex-
tended his studies by finding a potent macrophage-activat-
ing ability in the cell wall extracts of a mycobacterium,
Nocardia rubra, and applying them for certain cancers. He
also made a great contribution to establishing the Japanese
Society for Immunology, which started in 1970, and organiz-
ing Fifth International Congress of Immunology in Kyoto in
1983.
Tohru Tokunaga et al. (10) at the National Institute of Infec-
tious Diseases in Tokyo first found that immunostimulatory
activity of the extracts from a Mycobacterium bovis strain,
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bacillus Calmette–Guérin (BCG), can be ascribed to the
bacterial genomic DNA (Fig. 2). DNA purified from BCG
inhibited tumor growth, augmented NK cell activity and in-
duced type I IFNs from murine spleen cells and human pe-
ripheral blood lymphocytes. Then Tohru Tokunaga et al.
clarified that the activity required the palindromic sequences
including the 5#-CG-3’ motif (11). They further found that
the activity can be detected widely in DNAs from micro-
organisms and invertebrates, but not in DNAs from verte-
brates or plants and suggested that this differential activity
Fig. 2. Tohru Tokunaga.
 Innate immunity in Japan 315
depends on the frequency of 5#-CG-3’ motifs (12). This work
was performed before Krieg’s group reported that bacterial
DNA containing the CpG motif can directly activate B cells
(13) Thus, Tokunaga has illuminated the concept of immu-
nostimulatory DNA, which now shows a great promise as an
anti-allergic or anti-cancer vaccine.

Evolving innate immunity: what Toll has told us

Invertebrates do not have adaptive immunity, but this does
not mean that host defense systems are undeveloped in
invertebrates. Invertebrates have instead acquired an ad-
vanced type of host defense by evolving innate immunity,
and Japanese investigators were heavily involved in delin-
eating the innate immune responses in various invertebrates.
The horseshoe crab is a marine arthropod found most
commonly in East Asia and North America. This arthropod,
which has been called a living fossil, is exceptionally useful
for biochemical analysis on proteins in the hemolymph be- Fig. 3. Shizuo Akira.
cause several hundreds of milliliters of hemolymph can be
taken from one horseshoe crab without threatening its life.
The hemolymph contains a single species of hemocytes that
until the 1990s. In 1998, the responsible mutation for LPS re-
is extremely sensitive to bacterial endotoxin, LPS. In the
fractoriness in mice was found to be located in the gene
1990s, Sadaaki Iwanaga at Kyushu University clarified the

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locus encoding Tlr4, which is structurally related to Drosophila
molecular mechanisms of hemolymph coagulation cascades
Toll (18). This ignited the interest of lots of immunologists
in the horseshoe crab (14). These cascades sense micro-
and microbiologists. Mammalian TLRs, which are now known
organism-derived components including LPS and (1/3)-b-
to consist of ;10 members, have their specific roles in sens-
D-glucan and are essential for immobilization of microbial
ing infection by micro-organisms through the detection of
pathogens, which are subsequently killed by anti-microbial
microbial components such as lipids or nucleic acids. Since
peptides.
the end of the 20th century, Shizuo Akira, while working at
The limulus test is a sensitive LPS detection system widely
Hyogo College of Medicine and Osaka University, has clari-
used in experiments and clinical medicine and is based on
fied most of their functions and signaling mechanisms
the work of Iwanaga. It is to be noted that the components
mainly through generation and characterization of knockout
of the cascades exhibit homology to proteins that act
mice (Fig. 3, reviewed in this issue). These studies have also
upstream of Toll. Toll is a Drosophila transmembrane recep-
elucidated the molecular mechanisms behind adjuvants’
tor that plays critical roles not only in insect development
ability to activate immunity. For example, Akira’s great
but also in anti-microbial immune responses (15). Invading
achievements include identification of the receptor for the
micro-organisms are recognized by soluble factors in the
immunostimulatory DNA which carries the unmethylated
hemolymph and protease cascades are subsequently acti-
CpG motif (19).
vated. Then a host protein, Spaetzle, is cleaved and acti-
vates Toll-mediated signaling that leads to production of
anti-microbial peptides. Iwanaga’s studies provided a frame- Conclusions
work for many cascade reactions in invertebrate innate im-
Thus, a number of Japanese immunologists have contrib-
munity, which contributed later on the investigation of the
uted to the understanding of innate immunity. It is difficult
Drosophila Toll system. Shunji Natori (16) at Tokyo University
and beyond our capacity to refer to all contributions and the
has isolated several other proteins that act in insect immu-
achievements listed here are only the tip of the iceberg. We
nity, including anti-microbial peptides from the flesh fly, Sar-
know that we have much more to learn but we hope that
cophaga peregrina, and, before the discovery of Toll acting
Japanese scientists will continue to play an important role,
on innate immunity and development, demonstrated that
together with our colleagues worldwide, in making these
some insect immunity proteins have dual functions in de-
discoveries.
fense and development.
In the 1990s, Janeway (17) at Yale University stated that ef-
fective immune responses require certain APC-activating sig- Acknowledgements
nals in addition to protein antigens. Those substances are We would like to thank Tadamitsu Kishimoto, Tohru Tokunaga, Kayo
immune adjuvants and he called them the immunologist’s Inaba, Kiyoshi Takatsu, Shin-Ichi Hayashi, Hitoshi Kikutani, Ken
‘dirty little secret’ in that article. Immune adjuvants are required J. Ishii, Tatsushi Muta, and Shoichiro Kurata for helpful advice or
for effective antigen uptake as well as for activating APCs. providing useful information. The photos of Yuichi Yamamura, Tohru
Tokunaga and Shizuo Akira were kindly provided by Tadamitsu
LPS is a representative of the immune adjuvants and has Kishimoto, Tohru Tokunaga and Shizuo Akira, respectively. We feel
been extensively investigated for decades; however, the crit- very sorry not to be able to refer all contributors who have been
ical signal-transducing receptors for LPS were not identified critical to our current understanding of innate immunity.
316 Innate immunity in Japan
Abbreviations
APC antigen-presenting cell
BCG bacillus Calmette–Guérin
DC dendritic cell
TLR Toll-like receptor
TNF tumor necrosis factor
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