Department of
Cardiovascular Medicine
Outline and Research Objectives
In 1998 the Department of Internal Medicine at University of Tokyo hospital was reorganized to the more functional units based on clinical specialties of diseased organs.
Cardiologists from 5 departments of Internal Medicine were unified to the Department of
Cardiovascular Medicine. We at quite an early phase introduced techniques of molecularbiology into the research field of cardiovascular medicine. As a result, we could report following important findings: 1) phenotypic modulation of cardiac or vascular specific proteins
occurs during cardiac hypertrophy or arteriosclerosis. 2) intracellular mechanisms on how
mechanical stress causes cardiac hypertrophy; 3) identification of novel genes expressed
during heart development; 4) development of animal models for cardiovascular diseases byusing genetic engineering techniques (i.e. knockout mice of endothelin-1 and KLF5 genes
provided new findings for the research of this field); 5) therapeutic angiogenesis for experimental myocardial infarction by growth factors was originally introduced in this department; 6) We are developing an electronic patient record system by which prognosis as wellas risk factors of cardiovascular disease can be statistically analyzed. From a research standpoint, our interests range throughout all fields of cardiovascular medicine ranging frommolecular biology to clinical research including genomics. Importantly, our research interests are aimed at making possible new diagnostics and treatment of cardiovascular diseases.
Areas of interest are as follows:
1) Transcriptional regulation in cardiovascular pathogenesis
2) Risk factor analysis of cardiovascular disease3) Pathogenic mechanisms of cardiac hypertrophy and heart failure4) Immunological basis of myocarditis and dilated cardiomyopathy5) MRI in cardiovascular diseases
6) Early diagnosis of ischemic heart disease using radionuclide testing7) Aerobic threshold and cardiac rehabilitation
8) Anti-arrhythmia therapy using atrial remodeling9) Mechanism of post-PTCA restenosis10) Molecular mechanisms of reperfusion injury11) Genetic polymorphisms in cardiovascular disease12) Differentiation of smooth muscle cells13) Cardiac development
14) Gene expression and regulation in cardiomyocytes15) Mouse genetic models of cardiovascular diseases and vascular development
16) Nitric oxide and endothelial function17) Gene therapy of heart failure using cardiac contractile proteins18) Clinical application of vasoactive substances

Faculties and Students
Professor and Chair
Lecturer

Ryozo Nagai, MD, PhD (1999~)
Yasunobu Hirata, MD, PhD

Minoru Ohno, MD, PhD
Yoshinori Seko, MD, PhD
Associate ...............................14
Postdoctoral Fellow ..............11
Graduate student ..................28
Research student.....................7
Secretary .................................4

Past Research and Major Accomplishments
1) Transcriptional regulation of expression ofgenes related to cardiovascular diseases
We have been focusing our research on the role oftranscriptional regulation of gene expression in the
cardiovascular system with a particular interest on
regulation of phenotypic modulation of smooth muscle and cardiac cells. In 1980s and early 1990s, Nagai
et al isolated and characterized three types of smooth157

muscle myosin heavy chains and found that they are
differentially expressed during vascular development.
Nagai et al furthermore isolated a DNA-binding factor,
Krüppel-like factor 5 (KLF5), as a transcription factorof the embryonic isoform of smooth muscle myosin
heavy chain gene (SMemb), whose expression is
induced in phenotypically modulated smooth muscle
cell and cardiac fibroblast. Recently, by developing
knockout mice of KLF5 gene, we have found that
KLF5 is an essential regulator of cardiovascularremodeling which occurs in response to various external stresses (Figure 1). We have further found that differential chemical modifications and protein-protein
interactions regulate this family of factors. We are currently investigating whether low molecular weightcompounds that inhibit functions of KLF5 are clinically applicable as a new therapeutic drug against cardiovascular diseases.
2) Differentiation of smooth muscle cells in vascular lesions
On the contrary to general assumption that neointima cells are derived from medial smooth muscle
cells, we found that bone marrow cells give rise to themajority of smooth muscle cells that contribute toarterial remodeling in models of post-angioplasty
restenosis, graft vasculopathy and hyperlipidemiainduced atherosclerosis. Notably, we found purified
hematopoietic stem cells differentiated into smoothmuscle cells in vitro and in vivo. We proposed thatamong blood cells there may be progenitors ofsmooth muscle cells, which attach to the injured
endothelia, differentiate into smooth muscle cells and158

proliferate, contributing thereby to neointima formation. Our results will also suggest a novel strategy toprevent vascular diseases, targeting mobilization (A),circulation (B), homing (C), differentiation (D) and proliferation (E) of putative smooth muscle progenitor
cells (Figure 2).
3) Establishment of clinical data management system and its practical application to genetic epidemiology in cardiovascular medicine
We have established a clinical data analysis systemon network, which enrolls more than 2000 patients
who underwent coronary angiography in our department. By using this system we can analyze the relationship between the genetic polymorphisms and theclinical of cardiovascular diseases such as coronary
artery diseases (CAD) and ischemic stroke. We
demonstrated that the alanine/valine (A/V) polymorphism in the gene of 5,10-methylenetetrahydrofolate
reductase (MTHFR) geneis a significant genetic risk
factor for CAD and ischemic stroke. This genetic variant is a more powerful predictor of atherosclerotic diseases, when the plasma folate levels are relatively low.
4) Development of mouse genetic models of cardiovascular diseases
Our research interests include the pathophysiological assessment of cardiovascular diseases by micegene engineering approach. We have generated
knockout and transgenic mice of adrenomedullin,
ADAMTS-1, endothelin-1 and KLF5. We analyzed therole of these genes and found that their expression isessential for normal growth of mice.

Figure 3 summarizes our strategy for clinical application of basic research on cardiovascular disease.

Current Research
1) Transcription factor such as Krüppel-like factors in pathogenic mechanisms.
Modulation of transcription factors may lead to a
new therapeutic strategy for cardiovascular disease.
Ongoing studies include using proteomics approaches
to understand the diverse protein-protein interactions
and regulation of these Krüppel-like factors. We havealready identified a novel repressor protein among
others. Another focus of research surrounds understanding the transcriptional regulation of these
Krüppel-like factors in the context of chromatin
which is necessary to understand how transcription
occurs in humans.
2) Bone marrow-derived vascular progenitor cells(1) Identification of bone marrow fraction contributing to atherosclerosis
To determine which fraction of bone marrow cells
contribute to the pathogenesis of vascular diseases,
we are reconstituting bone marrow of the lethally
irradiated recipient mice with the hematopoietic stem
cells from LacZ mouse and the mesenchymal stem
cells from GFP mouse. We are applying following vascular injury models to bone marrow-reconstitution
mice: mouse femoral injury model, heterotopic cardiac
transplantation model and atherosclerotic model.
(2) Characterization of bone marrow-derived
vascular progenitor cells
By using cell sorter, we are identifying which frac-tion of peripheral blood cells effectively differentiates
into smooth muscle cells. We are determining the cellsurface marker to isolate the putative bone marrowderived smooth muscle progenitor cells. Using bonemarrow transplantation mice, we are identifying theeffects of hyperlipidemia and drugs on mobilization,
homing, differentiation of the progenitor cells in various models of vascular injury described above3) Establishment of clinical data management system.
To overcome the difficulty of filing complicatedclinical parameters for genetic association studies, weare accumulating clinical data and constructing ouroriginal database system. For genetic analyses, written informed consent to participate was obtained
from all the in-patients in our department. DNA samples obtained from the participants have reachedapproximately one thousand. Using them, we analyzed over 50 genetic polymorphisms implicated to be
associated with atherosclerotic diseases. Among plenty of SNPs analyzed in our study, we showed thepolymorphisms in the MMP-1 and MMP-3 promoters
are associated with disease susceptibility to myocardial infarction. Also, our database system supports
several ongoing clinical epidemiological studies suchas J-CAD.
4) Genetic engineering for analysis of the role ofendogenous protein
We are analyzing the in vivo function of KLF5
using knockout mice. In response to external stress,

159

KLF5 knockout mice exhibited diminished levels ofarterial wall thickening, cardiac hypertrophy, interstitial fibrosis and that angiotensin II induced expression of KLF5, which in turn activated growth factor
expression. KLF5 thus appears to be a key element
linking external stress and cardiovascular remodeling.

Future Prospects
1) Dissection of the pathogenic mechanisms whereby
the Krüppel-like factors are involved in cardiovascular disease will allow us to understand thediverse mechanisms by which this family of factors are involved in transcriptional regulation. Bysuch, we envision that targeted drug design tomodulate these factors in this context will enable
us to propose a new therapeutic approach to cardiovascular disease (e.g. pin-point drug design).
2) Using our database system we have followed all
the cases periodically, thus we can get prospective
clinical data such as major cardiovascular eventsand responsiveness to treatment such as drugs andpercutaneous coronary intervention. In future, wewill be able to obtain several evidences from ourdatabase system and genetic analyses, resulting inclinical application of such evidences in prediction
or prevention of major cardiovascular diseases.
3) We have shown that KLF5 is a crucial determinantof the cellular response to cardiovascular injury,
playing a key role in mediating cardiovascularremodeling. We are now analyzing the effect ofcompounds, which could modulate KLF5 function
to control remodeling in atherosclerosis and heart
failure.
4) Our studies will show that bone marrow cells
including hematopoietic stem cells have the potential to give rise to vascular progenitor cells thathome in the damaged vessels, differentiate into
smooth muscle cells, and proliferate thereby contributing to vascular remodeling. We expect thatatherogenic factors such as hyperlipidemia, inflammation, cytokines, and smoking will facilitate thekinetics of smooth muscle progenitors. Our findings will provide the basis for the development of
new diagnostic strategies to predict atherosclerosis
by quantifying the circulating progenitors.

Research Grants
Ryozo Nagai (1997-2001)
Grant from the Organization for Pharmaceutical
Safety and ResearchRole of Klotho gene and its clinical application in cardiovascular disease

160

Ryozo Nagai (1999-2001)
Grant from the Ministry of Health, Labour andWelfareResearch on elucidation of the molecular mechanisms of specific diseases
Ryozo Nagai (2000-2002)
Grant from the Ministry of Education, Culture,
Sports, Science and Technology
Development of a new prognostic evaluation systemand treatment for patients with cardiovascular disease based on gene polymorphisms
Ryozo Nagai (2000-2002)
Grant from the Ministry of Health, Labor andWelfareResearch on development of angiogenesis and vascular protection treatment
Ryozo Nagai (2002-2006)
Grant from the Ministry of Education, Culture,
Sports, Science and Technology
Molecular mechanism and organ remodeling: genetranscription and cell-cell interaction in mesenchymal cells

Select Publications
Diversity of smooth muscle myosin heavy chain
isoforms
1. Nagai R, Larson DM, Periasamy M. Characterizationof a mammalian smooth muscle myosin heavy chain
cDNA and its expression in various smooth muscle
types. Proc Natl Acad Sci USA 85: 1047-1051, 1988.
2. Nagai R, Kuro-o M, Babij P, Periasamy M.
Identification of two types of smooth muscle myosin
heavy chain isoforms by cDNA cloning andimmunoblot analysis. J Biol Chem 264: 9734-9737,
1989.
3. Kuro-o M, Nagai R, Tsuchimochi H, Katoh H, Yazaki
Y, Ohkubo A, Takaku F. Developmentally regulated
expression of vascular smooth muscle myosin heavy
chain isoforms. J Biol Chem 264: 18272-18275, 1989.
4. Kuro-o M, Nagai R, Tsuchimochi H, Katoh H, Tsai RC, Yazaki Y, Ohkubo A, Takaku F. cDNA cloning of amyosin heavy chain isoform in embryonic smoothmuscles and its expression during vascular development and in arteriosclerosis. J Biol Chem 266: 37683773, 1991.
5. Kim H-S, Aikawa M, Kimura K, Kuro-o M, Nakahara
K, Suzuki T, Katoh H, Okamoto E, Yazaki Y, Nagai R.
Ductus arteriosus: advanced differentiation ofsmooth muscle cells demonstrated by myosin heavy
chain isoform expression in rabbits. Circulation 88:
1804-1810, 1993.
6. Aikawa M, Nalla Sivam P, Kuro-o M, Kimura K,
Nakahara K, Takewaki S, Yamaguchi H, Yazaki Y,
Periasamy M, Nagai R. Human smooth muscle
myosin heavy chain isoforms as molecular markers
for vascular development and atherosclerosis. Circ
Res 73: 1000-1012, 1993.

7. Suzuki T, H-S Kim, Kurabayashi M, Hamada H, Fujii
H, Aikawa M, Watanabe M, Yazaki Y, Nagai R
Preferential differentiation of P19 mouse embryonalcarcinoma cells into smooth muscle cells: use ofretinoic acid and antisense against the central nervous system-specific POU transcription factor Brn-2.
Circ Res 78:395-404,1996.

tribute to atherosclerosis. Nature Med. 7: 382-383,
2001.
9. Sata M, Saiura A, Kunisato A, Tojo A, Okada S,
Tokuhisa T, Hirai H, Makuuchi M, Hirata Y, Nagai R.
Hematopoietic stem cells differentiate into vascular
cells that participate in the pathogenesis of atherosclerosis. Nature Med. 8: 403-409, 2002.

Regulation of smooth muscle cell differentiationand cardiovascular remodeling
1. Watanabe M, Sakomura Y, Kurabayashi M, Manabe
I, Aikawa M, Kuro-o M, Suzuki T, Yazaki Y, Nagai R
Structure and characterization of the 5’-Flanking
Region of the mouse smooth muscle myosin heavy
chain (SM1/2) gene. Circ Res 78: 978-989, 1996.
2. Manabe I, Kurabayashi M, Shimomura Y, Kuro-o M,
Watanabe N, Watanabe M, Aikawa M, Suzuki T,
Yazaki Y, Nagai R Isolation of the embryonic form ofsmooth muscle myosin heavy chain (SMemb/NMHCB) gene and characterization of its 5’-flanking region.
Biochem Biophys Res Commun 239:598-605, 1997.
3. Watanabe N, Kurabayashi M, Shimomura Y, KawaiKowase K, Hoshino Y, Manabe I, Watanabe M,
Aikawa M, Kuro-o M, Suzuki T, Yazaki Y, Nagai R
BTEB2, a Krppel-like transcription factor, regulates
expression of the SMemb/Nonmuscle myosin heavy
chain B (SMemb/NMHC-B) gene Circ Res 85: 182191, 1999.
4. Kawai-Kowase K, Kurabayashi M, Hoshino Y,
Ohyama Y, Nagai R. Transcriptional activation of thezinc finger transcription factor BTEB2 gene by Egr-1
through mitogen-activated protein kinase pathways
in vascular smooth muscle cells Circ Res 85: 787-795,
1999.
5. Hoshino Y, Kurabayashi M, Kanda T, Hasegawa A,
Sakamoto H, Okamoto Ei, Kowase K, Watanabe N,
Manabe I, Suzuki T, Nakano A, Takase Si, Wilcox
JN, Nagai R. Regulated expression of the BTEB2 transcription factor in vascular smooth muscle cells :analysis of developmental and pathological expression profiles shows implications as a predictive factor for restenosis. Circulation 102: 2528-2534, 2000.
6. Sekiguchi K, Kurabayashi M, Oyama Y, Aihara Y,
Tanaka T, Sakamoto H, Hoshino Y, Kanda T,
Yokoyama T, Shimomura Y, Iijima H, Ohyama Y,
Nagai R. Homeobox protein hex induces SMemb/
Nonmuscle myosin heavy chain-B gene expression
through the cAMP-responsive element. Circ Res 88:
52-28, 2001.
7. Shindo T, Manabe I, Fukushima Y, Tobe K, Aizawa
K, Miyamoto S, Kawai-Kowase K, Moriyama N, Imai
Y, Kawakami H, Nishimatsu H, Ishikawa T, Suzuki
T, Morita H, Maemura K, Sata M, Hirata Y, Komukai
M, Kagechika H, Kadowaki T, Kurabayashi M, Nagai
R. Krppel-like zinc-finger transcription factor
KLF5/BTEB2 is a target for angiotensin II signaling
and an essential regulator of cardiovascular remodeling. Nature Med. 8:856-873, 2002.
8. Saiura A, Sata M, Hirata Y, Nagai R, Makuuchi M.
Circulating smooth muscle progenitor cells con-Identification and function of anti-aging geneklotho1. Kuro-o M, Matsumura Y, Aizawa H, Kawaguchi H,
Suga T, Utsugi T, Ohyama Y, Kurabayashi M,
Kaname T, Kume E, Iwasaki H, Iida A, Shiraki-Iida T,
Nishikawa S, Nagai R, Nabeshima Y. Mutation of themouse Klotho gene leads to a syndrome resembling
ageing. Nature 390: 45-51, 1997.
2. Saito Y, Yamagishi T, Nakamura T, Ohyama Y,
Aizawa H, Suga T, Matsumura Y, Masuda H,
Kurabayashi M, Kuro-o M, Nabeshima Y, Nagai R.
Klotho protein protects against endothelial dysfunction. Biochem Biophys Res Commun 248: 324-329,
1998.
3. Aizawa H, Saito Y, Nakamura T, Inoue M, Imanari T,
Ohyama Y, Matsumura Y, Masuda H, Oba S, Mise N,
Kimura K, Hasegawa A, Kurabayashi M, Kuro-o M,
Nabeshima Y, Nagai R. Downregulation of the klotho
gene in the kidney under sustained circulatory stressin rats. Biochem Biophys Res Commun 249: 865-871,
1998.
4. Suga T, Kurabayashi M, Sando Y, Ohyama Y, Maeno
T, Maeno Y, Aizawa H, Matsumura Y, Kuwaki T,
Kuro-o M, Nabeshima Y, Nagai R. Disruption of theklotho gene causes pulmonary emphysema in mice:
defect in maintenance of pulmonary integrity during
postnatal life Am J Respir Cell Mol Biol 22: 26-33,
2000.
5. Nagai R, Saito Y, Ohyama Y, Aizawa H, Suga T,
Nakamura T, Kurabayashi M, Kuro-o M Endothelial
dysfunction in the klotho mouse and downregulation of klotho gene expression in various animalmodels of vascular and metabolic diseases. Cell Mol
Life Sci 57: 738-746, 2000.
6. Saito Y, Nakamura T, Ohyama Y, Suzuki T, Iida A,
Shiraki-Iida T, Kuro-o M, Nabeshima Y, Kurabayashi
M, Nagai R. In vivo klotho gene delivery protects
against endothelial dysfunction in multiple risk factor syndrome. Biochem Biophys Res Commun
276:767-772, 2000.
7. Mitani H, Ishizaka N, Aizawa T, Ohno M, Usui S,
Suzuki T, Amaki T, Mori I, Nakamura Y, Sato M,
Nangaku M, Hirata Y, Nagai R. In vivo klotho genetransfer ameliorates angiotensin II-induced renal
damage. Hypertension. 39:838-843, 2002.
Intracellular signaling and transcription factors ofcardiac hypertrophy
1. Kojima M, Shiojima I, Yamazaki T, Komuro I, Nagai
R, Yazaki Y. Angiotensin II receptor antagonist
TCV116 regresses hypertensive left ventricular
hypertrophy in vivo and inhibits intracellular signal-161

ing pathway of stretch-mediated cardiomyocyte
hypertrophy in vitro. Circulation 89: 2204-2211,
1994
2. Shimoyama M, Hayashi D, Takimoto E, Zou Y, Oka
T, Uozumi H, Kudoh S, Shibasaki F, Yazaki Y, Nagai
R, Komuro I. Calcineurin plays a critical role in pressure overload-induced cardiac hypertrophy.
Circulation. 100:2449-2454, 1999.
3. Zhu W, Zou Y, Aikawa R, Harada K, Kudoh S,
Uozumi H, Hayashi D, Gu Y, Yamazaki T, Nagai R,
Yazaki Y, Komuro I. MAPK superfamily plays animportant role in daunomycin-induced apoptosis ofcardiac myocytes. Circulation. 100:2100-2107,1999.
4. Aikawa R, Nawano M, Gu Y, Katagiri H, Asano T,
Zhu W, Nagai R, Komuro I. Insulin prevents cardiomyocytes from oxidative stress-induced apoptosis
through activation of PI3 kinase/Akt. Circulation.
102: 2873-2879, 2000.
5. Hiroi Y, Kudoh S, Monzen K, Ikeda Y, Yazaki Y,
Nagai R, Komuro I. Tbx5 associates with Nkx2-5 andsynergistically promotes cardiomyocyte differentiation. Nature Genet. 28:276-280, 2001.
6. Hosoda T, Monzen K, Hiroi Y, Oka T, Takimoto E,
Yazaki Y, Nagai R, Komuro I. A novel myocyte-specific gene Midori promotes the differentiation of
P19CL6 cells into cardiomyocytes. J Biol Chem.
276:35978-35989, 2001.
7. Uozumi H, Hiroi Y, Zou Y, Takimoto E, Toko H, Niu
P, Shimoyama M, Yazaki Y, Nagai R, Komuro I.
gp130 plays a critical role in pressure overloadinduced cardiac hypertrophy. J Biol Chem. 276:
23115-23119, 2001.
8. Zou Y, Hiroi Y, Uozumi H, Takimoto E, Toko H, Zhu
W, Kudoh S, Mizukami M, Shimoyama M, Shibasaki
F, Nagai R, Yazaki Y, Komuro I. Calcineurin plays acritical role in the development of pressure overloadinduced cardiac hypertrophy. Circulation. 104:97101, 2001.
9. Zou Y, Hiroi Y, Uozumi H, Takimoto E, Toko H, Zhu
W, Kudoh S, Mizukami M, Shimoyama M, Shibasaki
F, Nagai R, Yazaki Y, Komuro I. Calcineurin plays acritical role in the development of pressure overloadinduced cardiac hypertrophy. Circulation. 104:97101, 2001.
10. Takimoto E, Yao A, Toko H, Takano H, Shimoyama
M, Sonoda M, Wakimoto K, Takahashi T, Akazawa
H, Mizukami M, Nagai T, Nagai R, Komuro I. Sodium
calcium exchanger plays a key role in alteration ofcardiac function in response to pressure overload.
FASEB J 16:373-378, 2002.

Cardiovascular pathophysiology
1. Kurihara Y, Kurihara H, Suzuki H, Kodama T,
Maemura K, Nagai R, Oda H, Kuwaki T, Cao WH,
Kamada N, Jishage K, Ouchi Y, Azume S, Toyoda Y,
Ishikawa T, Kumada M, and Yazaki Y. Elevatedblood pressure and craniofacial abnormalities in mice
deficient in endothelin-1. Nature 368: 703-710, 1994.
2. Kuro-o M, Hanaoka K, Hiroi Y, Noguchi T, Fujimori
Y, Takewaki S, Hayasaka M, Katoh H, Miyagishi A,

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4.

5.

6.

7.

8.

Nagai R, Yazaki Y, Nabeshima Y. Salt-sensitinve
hypertension in transgenic mice overexpressing sodium-proton exchanger. Circ Res 76: 148-153, 1995.
Shindo T, Kurihara H, Kuno K, Yokoyama H, Wada
T, Kurihara Y, Imai T, Wang Y, Ogata M, Nishimatsu
H, Moriyama N, Oh-hashi Y, Morita H, Ishikawa T,
Nagai R, Yazaki Y, Matsushima K. ADAMTS-1: ametalloproteinase-disintegrin essential for normal
growth, fertility, and organ morphology and function. J Clin Invest. 105:1345-1352, 2000.
Shindo T, Kurihara H, Maemura K, Kurihara Y,
Kuwaki T, Izumida T, Minamino N, Ju KH, Morita H,
Oh-hashi Y, Kumada M, Kangawa K, Nagai R, Yazaki
Y. Hypotension and resistance to lipopolysaccharideinduced shock in transgenic mice overexpressing
adrenomedullin in their vasculature. Circulation.
101:2309-2316, 2000.
Shindo T, Kurihara Y, Nishimatsu H, Moriyama N,
Kakoki M, Wang Y, Imai Y, Ebihara A, Kuwaki T, Ju
KH, Minamino N, Kangawa K, Ishikawa T, Fukuda
M, Akimoto Y, Kawakami H, Imai T, Morita H,
Yazaki Y, Nagai R, Hirata Y, Kurihara H. Vascularabnormalities and elevated blood pressure in micelacking adrenomedullin gene. Circulation. 104:19641971, 2001.
Morita H, Kurihara H, Yoshida S, Saito Y, Shindo T,
Oh-Hashi Y, Kurihara Y, Yazaki Y, Nagai R. Dietinduced hyperhomocysteinemia exacerbates neointima formation in rat carotid arteries after balloon
injury. Circulation. 103:133-139, 2001
Yasuda SI, Sugiura S, Kobayakawa N, Fujita H,
Yamashita H, Katoh K, Saeki Y, Kaneko H, Suda Y,
Nagai R, Sugi H. A novel method to study contraction characteristics of a single cardiac myocyte using
carbon fibers. Am J Physiol Heart Circ Physiol.
281:H1442-6, 2001.
Ishizaka N, Aizawa T, Ohno M, Usui Si S, Mori I,
Tang SS, Ingelfinger JR, Kimura S, Nagai R.
Regulation and localization of HSP70 and HSP25 in
the kidney of rats undergoing long-term administration of angiotensin II. Hypertension. 39:122-128,
2002.

NO and intracellular signaling pathways
1. Nagata D, Suzuki E, Nishimatsu H, Yoshizumi M,
Mano T, Walsh K, Sata M, Kakoki M, Goto A, Omata
M, Hirata Y. Cyclin A downregulation and p21(cip1)
upregulation correlate with GATA- 6-induced growtharrest in glomerular mesangial cells. Circ Res. 87:
699-704, 2000.
2. Sata M, Kakoki M, Nagata D, Nishimatsu H, Suzuki
E, Aoyagi T, Sugiura S, Kojima H, Nagano T,
Kangawa K, Matsuo H, Omata M, Nagai R, Hirata Y.
Adrenomedullin and nitric oxide inhibit humanendothelial cell apoptosis via a cyclic GMP-independent mechanism. Hypertension. 36: 83-88, 2000.
3. Suzuki E, Nagata D, Yoshizumi M, Kakoki M, Goto
A, Omata M, Hirata Y. Reentry into the cell cycle ofcontact-inhibited vascular endothelial cells by aphosphatase inhibitor. Possible involvement of

4.

5.

6.

7.

extracellular signal-regulated kinase and phosphatidylinositol 3-kinase. J Biol Chem. 275: 36373644, 2000
Nagata D, Suzuki E, Nishimatsu H, Satonaka H, Goto
A, Omata M, Hirata Y. Transcriptional activation of
the cyclin D1 gene is mediated by multiple cis-elements, including SP1 sites and a cAMP-responsive
element in vascular endothelial cells. J Biol Chem.
276: 662-669, 2001.
Nishimatsu H, Suzuki E, Nagata D, Moriyama N,
Satonaka H, Walsh K, Sata M, Kangawa K, Matsuo
H, Goto A, Kitamura T, Hirata Y. Adrenomedullin
induces endothelium-dependent vasorelaxation via
the phosphatidylinositol 3-kinase/Akt-dependent
pathway in rat aorta.Circ Res. 89: 63-70, 2001.
Nishimatsu H, Hirata Y, Shindo T, Kurihara H,
Kakoki M, Nagata D, Hayakawa H, Satonaka H, Sata
M, Tojo A, Suzuki E, Kangawa K, Matsuo H,
Kitamura T, Nagai R. Role of endogenous
adrenomedullin in the regulation of vascular tone
and ischemic renal injury: studies ontransgenic/knockout mice of adrenomedullin gene.
Circ Res. 90: 657-663, 2002.
Suzuki N, Kojima H, Urano Y, Kikuchi K, Hirata Y,
Nagano T. Orthogonality of calcium concentration
and ability of 4,5- diaminofluorescein to detect NO. J
Biol Chem 277:47-49, 2002.

Nagai R. A novel biochemical diagnostic method for
aortic dissection - the results of a prospective study
using an immunoassay of smooth muscle myosin
heavy chain. Circulation 93:1244-1249, 1996
2. Yokoyama I, Yonekura K, Ohtake T, Yang W, Shin
WS, Yamada N, Ohtomo K, Nagai R. Coronary
microangiopathy in type 2 diabetic patients: relation
to glycemic control, sex, and microvascular angina
rather than to coronary artery disease. J Nucl Med
41: 978-985, 2000
3. Iwasaki T, Iwasaki T, Aihara Y, Kanda T, Oriuchi N,
Endo K, Katoh H, Suzuki T, Nagai R.
Immunoscintigraphy of aortic dissection with
99mTc-labeled murine anti-smooth muscle myosin
monoclonal antibody in rats. J Nucl Med 42: 130137, 2001
4. Yokoyama I, Yonekura K, Moritan T, Tateno M,
Momose T, Ohtomo K, Inoue Y, Nagai R.
Troglitazone improves whole-body insulin resistanceand skeletal muscle glucose use in type II diabetic
patients. J Nucl Med. 42:1005-1010, 2001.

Genetics of cardiovascular disease
1. Tanaka T, Nagai R, Tomoike H, Takata S, Yano K,
Yabuta K, Haneda N, Nakano O, Shibata A,
Sawayama T, Kasai H, Yazaki Y, Nakamura Y. Fournovel KVLQT1 and four novel HERG mutations infamilial long QT syndrome. Circulation 95: 565-567,
1997.
2. Nakajima T, Furukawa T, Tanaka T, Katayama Y,
Nagai R, Nakamura Y, Hiraoka M. Novel mechanismof HERG current suppression in LQT2 shift in voltage dependence of HERG inactivation. Circ Res
83:415-422, 1998.
3. Itoh T, Tanaka T, Nagai R, Kamiya T, Sawayama T,
Nakayama T, Tomoike H, Sakurada H, Yazaki Y,
Nakamura Y. Genomic organization and mutational
analysis of HERG, a gene responsible for familial long
QT syndrome. Hum Genet 102: 435-439, 1998
4. Itoh T, Tanaka T, Nagai R, Kikuchi K, Ogawa S,
Okada S, Yamagata S, Yano K, Yazaki Y, Nakamura
Y. Genomic organization and mutational analysis of
KVLQT1, a gene responsible for familial long QT
syndrome. Hum Genet 103: 290-294, 1998
5. Iwasa H, Itoh T, Nagai R, Nakamura Y, Tanaka T.
Twenty single nucleotide polymorphisms (SNPs) andtheir allelic frequencies in four genes that are responsible for familial long QT syndrome in the Japanese
population. J Hum Genet 45: 182-183, 2000
Diagnosis and imaging of cardiovascular disease
1. Suzuki T, Katoh H, Watanabe M, Kurabayashi M,
Hiramori K, Hori S, Nobuyoshi M, Tanaka H,
Kodama K, Sato H, Suzuki S, Tsuchio Y, Yazaki Y,

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Department of
Gastroenterology
Department Objectives
The goal of our department is to accurately diagnose and give the best available treatment to patients, and develop better modalities for the future.
To attain the goal, we perform clinical and basic research.

OutlineIn 1998, our current Department of Gastroenterology was established through the reorganization of the Divisions of Internal Medicine, and is in charge of busy clinical practices;
5,000 outpatients a month and 1,200 new admissions a year. The most frequent cause ofadmission is hepatocellular carcinoma, 500 a year, followed by gastrointestinal tract disorders 400, pancreatobiliary 180, and others 120.
Outline-Hepatology
Unlike other countries, the majority (more than 90%) of our patients with liver disease
are due to hepatitis virus infection. Therefore, better understanding of virus replication andpathogenesis may eventually lead to the eradication of the virus and the cure of diseases.
We have developed unique ablation treatment for hepatocellular carcinoma (HCC), forwhich many patients have visited our department (Fig. 1); we have had 1,600 cases, probably the largest series in the world as a single institute experience.
Outline -Gastroenterology
Gastric cancer is the commonest cause of death in Japan. Similar to HCC, there is now
enough evidence that bacterial infection, Helicobacter pylori, induces or is at least strongly
related to cancer development. Along with clinical expertise like en bloc resection of cancer(Fig. 2), better understanding of gastric injury due the infection could reduce the most common cause of death in Japan.
Recent increase in the number of colonic cancer is so noticeable. We found that this ismainly due to increase in right sided colon (cecum and ascending colon) cancer in elderly
patients. The better understanding of right sided as well as left sided colonic cancers may
prevent further increase in the colonic neoplasm in a country where people enjoy unprecedented length of life.
Outline -Pancreatobiliary
Pancreatic cancer with incredibly poor prognosis (average survival of 200 days) is insidiously increasing in Japan. There is not even a clue for high risk group. We need, though difficult, to find out high risk groups and better treatment.
Fig 1. Percutaneous tumor ablation.
(A) During treatment: fine needle was inserted into the tumor.
(B) After treatment: the tumor was completely ablated

Fig 2. Endoscopic tumor resection.
(A) After injection of saline, the tumor was cut around.
(B) en bloc resection: the tumor was completely removed.

164

Faculties and Students
Professor and Chair

Masao Omata, M.D., Ph.D. (from1992)
Lecturer
Takao Kawabe, M.D., Shin Onishi,
M.D.
Associate ...............................19
Postgraduate Fellow ............33
Graduate Student..................23
Research Studen .....................7
Secretary .................................2

Past Research and Major Accomplishments
1. Acute and Chronic Liver Injury due to HBV(Hepatitis B Virus)
The most severe form of acute liver injury is fulminant hepatitis of which 70% is fatal.
However, the underlying mechanisms of severe
injury have not been clear.
We elucidated for the first time the fulminant hepatitis was induced by HBV mutant form (N Engl J Med
1991;324:1699-1704).
Furthermore, this mutant carries specific aminoacid changes in nucleo-capsid region, a possible targetof lymphocyte attack (J Clin Invest 1993;91:1206 1213).
These mutations play a critical role not only inacute but also chronic liver disease (J Clin Invest 1992
;89:332-338, Gastroenterology 1993;104:263-271).
By these understandings, we explored how totreat these wild and mutant strains.
We set up in vitro system to maintain these wild
and mutant virus replication and identified extremely
effective drugs (J Clin Invest1999;103:1635-1640, J
Clin Invest 2001;107:449-455).
These drugs are now on clinical trials to prevent
fulminant hepatitis and even the decompensated cirrhosis to save patients life.
2. Liver Injury due to HCV (Hepatitis C Virus)
In contrast to HBV, HCV infection is mild but
insidious and long-lasting disease to carcinoma.
All of these start with very high chronicity rate(80%) of acute hepatitis. We elucidated for the firsttime that this is nearly 100% preventable by anti-viraltreatment(Lancet1991;338:914-915,
Gastroenterology 1994;107:805-811). Since HCV vaccine has not been developed, the above mentioned isthe best available procedure to prevent chronicity. In
fact, a study from Germany recently reported identicalresults (N Engl J Med 2001;345:1452-1457). However,there are stil1 2 million carriers who already havechronic infection. In fact, over 80% of patients with
HCC are due to HCV infection in Japan. To prevent

HCC, we first revealed natural course of chronic hepatitis to HCC ; this is by step-wise progression of hepatic fibrosis (Ann Intern Med 2000;132:517-524). And
eradication of virus by interferon could reduce theincidence of HCC (Ann Intern Med 1999;131:174-181)
with resolution of hepatic fibrosis even in cirrhosis,
previously thought irreversible (Ann Intern Med
2000;132:517-524). Furthermore, it was recently
revealed that overall mortality (not just HCC) was
reduced in treatment cohort (Gastroenterology
2002;123: 483-491). By these studies, we now know
how to prevent, but question is how to treat patients
who already have the cancer. Probably our department is the largest referral center of HCC (500 cases ayear), some are from outside Japan, seeking our medical tumor ablation treatment (Fig. 1).
We recently finished our 7-year prospective randomized trial in that we combined radical cure of thecancer nodules by ablation and eradication of HCV byinterferon. The 7-year survival of these patientsreached 75% (Ann Intern Med 2002; in press). This iscompatible or somewhat better than those reported inliver transplantation. When I graduated medical
school 30 years ago, liver disease was the one impossible to treat, but now able to envision to how to cureall.
3. Gastric Injury
Gastric cancer is number one cause of death inJapan. More than 80% is supposed to be due to H.
pylori infection; 90% eradication of the bacteria is now
possible by proper treatment. Thus, we decided toelucidate underlying mechanisms from infection to
the cancer, by in vitro and in vivo studies. First, we
tried to identify responsible molecules for inflammation.
In vitro
We revealed H. pylori induces strong
NF-kB signals, and these are by molecules in Cag
pathogenicity island, 40kb segment of 1.5 million
nucleotides (Gastroenterology 2000;119:97-108, J Biol
Chem 2001;276:44856-44864).
In vivo
To extrapolate this to in vivo system,
we constructed isogenic mutant (knock out bacteria),
lacking Cag gene molecules, and infected it to
Mongolian gerbils (unique animal model). It turned
out among Cag island, Cag E gene is the one responsible for strong induction of inflammation, whereas Vac
A, claimed to be inflammation inducing molecule, isnot (J Exp Med 2000;192:1601-1609). These CagE
molecule of H. pylori gene is generally present inJapanese isolates, whereas
it is often absent inEuropean (Gut 1998;42:338-343). These may explain
the difference of incidence of the gastric cancer
among different countries.
Recently, Kawakami and Tateishi further elucidated that IkBα, a key molecule in NF-kB signal cascade,165was regulated by NEDD8 pathway activating ubiquitin-proteasome degradation system (EMBO J
2001;20:4003-4012, J Cell Biol 2001;155:571-579).
Although the NF-kB signaling is extremely important as an inflammation inducing pathway, mechanism of direct effect on cell proliferation by H. pylori
is not known. Hirata recently elucidated that CagA
protein strongly activates SRE (Serum Response
Element) which might be operating in the formationof polyp and even MALT lymphoma(Gastroenterology 2002; in press).
Our search for effector protein to induce signalingcascades has now began (J Exp Med 2000;191:593602); it may help to eventually elucidate the molecularmechanism of gastric cancer.
4. Colon Cancer
Colonic cancer is catching up with the leading gastric cancer. Shift from Japanese to “Westernized” food
may be responsible for this increase. However, we
believe there is another reason. We have noticed rightsided (cecum and ascending) colon cancer is increasing among patients over age 70 (60% vs 20% in ages
over 70 and under 40, respectively) (Gastrointest
Endosc 2002;55:548-551). We found the majority of
the right sided colon cancer carried microsatellite
instability (Cancer Res 1996;56:5620-5623). Now inJapan, average life expectancy for female and male is
85 and 78, respectively, longest ever.
We have initiated genome-wild screening of thegenes having dinucleotide repeat or poly-A stretches,targets of repair gene errors.
5. Pancreatic Cancer
Once pancreatic cancer developed, median survival
was only 200 days. Although we tried to employ K-ras
gene mutation, which is found in 90% of the cancer,
for early diagnosis (Gastroenterology 1991;100:233238, 1996;110:227-231), it has been far from giving
benefit to patients.
We have struggled to find better way of treatment
and finally decided to apply allogenic stem cell transplantation by collaboration with Department of
Hematology and Oncology. So far 5 patients enrolled
and results are pending.

Current Research and Clinical Activities
1. Eradication of HBVAs I wrote, the ultimate goal of HBV treatment is
not mere suppression which is currently available, but
eradication of the virus (Treatment of chronic hepatitis B infection -Editorial N Engl J Med 1998 ; 339:114115). To attain this goal we are undertaking the following :166

HBV has two replication pathways, episomal (NEngl J Med 1986;315:1187-1192) and integrated form
into human genome which presumably resist thetreatment. We are now intensively focusing our study
on the integrated form by use of post-sequencing
information on human genome.
2. Eradication of HCV
We could eradicate the virus in more than 90% of
acute hepatitis cases as was shown by us (Lancet
1991;338:914-915). However, only 30% in chronic
hepatitis. To increase treatment efficacy, we now have
Replicon system which allows us to grow treatmentresistant strains and to test drug candidates.
3. Liver Injury Mechanisms
Sequencing of extreme small amount of peptides isnow feasible by high quality mass spectrometry. We
are now analyzing HLA-related wild and mutants
virus-derived peptides from human liver. This may
identify the target of lymphocyte attack.
4. Prevention and Cure of HCC
We have treated 1,600 cases of HCC so far. We
have shown that treatment of virus infection could
prevent HCC. However, advanced HCC is still difficult
to treat. We found that Vitamin K could efficiently
prevent portal tumor invasion, once developed 100%
fatal in one year. Along with other modalities, wemight expect long-term survival in the majority of thepatients with HCC.
5. Gastric Injury Mechanisms
Discovery of H. pylori protein in host cell, phosphorylated CagA protein (J Exp Med 2000;191:593602), has prompted us to further search the effector
protein, which could trigger signaling cascades. We
have now several candidates.
6. Right-sided Colon Carcinogenesis
As I mentioned, repair-gene system seems deeply
involved in the right-sided colon cancer, a prototype
is, of course, HNPCC (Hereditary Nonpolyposis Colon
Cancer). The understanding may help for people inthe country where longest life-expectancy expected.
7. Pancreatic Cancer
We have initiated a large scale genome-wide SNP(Single Nucleotide Polymorphism) analysis to findhigh risk group.

Others Activites
In 5 years, we presented 121 papers in DDW(Digestive Disease Week, USA), the largest gastroenterology congress.

In 5 years, we published 320 papers in peer review
journal.
In 5 years, 52 invited lectures were given in international meetings by one of us (M Omata).

Future Prospective
Education for medical students and for young
physicians is the far most important mission of ourdepartment for the future. Our department have tocover multiple organs with diverse diseases. Thus, weare trying not to restrict their interest to one specialized field, but rather to introduce general technology
and ideas. It will definitely become more “borderless”
in post-genome era.

Research Grants
The Organization for Pharmaceutical Safety
Research 1997-2001 (Total ￥346,000,000.)
Comprehensive 10-year Strategy for Cancer
Control from Ministry of Health and Welfare, Health
Sciences Research Grants 1994-1999 (Total￥100,600,000.)
Medical Frontier Strategy Research from Ministryof Health Labor and Welfare, Health Sciences
Research Grants 2001-2003 (2001 / ￥48,000,000.)
Grants-in-aid for Scientific Research on Priority
Area from the Ministry of Education, Culture, Sports,
Science and Technology of Japan 2000-2003 (Total￥71,800,000.)
Grants-in-aid for Scientific Research (A) from the
Ministry of Education, Culture, Sports, Science andTechnology of Japan 2001-2004 (Total ￥40,300,000.)

Select Publications (50)
Hepatology
1. Omata M, Ehata T, Yokosuka O, Hosoda K, Ohto M.
Mutations in the precore region of hepatitis B virus
DNA in patients with fulminant and severe hepatitis.
N Engl J Med 324 : 1699-1704, 1991.
2. Omata M, Yokosuka O, Takano S, Kato N, Hosoda K,
Imazeki F, Tada M, Ito Y, Ohto M. Resolution of
acute hepatitis C after therapy with natural beta
interferon. Lancet 338 : 914-915, 1991.
3. Ehata T, Omata M, Yokosuka O, Hosoda K, Ohto M.
Variations in codon 84-101 in the core nucleotide
sequence correlate with hepatocellular injury inchronic hepatitis B virus infection. J Clin Invest 89 :332-338, 1992.
4. Hosoda K, Omata M, Yokosuka O, Kato N, Ohto M.
Nucleotide sequences of 5- 1-1 of hepatitis C virus inpatients with chronic liver disease. Gastroenterology
102 :1039-1043, 1992.

5. Chien DY, Choo QL, Tabrizi A, Kuo C, McFarland J,
Berger K, Lee C, Shuster JR, Nguyen T, Moyer DL,
Tong M, Furuta S, Omata M, Tegtmeier G, Alter H,
Schiff E, Jeffers L, Houghton M, Kuo G. Diagnosis ofhepatitis C virus (HCV) infection using an immunodominant chimeric polyprotein to capture circulating antibodies : reevaluation of the role of HCV inliver disease. Proc Natl Acad Sci USA 89 : 1001110015, 1992.
6. Ehata T, Omata M, Chuang WL, Yokosuka O, Ito Y,
Hosoda K, Ohto M. Mutations in core nucleotide
sequence of hepatitis B virus correlate with fulminant and severe hepatitis. J Clin Invest 91 : 12061213, 1993.
7. Chuang WL, Omata M, Ehata T, Yokosuka O, Ito Y,
Imazeki F, Lu SN, Chang WY, Ohto M. Precore mutations and core clustering mutations in chronic hepatitis B virus infection. Gastroenterology 104 : 263271, 1993.
8. Takano S, Satomura Y, Omata M, Japan Acute
Hepatitis Cooperative Study Group. Effects of interferon beta on Non-A, Non-B acute hepatitis: A
prospective, randomized, controlled-dose study.
Gastroenterology 107 : 805-811, 1994.
9. Takano S, Yokosuka O, Imazeki F, Tagawa M, Omata
M. Incidence of hepatocellular carcinoma in chronic
hepatitis B and C : A prospective study of 251
patients. Hepatology 21 : 650-655, 1995.
10. Matsumoto A, Ogura K, Hirata Y, Kakoki M,
Watanabe F, Takenaka K, Shiratori Y, Momomura S,
Omata M. Increased nitric oxide in the exhaled air ofpatients with decompensated liver cirrhosis. Ann
Intern Med 123 : 110-113, 1995.
11. Shiratori Y, Shiina S, Imamura M, Kato N, Kanai F,
Okudaira T, Teratani T, Tohgo G, Toda N, Ohashi M,
Ogura K, Niwa Y, Kawabe T, Omata M.
Characteristic difference of hepatocellular carcinoma
between hepatitis-B and -C viral infection in Japan.
Hepatology 22 : 1027-1033, 1995.
12. Kanai F, Lan KH., Shiratori Y, Tanaka T, Ohashi M,
Okudaira T, Yoshida Y, Wakimoto H, Hamada H,
Nakabayashi H, Tamaoki T, Omata M. In vivo genetherapy for alpha-fetoprotein-producing hepatocellular carcinoma by adenovirus-mediated transfer ofcytosine deaminase gene. Cancer Res 57 : 461-465,
1997.
13. Shiratori Y, Kato N, Yokosuka O, Imazeki F,
Hashimoto E, Hayashi N, Nakamura A, Asada M,
Kuroda H, Tanaka N, Arakawa Y, Omata M.
Predictors of the efficacy of interferon therapy inchronic hepatitis C virus infection. Gastroenterology
113 : 558-566, 1997.
14. Kojima H, Yokosuka O, Imazeki F, Saisho H, Omata
M. Telomerase activity and telomere length in hepatocellular carcinoma and chronic liver disease.
Gastroenterology 112 : 493-500, 1997.
15. Omata M. Treatment of chronic hepatitis B infection(Editorial). N Engl J Med 339 : 114-115, 1998.
16. Ono-Nita SK, Kato N, Shiratori Y, Lan KH, Yoshida
H, Carrilho FJ, Omata M. Susceptibility of lamivu-

167

dine-resistant hepatitis B virus to other reverse transcriptase inhibitors. J Clin Invest 103 : 1635-1640,
1999.
17. Yoshida H, Shiratori Y, Moriyama M, Arakawa Y, Ide
T, Sata M, Inoue O, Yano M, Tanaka M, Fujiyama S,
Nishiguchi S, Kuroki T, Imazeki F, Yokosuka O,
Kinoyama S, Yamada G, Omata M. Interferon therapy reduces the risk for hepatocellular carcinoma :National surveillance program of cirrhotic and noncirrhotic patients with chronic hepatitis C in Japan.
Ann Intern Med 131 : 174-181, 1999.
18. Shiratori Y, Imazeki F, Moriyama M, Yano M,
Arakawa Y, Yokosuka O, Kuroki T, Nishiguchi S,
Sata M, Yamada G, Fujiyama S, Yoshida H, Omata
M. Histologic improvement of fibrosis in patientswith hepatitis C who have sustained response tointerferon therapy. Ann Intern Med 132 : 517-524,
2000.
19. Kato N, Yoshida H, Ono-Nita SK, Kato J, Goto T,
Otsuka M, Lan KH, Matsushima K, Shiratori Y,
Omata M. Activation of intracellular signaling byhepatitis B and C viruses: C-viral core is the mostpotent signal inducer. Hepatology 32 : 405-412,
2000.
20. Otsuka M, Kato N, Lan KH, Yoshida H, Kato J, Goto
T, Shiratori Y, Omata M. Hepatitis C virus core protein enhances p53 function through augmentation of
DNA binding affinity and transcriptional ability. J
Biol Chem 275 :34122-34130, 2000.
21. Goto T, Kato N, Ono-Nita SK, Yoshida H, Otsuka M,
Shiratori Y, Omata M. Large isoform of hepatitis
delta antigen activates serum response factor-associated transcription. J Biol Chem 275 : 37311-37316,
2000.
22. Koike Y, Shiratori Y, Sato S, Obi S, Teratani T,
Imamura M, Yoshida H, Shiina S, Omata M. Des-rcarboxy prothrombin as a useful predisposing factor
for the development of portal venous invasion inpatients hepatocellular carcinoma -a prospective
analysis of 277 patients-. Cancer 91 : 561-569, 2001.
23. Ono-Nita SK, Kato N, Shiratori Y, Kato J, Goto T,
Schinazi RF, Carrilho FJ, Omata M. The polymerase
L528M mutation cooperates with nucleotide binding-site mutations, increasing hepatitis B virus replication and drug resistance. J Clin Invest 107 : 449455, 2001.
24. Yoshida H, Kato N, Shiratori Y, Otsuka M, Maeda S,
Kato J, Omata M. Hepatitis C virus corprotein activates nuclear factor κB-dependent signaling throughtumor necrosis factor receptor-associated factor. J
Biol Chem 276 : 16399-16405, 2001.
25. Yoshida H, Arakawa Y, Sata M, Nishiguchi S, Yano
M, Fujiyama S, Yamada G, Yokosuka O, Shiratori Y,
Omata M. Interferon therapy prolonged life
expectancy among chronic hepatitis C patients.
Gastroenterology 123 : 483-491, 2002.
26. Lan KH, Sheu ML, Hwang SJ, Yen SH, Chen SY, Wu
JC, Wang YJ, Kato N, Omata M, Chang FY, Lee SD.
HCV NS5A interacts with p53 and inhibits p53mediated apoptosis. Oncogene 21 : 4801-4811, 2002.

168

27. Shiratori Y, Shiina S, Teratani T, Imamura M, Obi S,
Sato S, Koike Y, Yoshida H, Omata M. Interferon
therapy after tumor ablation improves prognosis inpatients with hepatitis C viral hepatocellular carcinoma. Ann Inten Med 2002 (in press).

Gatroenterology
28. Tada M, Omata M, Ohto M. Clinical application of
ras-gene mutation for diagnosis of pancreatic adenocarcinoma. Gastroenterology 100 : 233-238, 1991.
29. Tada M, Omata M, Kawai S, Saisho H, Ohto M, Saiki
R, Sninsky J. Detection of ras gene mutations in pancreatic juice and peripheral blood of patients withpancreatic adenocarcinoma. Cancer Res 53 : 24722474, 1993
30. Ohta S, Yoshiura K, Takahashi M, Hata Y, Kohmoto
O, Kawabe T, Shimada T, Hiraishi H, Mutoh H,
Terano A, Sugimoto T, Omata M. P2 purinergic
receptor regulation of mucus glycoprotein by rabbit
gastric mucous cells in a primary culture.
Gastroenterology 106 : 1485-1492, 1994.
31. Shimada T, Ogura K, Ota S, Terano A, Takahashi M,
Hamada E, Omata M, Sumino S, Sassa R.
Identification of helicobacter pylori in gastric specimens, gastric juice, saliva and faeces of Japanesepatients. Lancet 343 : 1636-1637, 1994.
32. Takahashi M, Ota S, Shimada T, Hamada E, Kawabe
T, Okudaira T, Matsumura M, Kaneko N, Terano A,
Nakamura T, Omata M. Hepatocyte growth factor isthe most potent endogenous stimulant of rabbit gastric epithelial cell proliferation and migration in primary culture. J Clin Invest 95 : 1994-2003, 1995.
33. Tada M, Ohashi M, Shiratori Y, Okudaira T, Komatsu
Y, Kawabe T, Yoshida H, Machinami R, Kishi K,
Omata M. Analysis of K-ras gene mutation in hyperplastic duct cells of the pancreas without pancreaticdisease. Gastroenterology 110 : 227-231, 1996.
34. Imazeki F, Yokosuka O, Yamaguchi T, Ohto M, Isono
K, Omata M. Expression of variant CD44-messenger
RNA in colorectal adenocarcinomas and adenomatous polyps in humans. Gastroenterology 110 : 362368, 1996.
35. Lan KH, Kanai F, Shiratori Y, Okabe S, Yoshida Y,
Wakimoto H, Hamada H, Tanaka T, Ohashi M,
Omata M. Tumor-specific gene expression in carcinoembryonic antigen-producing gastric cancer cells
using adenovirus vectors. Gastroenterology 111 :
1241-1251, 1996.
36. Togo G, Toda N, Kanai F, Kato N, Shiratori Y, Kishi
K, Imazeki F, Makuuchi M, Omata M. A transforming growth factor beta type II receptor gene mutationcommon in sporadic cecum cancer with microsatellite instability. Cancer Res 56 : 5620-5623, 1996.
37. Takahashi M, Ota S, Hata Y, Mikami Y, Azuma N,
Nakamura T, Terano A, Omata M. Hepatocyte
growth factor as a key to modulate anti-ulcer actionof prostaglandins in stomach. J Clin Invest 98 : 26042611, 1996.
38. Lan KH, Kanai F, Shiratori Y, Ohashi M, Tanaka T,
Okudaira T, Yoshida Y, Hamada H, Omata M. In

vivo selective gene expression and therapy mediatedby adenoviral vectors for human carcinoembryonicantigen-producing gastric carcinoma. Cancer Res 57 :4279-4284, 1997.
39. Maeda S, Ogura K, Yoshida H, Kanai F, Ikenoue T,
Kato N, Shiratori Y, Omata M. Major virulence factors, VacA and CagA, are commonly positive in
Helicobacter pylori isolates in Japan. Gut 42 : 338343, 1998.
40. Maeda S, Yoshida H, Ikenoue T, Ogura K, Kanai F,
Kato N, Shiratori Y, Omata M. Structure of cag pathogenicity island in Japanese Helicobacter pylori isolates. Gut 44 : 336-341, 1999.
41. Asahi M, Azuma T, Ito S, Ito Y, Suto H, Nagai Y,
Tsubokawa M, Tohyama Y, Maeda S, Omata M,
Suzuki T, Sasakawa C. Helicobacter pylori Cag A protein can be tyrosine phosphorylated in gastric epithelial cells. J Exp Med 191 : 593-602, 2000.
42. Maeda S, Yoshida H, Ogura K, Mitsuno Y, Hirata Y,
Yamaji Y, Akanuma M, Shiratori Y, Omata M. H.
pylori activates NF-κB through a signaling pathway
involving IκB kinases, NF-κB-inducing kinase,
TRAF2, and TRAF6 in gastric cancer cells.
Gastroenterology 119 : 97-108, 2000.
43. Ogura K, Maeda S, Nakao M, Watanabe T, Tada M,
Kyutoku T, Yoshida H, Shiratori Y, Omata M.
Virulence factors of Helicobacter pylori responsible
for gastric diseases in Mongolian gerbil. J Exp Med
192 : 1601-1609, 2000.
44. Kawakami T, Chiba T, Suzuki T, Iwai K, Yamanaka
K, Minato N, Suzuki H, Shimbara N, Hidaka Y,
Osaka F, Omata M, Tanaka K. NEDD8 recruits E2ubiquitin to SCF E3 ligase. EMBO J 20 : 4003-4012,
2001.
45. Matsuhashi N, Watanabe K, Kanamori H, Ohnishi S,
Omata M. PPARgamma agonists for intestinal
ischaemia. Lancet 358 :75-76, 2001.
46. Mitsuno Y, Yoshida H, Maeda S, Ogura K, Hirata Y,
Kawabe T, Shiratori Y, Omata M. Helicobacter pylori
induced transactivation of SRE and AP-1 through the
ERK signalling pathway in gastric cancer cells. Gut 49:18-22, 2001.
47. Tateishi K, Omata M, Tanaka K, Chiba T. The
NEDD8 system is essential for cell cycle progression
and morphogenetic pathway in mice. J Cell Biol 155 :571-579, 2001.
48. Maeda S, Akanuma M, Mitsuno Y, Hirata Y, Ogura K,
Yoshida H, Shiratori Y, Omata M. Distinct mechanism of Helicobacter pylori-mediated NF-kappa B
activation between gastric cancer cells and monocytic cells. J Biol Chem 276 : 44856-44864, 2001.
49. Okamoto M, Shiratori Y, Yamaji Y, Kato J, Ikenoue T,
Togo G, Yoshida H, Kawabe T, Omata
M.Relationship between age and site of colorectal
cancer based on colonoscopy findings. Gastrointest
Endosc 55 : 548-551, 2002.
50. Hirata Y, Maeda S, Mitsuno Y, Akanuma M, Yanai A,
Yoshida H, Kawabe T, Shiratori Y, Omata M. H.
pylori CagA protein activates serum response ele-ment driven transcription independently of tyrosine
phosphorylation.Gastroenterology 2002 (in press).

169

Department of Nephrology
and Endocrinology
Outline and Research Objectives
The Department of Nephrology and Endocrinology is one of the major divisions in the
Department of Internal Medicine the University of Tokyo. It covers nephrology, endocrinology, and hypertension. In 1998, the Department of Internal Medicine of the University ofTokyo has been reorganized. Since then, Dr. Fujita has been Professor and Chairman of ourdepartment. His achievements touch on hypertension and sodium metabolism. This is oneof our main research objectives. The professor and each member of staff have an active
responsibility for all research, clinical, and educational activities. There are 12 AssistantProfessors supporting the clinical works of our residents in the ward, and also involved in
the Hemodialysis Unit. We are working side-by-side in all clinical activities under the supervision of the professor.
In our department, renal biopsy is actively performed and the morphrogical diagnosis of
the nephritis is established to give the real benefits of treatment to the patients. We also
treat diabetic patients with proteinuria and end-stage renal failure. Every staff physician ofour division also works at the hemodialysis unit thus we can manage patients in every stageof renal disease. The endocrine subunit attends a broad spectrum of endocrine entities providing the residents and academic internists a high quality of medical training. It is also our
speciality to diagnose and treat secondary hypertension caused by primary aldosteronism,
Cushing’s syndrome, pheochromocytoma and so on. We often have consultations from theother divisions concerning water and mineral metabolism disorders.
In our division, there are more than 30 students of the Graduate School. This suggests
high research activities and outstanding leadership of individual staff in our division.
Research seminars are held every Tuesday evening, where both senior investigators andtrainees present their own data and experimental plans and discuss profoundly with theprofessor and all members of our division. In the seminars, scientists outside our department and outside the University of Tokyo including foreign countries sometimes lecture
their papers, which encourages our research activities. Moreover, we are actively collaborating with some of the scientists outside our department. Our research topics are covered thealmost all field of hypertension, nephrology, and endocrinology.

Faculties and Students
Professor and Chair Toshiro Fujita. M.D., Ph.D.
Associate Professors Atsuo Goto, M.D., Ph.D.
Tomoki Okazaki, M.D., Ph.D.
Lecturers
Shigeo Taniguchi, M.D., Ph.D.
Kenmei Takaichi, M.D., Ph.D.
Assistant Professors ...............6 nephrologists and 6
endocrinologists
Post-doctorial fellows .............9
Graduate Students ................38
Research Students...................2
Secretaries ...............................2

Past Research and Major Accomplishments
The following major accomplishment has beenobtained using the best available molecular, physiological, and morphological techniques as possible.

170

Vasoactive substances play an important role in
not only blood-pressure control but also formation ofvascular damages. Thus, we focused on
adrenomedullin (AM), a novel vasodilator peptide. To
elucidate physiological and pathophysiological function of AM, we generated AM knockout mice.
Homozygous AM knockout (AM-/-) mice were embryonic lethal but heterozygous (AM+/-) mice were grewnormally although its AM levels in plasma and tissues
were as a half as wild-type mice. Angiotensin II (AII)and salt loading caused severe coronary arterial damages in AM +/- mice. Moreover, AM +/- mice showedenhancement of cuff injury-induced intimal hyperplasia of femoral artery. These changes in AM+/- mice
were accompanied with enhanced oxidative stress,
suggesting that antioxidant effect of AM is importantin its protective effect against vascular damages. Thus,
intrinsic AM counteracts AII, and exhibits vasoprotective action.

Insulin resistance has been suggested to play animportant role in hypertension and the related organ
damages. Thus, we also focused on insulin resistancein several model animals of hypertension.
Interestingly, salt loading itself enhanced insulin
resistance in normotensive rats. Certainly, saltinduced rise in blood pressure in Dahl salt-sensitive
and angiotensin II (AII)-treated rats was accompaniedwith enhanced insulin resistance. The increased
insulin resistance in hypertensive models is intimately related to oxdative stress. Interestingly, aged AM
deficient mice showed insulin resistance, associatedwith overproduction of oxidative stress. Then, this is
the model of insulin resistance syndrome, so-called“syndrome X”.
Oxidized low density lipoprotein (OxLDL) has
been implicated in pathophysiology of atherosclerosis.
OxLDL alters endothelial function and triggered
atherogenic changes. Thus, we focused on a novel
endothelial OxLDL receptor, lectin-like OxLDL receptor-1 (LOX-1). We observed that LOX-1 was enhanced
in aortas and kidneys of rats with hypertension, diabetes, and renal failure. Also, LOX-1 was stimulatedby oxdative stress, which is known to cause organ
damages by common diseases. The expression of
LOX-1 was intimately related to nuclear factor (NF)kB. Thus, LOX-1 was implicated in pathphysiology ofatherosclerosis and glomeruloscrelosis via oxidative
stress.
To elucidate a novel candidate gene for salt sensitiviy of blood pressure, we performed differential display assay between Dahl salt-sensitive (DS) and–resistant (DR) rats. As a result, we found REPT1 (ratretrovirus-like element transcribed in proximal tubulus) in the kidneys of DS rats. REPT1 was locatd in
the proximal tubules and its renal expression was correlated with salt sensitivity among different strains ofrats.
Intracellular calcium ion (Ca2+) is a spatially andtemporally organized second messenger, which mediates a variety of important functions of vascularendothelial cells ranging from the production ofvasoactive substances to gene expression. We developed FRET-based technique to image localized Ca2+
signaling in live endothelial cells. And, we succeeded
to elucidate localized Ca2+ signaling and function of
caveolae. Endothelial nitric oxide (NO) was localizedin caveolae and activity of NO was regulated by intracellular Ca2+ and binding to caveolae.
G proteins control a diverse network of signals.
We have focused on mutation of G proteins in humandiseases. We have discovered how point mutations inaS (the a subunits of GS) alter GS signaling, causing
human diseases. Also, we succeeded in building up a
simple cell-free method to study interaction between
receptors and G proteins. We have, for the first time,achieved success in designing dominant negative
mutant Gα proteins in realistic level. Also, we analyzed relationship of C825T polymorphism of Gβ3 tohypertension, which has been demonstrated in
Caucasian people.
We revealed that ionic mechanisms such as activation of inwardly-rectifying potassium currents andactivation of nonselective cation currents are important in the regulation of hormone secretion from pituitary cells using electrophysiological techniques. Also,
we elucidated the mechanisms of action of regulatorsof circulation system such as AM, proadrenomedullin
N-terminal 20 peptide (PAMP), and AII. In addition,
we studied mineralocorticoid action in pathophysiology of cardiovascular diseases.
We studied molecular basis for diseases of calcium
2+(Ca ) metabolism. For example, we have identified
novel mutations in Ca-sensing receptor gene inpatients with Type ⅡBartter ‘s syndrome as well as
those with familial hypocalciuric hypercalcemia(FHH). Also, we characterized proteoglycans synthesized by osteoblasts biochemically and explored indetail cell-matrix interactions in osteoblasts. In addition, we identified negative Ca2+ responsive element(nCaRE) in the parathyroid hormone (PTH) gene.
From a physiological standpoint, we elucidated thebasic transport properties of sodium-bicarbonate (Na+HCO3-) cotransport (NBC1) in renal proximal tubularacidosis (RTA) with ocular abnormalities. This is an
example of a successful translation of basic finding
into clinical medicine. We clarified the regulatory
mechanism of proximal transport in physiological andpathological states and demonstrated the cAMP-activated chloride (Cl-) conductance in proximal tubules.
We revealed the immunohistochemical localizationand function of Kir 7.1 potassium channel, organic
anion transporter (OAT1), and stanniocalcin in the rat
kidney. We cloned the Na + /H + antiporter (NHE)
cDNAs from human tissues and characterized the regulation of the expression of NHE in kidney cells.
With the application to acute renal failure or graft
survival in mind, we demonstrated the successfulintervention by cyclic RGD (Arg-Gly-Asp) peptide to
reduce the renal ischemia-reperfusion model and the
diagnistic utilities of monitoring the radio-labbeled
RGD peptides. We demonstrated the incorporation ofoligonucleotides to proximal tubular epithelial cells
and the amelioration of renal ischemia-reperfusion
injury using antisense oligonucleotides specificiallytargeted to inducible NO synthase (NOS). We showedby in vivo approach the chemotactic activity ofleukotriene B4 to drive neutrophil in acute ischemiareperfusion injury.
The clarification of the mechanism of the progressive renal disease is the major problem to be solved in
the field of nephrology. We contribute much to the171solution of immunological mechanism involved inglomerulonephritis, especially in terms of complement and complement regulatory proteins. We
revealed the localization of NOS and NAD(P)H oxidase isoforms in the kidney and gave insight into thefunctional role of NO and oxidative stress in the kidney of hypertension, glomerulonephritis, and diabetes. We examined the role of physical factors suchas mechanical stretch on TGF-β and extracellularmatrix accumulation in the progression of renal disease. We demonstrated that renoprotective C-typenatriuretic peptide was produced in renal parenchymal cells.
We isolated several transcription factors from vascular myocytes such as an isoform of MEF2A and
GATA-6 and identified the implication of these factors
in the differentiation or dedifferentiation of vascular
myocytes and vascular hypertrophy process. We clarified the intracellular pathway involved in the cell
cycle progression with endothelin-1 proved to promote the renal injury.
As a new technique, we developed a simple andreliable method of quantitating mRNAs in minute tissue samples in addition to the established microdissection and single transduction mechanism by whichvasopressin and prostaglandins interact in the kidneyusing immunodissected tubule. We also developed anovel animal model of glomerular endothelial injury,
which may help elucidate the mechanism of glomerular endothelial damage.
Research on renal regeneration has just started to
do the identification of adult stem cells in the kidney.
Using a flow cytometry assay and Hoechst 33342
staining, we isolated adult stem cells in the kidney asside population (SP) cells that have been identified asstem cells in bone marrow, skeletal muscle and liver.
Using 3 dimensional culture systems, moreover, cultured renal SP cells differentiate into various kinds ofcells.
Troughout our research we have attempted to link
our basic finding with our clinical work-----between
bench and bedside with either direction.

Current Research
To extent our past accomplishment further, we are
now attempting to clarify the following themes.
Recent studies have revealed that oxidative stressis intimately related to not only hypertension but also
the other common disease such as diabetes andhypercholesterolemia, so we are examining whether
these abnormalities including insulin resistance develop AM+/- mice, in which oxidative stress is enhanced.
Also, we are investigating the effect of hypoxia in
AM +/- mice, because hypoxia-induced damage isknown to relate to oxidative stress.

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To elucidate atherogenic role of LOX-1, we are
studying whether atherogenic stimuli such as AII
infusion and cuff injury cause severe vascular damages in LOX-1 transgenic mice. Moreover, we foundthat peroxisome proliferator-activated receptor γ(PPARγ) ligands inhibit cytokine-stimulated LOX-1
expression in vitro and in vivo. Now we are studying
this mechanism, which may suggest that the inhibition of LOX-1 might be related to antiatherogenic
effect of PPARγ ligands.
We developed FRET-based tool to image subplasmalemmal Ca2+ and cAMP. We are now studying roleof caveolin scaffolding domain and hsp90 on NO production using unique cell biological and imaging techniques.
By the cell-free method to study interaction
between receptors and G proteins, we studied thespecificity between many G proteins and G proteincoupled receptors for cardiovascular hormones, suchas AII, catecholamines, and AM.
We are studying the central role of Ca-sensing
receptor in salt-sensitive hypertensive model animal,
in which Ca loading ameliorated hypertension.
Moreover, we have many themes such as electrophysiology of pituitary, signal transduction of mineralocorticoid receptor, mechanisms of age-related osteopenia,
and so on.
We further clarify molecular mechanism of proximal RTA based on newly identified NBC1 mutation.
The molecular basis of ion transport in NBC1 is studying via giant patch technique.
We are studying roles of lipid mediators in renal
ischemia-reperfusion injury using several lines of micedeleted/overexpressed lipid mediator genes. We are
analyzing detailed intracellular pathways implicatedin the expression of monocyte chemoattractant protein-1 in vascular myocytes.
We are investigating role of endogenous vasoactive
substances in the progression of renal diseases suchas AII, AM, leukotriene, retinoid and glucocorticoids
using knockout mice and renal cells. We are also
studying roles of oxidative and nitrosoactive stress in
the renal diseases including diabetic nephropathy,
ischemia-reperfusion injury, and long-term hemodialysis complications.
Now 3 projects of renal regeneration are on going.
First, optimizing the 3 dimensional culture systems todifferentiate renal SP cells to metanephros. Second,clarifying the condition and identifying the morphogen to differentiate human adult stem cells suchas bone marrow derived mesenchymal stem cells into
renal components. Third, investigating the existenceof stem cell dysfunction in patients with renal dysfunction, and getting comprehensive information forintervention using DNA microarray analysis.

Future Prospects
The main goal of our studies is to cure patientswith renal and endocrinological diseases and hypertension to reduce the number of patients with hypertensive complications and renal failure. To serve thispurpose, we have to focus our effort to clarify themechanisms of diseases. There are not only numerous
diseases in our field but also multiple mechanisms to
involved especially in chronic stage of renal failureand caidiovascular complication-associated hypertension. Thus, we are applying the multi-dimentional
approach such as oxidative stress, lipid metabolism,
hemodynamics, and vasoactive substabces. These
approaches will lead to the development of several
new drugs and strategies to treat renal and endocrinological diseases, hypertension, and its complications.
By the discoveries of the candidate genes for salt-sensitive hypertension, we can try to choose the suitableantihypertensive therapy and salt restriction for individual patients (tailor-made medicine).

Research Grants
1. Toshiro Fujita: 1998-2002 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (specificfield): Molecular biological and embryological investigation of adrenomedullin, 54,5000,000 yen
2. Toshiro Fujita: 2000-2002 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (B) Role ofoxidized LDL/lectin-like oxidized LDL receptor-1(LOX-1) system in the development of glomerulosclerosis. 15,000,000 yen
3. Atsuo Goto: 1995-1997 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (B) Role ofendogenous digitalis-like substance in relationshipbetween stress and cardiovascular diseases,
7,500,000 yen
4. Tomoki Okazaki: 1999-2001 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (B): The
mechanism of transcriptional inhibition by nuclear
hormone receptors by using a viamine D receptor asa model, 14,000,000 yen
5. Taro Iiri (Assistant professor of endocrinology): 20022004 Grant-in-aid for scientific research from the
Ministry of Education, Culture, Sports, Science andTechnology of Japan (A): G protein and diseases:
Analysis of molecular mechanism and regulation of
signal transduction, 30,000,000 yen

Select Publications
1. Watanabe S, Fukumoto S,Chang H, Takeuchi Y,
Hasegawa Y, Okazaki R, Chikatsu N,Fujita T:

Association between activating mutations of calcium-sensing receptor and Bartter’s syndrome.
Lancet,360:692-694,2002
2. Ogihara T, Asano T, Ando K, Sakoda H, Anai M.
Shojima N, Ono H, Onishi Y, Fujimoto M, Abe M.
Fukushima Y, Kikuchi M, Fujita T: High-Salt Diet
Enhances Insulin Signaling and Induces Insulin
Resistanse in Dahl Salt-Sensitive Rats, Hypertension,
2002;40:83-89.
3. Shimosawa T. Shibagaki Y. Ishibashi K. Kitamura K.
Kangawa K. Kato S. Ando K. Fujita
T:.Adrenomedullin, an endogenous peptide, counteracts cardiovascular damage. Circulation. 105(1):10611, 2002.
4. Sun D. Seki G. Uwatoko S. Nakao A. Goto A. Fujita T.
Kimura S. Taniguchi S: Quantifying porphobilinogendeaminase mRNA in microdissected nephron segments by a modified RT-PCR. Kidney International.
61(1):336-41, 2002.
5. Onozato ML. Tojo A. Goto A. Fujita T:. Wilcox CS:
Oxidative stress and nitric oxide synthase in rat diabetic nephropathy: effects of ACEI and ARB.Kidney
International. 61(1):186-94, 2002.
6. Chabrashvili T, Tojo A, Onozato M, Kitiyakara C,
Quinn M, Fujita T: Welch W, Wilcox S : Expression
and cellular localization of classic NADPH oxidasesubunits in the spontaneously hypertensive rat kidney. Hypertension 39:269-274,2002.
7. Suzuki E, Nishimatsu H, Satonaka H, Walsh K, Goto
A, Omata M, Fujita T: Angiotensin Ⅱ induces
myocyte enhancer factor 2- and calcineurin/nuclear
factor of activated T call-dependent transcriptional
activation in vascular myocytes. Circ Res. 90:10041011,2002.
8. Usui T, Hara M, Satho H, Moriyama N, Kagaya H,
Amano S, Oshita T, Ibaraki N, Hara C, Kunimi M,
Norii E, Tsukamoto K, Inatomi J, Kawakami H,
Endou H, Igarashi T, Goto A, Fijita T, Araie M:
Molecular basis of ocular abnormalities associatedwith proximal renal tubular acidosis. The Journal of
Clinical Investigation.108:107-115,2001.
9. Chiba Y, Ogita T, Ando K, Fujita T: PPARγ ligands
inhibit TNF-α-induced LOX-1 expression in cultured endothelial cells. Biochem Biophys Res
Commun 286: 541-546, 2001.
10. Nagase M, Ando K, Nagase T, Kaname S, Sawamura
T, Fujita T: Redox-sensitive regulation of LOX-1 gene
expression in vascular endothelium. Biochem
Biophys Res Commun 281: 720-725, 2001.
11. Chang H. Fujita T: PicSNP:a browsable catalog ofnonsynonymous single nucleotide polymorphisms in
the human genome. Biochem Biophys Res Commun287(1):288-91, 2001.
12. Sogabe H. Nangaku M. Ishibashi Y. Wada T. Fujita T.
Sun X. Miwa T. Madaio MP. Song WC: Increased
susceptibility of decay-accelerating factor deficient
mice to anti-glomerular basement membrane
glomerulonephritis. Journal of Immunology.
167(5):2791-7, 2001.

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13. Sekine N. Fukumoto S. Ishikawa T. Okazaki T. Fujita
T: GH inhibits interferon-gamma-induced signaltransducer and activator of transcription-1 activationand expression of the inducible isoform of nitricoxide synthase in INS-1 cells. Endocrinology.
142(9):3909-16, 2001.
14. Shao J. Miyata T. Yamada K. Hanafusa N. Wada T.
Gordon KL. Inagi R. Kurokawa K. Fujita T. Johnson
RJ. Nangaku M: Protective role of nitric oxide in amodel of thrombotic microangiopathy in rats.
Journal of the American Society of Nephrology.
12(10):2088-97, 2001.
15. Noiri E. Nakao A. Uchida K. Tsukahara H. Ohno M.
Fujita T. Brodsky S. Goligorsky MS: Oxidative andnitrosative stress in acute renal ischemia: American
Journal of Physiology - Renal Fluid & Electrolyte
Physiology. 281(5):F948-57, 2001.
16. Watanabe S. Fukumoto S. Takeuchi Y. Fujita H.
Nakano T. Fujita T: Effects of 1-year treatment withfluvastatin or pravastatin on bone. American Journalof Medicine. 110(7):584-7, 2001
17. Nagase M. Ando K. Nagase T. Kaname S. Sawamura
T. Fujita T:.Redox-sensitive regulation of lox-1 gene
expression in vascular endothelium. Biochem
Biophys Res Commun 281(3):720-5, 2001.
18. Kanda H. Kubo K. Hamasaki K. Kanda Y. Nakao A.
Kitamura T. Fujita T. Yamamoto K. Mimura T:
Influence of various hemodialysis membranes on theplasma (1-->3)-beta-D-glucan level. Kidney
International. 60(1):319-23, 2001
19. Yamada K. Hori Y. Hanafusa N. Okuda T. Nagano N.
Choi-Miura NH. Couser WG. Miyata T. Kurokawa K.
Fujita T: Nangaku M. Clusterin is up-regulated inglomerular mesangial cells in complement-mediatedinjury. Kidney International. 59(1):137-46, 2001.
20. Suzuki J. Ohnsihi H. Shibata H. Wada A. Hirayama
T. Iiri T. Ueda N. Kanamaru C. Tsuchida T. Mashima
H. Yasuda H. Fujita T: Dynamin is involved inhuman epithelial cell vacuolation caused by the
Helicobacter pylori-produced cytotoxin VacA.
Journal of Clinical Investigation. 107(3):363-70, 2001.
21. Kanamaru C. Yasuda H. Takeda M. Ueda N. Suzuki J.
Tsuchida T. Mashima H. Ohnishi H. Fujita T: Smad7
is induced by norepinephrine and protects rat hepatocytes from activin A-induced growth inhibition.
Journal of Biological Chemistry. 276(49):45636-41,
2001
22. Chang H. Fujita T: A numerical model of acid-basetransport in rat distal tubule. American Journal of
Physiology - Renal Fluid & Electrolyte Physiology.
281(2):F222-43, 2001.
23. Ishikawa T, Sekine N, Hata K, Igarashi T, Fujita T:
Prostagladin A1 enhances c-fos expression and activating protein-1 activity. Molecular and Cellular
Endocrinology 164:77-85,2000.
24. Sekine N, Ishikawa T, Okazaki T, Hayashi M,
Wollheim C.B., Fujita T: Synergitic activation of NFκB and inducible isoform of nitric oxide synthaseinduction by interferon-γ and tumor necrosis factor-α in INS-1 cells. J.Cell Physiol.184:46-57,2000.

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25. Chikatsu N, Takeuchi Y, Tamura Y, Fukumoto S,
Yano K, Tsuda E, Ogata E, Fujita T: Interactions
between cancer and bone marrow cells induce osteoclast differentiation factor expression and osteoclastlike cell formation in vitro. Biochem Biophys Res
Commun 267:632-637,2000.
26. Noiri E, Yokomizo T, Nakao A, Izumi T, Fujita T,
Kimura S, Shimizu T: An in vivo approach showing
the chemotantic activity of leukotrene B4 in acuterenal ischemic-reperfusion injury. ProcNatl Acad Sci.
97:823-828,2000.
27. Chikatsu N, Fukumoto S, Takeuchi Y, Suzawa M,
Obara T, Matsumoto T, Fujita T: Cloning and characterization of two promoters for the human calciumsensing receptor(CaSR) and changes of CaSR expression in parathyroid adenomas. J Biol Chem 275:75537557,2000.
28. Kunimi M, Seki G, Hara C, Taniguchi S, Uwatoko S,
Goto A, Kimura S, Fujita T: Dopamine inhibits renal
Na＋:HCO3- contransporter in rabbits and normotensive rats but not in spontabeously hypertensive rats.
Kidney International 57:534-543,2000.
29. Kubota N, Terauchi Y, Miki H, Tamemoto H,
Yamauchi T, Komeda K, Satoh S, Nakano R, Ishii C,
Sugiyama T, Eto K, Tsubamoto Y, Okuno A
Murakami K, Sekihara H, Hasegawa G, Naito M,
Toyoshima Y, Tanaka S, Shiota K, Kitamura T, Fujita
T, Ezaki O, Aizawa S, Nagai R, Tobe K, Kimura S,
and Kadowaki T.: PPARγ mediates high-fat dietinduced adipocyte hypertrophy and insulin resistance. Mol Cell. 4:597-609,1999.
30. Hori Y, Yamada K, Hanafusa N, Okuda T, Okada N,
Miyata T, Couser W, Kurokawa K, Fujita T, Nangaku
M: Crry, a complement regulatory protein, modulates
renal interstitial disease induced by proteinuria.
Kidney International 56:2096-2106,1999.
31. Chang H, Fujita T: A kinetic model of the thiazidesensitive Na-Cl cotransporter. Am J Physiol
276:F952-F959,1999.
32. Chang H, Fujita T: A numerical model of the renal
distal tubule.Am J Physiol 276:F931-F951,1999.
33. Hayashi M, Shimosawa T, Fujita T: Hyperglycemia
increases vascular adrenomedullin expression.
Biochem Biophys Res Commun 258: 453-456,1999.
34. Yasufuku-Takano J, Takano K, Takei T, Fukumoto S,
Teramoto A, Takakura K, Yamashita N, Fujita T :Heterozygous gsp mutation renders ion channnels ofhuman somatotroph adenoma cells unresponsive to
growth hormone-releasing hormone. Endocrinology
140: 2018-2026, 1999.
35. Suzuki Y, Itakura M, Kashiwagi M, Nakamura N,
Matsuki T, Sakuta H, Naiyo N, Takano K, Fujita T,
Hirose S: Identification by differential display of a
hypertonicity-inducible inward rectifier potassiumchannnel highly expressed in chloride cells. J
Biological Chem 274: 11376-11382,1999.
36. Suzawa M, Takeuchi Y, Fukumoto S, Kato S, Ueno N,
Miyazono K, Matsumoto T, Fujita T: Extracellularmatrix-associated bone morphogenetic proteins areessential for differentiation of murine osteoblastic
cells in vitro. Endocrinology 140: 2125-2133, 1999.
37. Takano K, Fujita T : Proadrenomedullin N-terminal
20 peptide hyperpolarizes the membrane by activating an inwardly rectifying K+ cyrrent in differentiated
PC12 cells.Circ Res. 84:445-450,1999.
38. Nagase M, Kato A, Ono T, Suzuki Y, Hirose S, Fujita
T : Retrotransposons transcribed preferentially inproximal tubules of salt-hypertensive rats. Kidney
International 55: 995-1004, 1999.
39. Okazaki R, Chikatsu N, Nakatsu M, Kakeuchi Y,
Ajima M, Miki J, Fujita T, Arai M, Totsuka Y,
Tanaka K, Fukumoto S :A novel activating mutationin calcium-sensing receptor gene associated with afamily of autosomal dominant hypocalcemia. J Clin
Endocrinol Metab 84: 363-366,1999.
40. Iiri T, Bell S.M., Baranski T.J., Fujita T, Bourne H.R.
:A Gsα mutant designed to inhibit receptor signaling
through Gs. Proc Natl Acad Sci. 96: 499-504, 1999.
41. Iiri T, Bell S.M., Baranski T.J., Fujita T, Bourne H.R.:A Gsα mutant designed to inhibit receptor signaling
through Gs. Proc Natl Acad Sci. 96: 499-504, 1999.
42. Isshiki M, Ando J, Korenaga R, Kogo H, Fujimoto T,
Fujita T, Kamiya A: Endothelial Ca2+ waves preferentially originate at specific loci in caveolin-rich celledges. Proc Natl Acad Sci 95:5009-5014,1998.
43. Nagase M, Abe J, Takahashi K, Ando J, Hirose S,
Fujita T : Genomic organization and regulation of
expression of the lectin-like oxidized low-density
lipoprotein receptor(LOX-1) gene. J Biological Chem
273:33702-33707,1998.
44. Hayashi M, Shimosawa T, Isaka M, Yamada S, Fujita
R, Fujita T: Plasma adrenomedullin in diabetes.
Lancet 350: 1449-1450, 1997.
45. Nakaoka T, Gonda K, Ogita T, Otawara-Hamamoto
Y, Okabe F, Kira Y, Harii K, Miyazono K, Takuwa Y,
Fujita T: Inhibition of Rat Vascular Smooth Muscle
Proliferation in Vitro and in Vivo by BoneMorphogenetic Protein-2. J Clin Invest 100: 28242832,1997.
46. Takano K, Yasufuku-Takano J, Teramoto A, Fujita T:
Corticotropin-releasing hormone excites adrenocorticotropin-secreting human pituitary adenoma cells by
activating a nonselective cation current. J Clin Invest
98:2033-2041,1996.
47. Takano K, Yamashita N, Fujita T: Proadrenomedullin
NH2-terminal 20 peptide inhibits the voltage-gated
Ca2+ channel current through a pertussis toxin-sensitive G protein in rat pheochromocytoma-derived PC
12 cells. J Clin Invest 98:14-17,1996.
48. Chung U, Tanaka Y, Fujita T: Association ofInterleukin 6 and hypoaldosteronism in patientswith cancer. New Engl J Med 334:473,1996.
49. Shimosawa T, Ito Y, Ando K, Kitamura K, Kangawa
K, Fujita T: Proadrenomedullin N-terminal 20 peptide, a new product of the adrenomedullin gene,
inhibits norepinephrine overflow from nerve endings. J Clin Invest 96:1672-1676,1995.
50. Kitagawa M, Chang H, Fujita T: Hyperkalemia due tonafamostat mesilate. New Engl J Med 332:687,1995

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Department ofAllergy and Rheumatology
Outline and Research Objectives
Our department was established in 1998 according to the reorganization of the Internal
Medicine. Doctors from previous Department of Medicine and Physical therapy and theThird Department of Internal Medicine have joined our department. Our main clinical field
are rheumatology and allergy. Therefore, our research fields also cover the areas including
basic immunology, clinical immunology, rheumatology and allergy. Especially, an area called
applied immunology such as developing new therapeutic methods has ready became important and some projects in these directions have thus been carried out in our department.
Our Reumatology and Immunology field comprises several projects including T cell receptor, autoantibody, oral tolerance, cell signaling and genetic analysis. The allergy field comprises projects of airway inflammation and IgE response.

Faculties and Students
Professor and Chair

Kazuhiko Yamamoto, M.D.
(Rheumatology, Allergology)1997
Lecturer Hirokazu Okudaira, M.D. (Allergology)
Fujio Takeuchi, M.D. (Rheumatology)
Yosikata Misaki, M.D. (Rheumatology)
Zen-ichiro Honda, M.D.(Rheumatology)
Associate .................................8
Postdoctoral Fellow ................7
Graduate student ..................16
Research student.....................2
Secretary .................................4

Past Research and Major Accomplishments
Rheumatology and Immunology
1. T Cell Receptor (TCR) Project
We have established a method for analyzing T-cellclonality using RT-PCR of TCR messages and subsequent electrophoretic separation of the PCR productbased on their single strand conformation polymorphism (SSCP). Using this method an analysis of PBLs
obtained from a healthy individual revealed the heterogeneity of the CDR3 region of the TCR beta messages. On the other hand, clonally accumulated T cells
could be identified as distinct bands on the background heterogeneous electrophoretic patterns. With
this method, we reported the importance of antigenspecific T cell clones in the pathogeneses of RA, SLE,
other autoimmune diseases.
Having observed the importance of antigen-specific T cells in autoimmune disorders, we are now trying

176

to establish methods to generate regulatory T cells
engineered by transfer of genes, the information ofwhich could be obtained from an individual patient
sample. At first, we generated an efficient alpha/beta
TCR gene transfer system using two independent
monocistronic retrovirus vectors. This system enabled
us to express the clonotypic TCR in about 25-40% of
the CD4+ T cells from murine splenocytes. The transduced cells showed an antigen-specific proliferation invitro as well as an antigen-specific delayed-type
hypersensitivity in vivo. Furthermore, regulatory T
cells engineered by the transfer of collagen-specific
TCR and IL-10 genes suppressed collagen induced
arthritis in mice. In addition, nucloesome-specific T
cells generated by the similar triple gene transfer of
TCR and CTLA4Ig were applied to lupus-prone
NZB/W F1 mice. These cells suppressed the related
pathogenic autoantibody production and nephritis
without impairing the T cell-dependent humoral
immune responses. Thus, from these results, we
could obtain efficient TCR function by gene transfer.
2. Autoantibody Project
We have been investigating in the regulatory
mechanism of autoantibody production, which is thehallmark of systemic autoimmune diseases such assystemic lupus erythematosus. We generated a transgenic mice which expresses human U1snRNP-A
autoantigen. We found that splenic T cells from thistransgenic mice suppressed the immune response tohuman U1-A. In order to further examine the regulation of autoreactive T cell in a clone level, We also
generated a transgenic mice expressing ovalbumin(OVA) systemically in nucleus. In the combination
with OVA-specific T cell receptor transgenic mice, wedemonstrated that dendritic cells are able to tolerize
autoreactive T cells. Furthermore, we found that selfreactive T cells matured into CD4(+)CD25(+) regulatory T cells in the thymus.

2) Linkage analysis using a familial juvenile hyperuicemic nephropathy (FJHN) revealed the susceptibility region in chr16p12.

3. Oral Tolerance Project
We have studied the mechanism of tolerance
induction in the mucosal immune system and its
application to therapy of autoimmune diseases.
Especially, we focused on the role of DC (dendritic
cells) for oral tolerance after antigen feeding. We
examined the requirement for tolerance induction in
adoptive cell transfer system and found that not only
T cells but DCs in Peyer’s patches or spleen are important for the transfer of oral tolerance. We also fed
OVA to Balb/c mice in bronchial asthma mouse model
induced by OVA and confirmed that OVA feeding
suppressed airway hypereactivity. In this system,
depletion of DCs abrogated the transfer of tolerance.

Allergy
1. Airway Inflammation Project
We established a system to evaluate antigen specific response of the airway in the mouse under a noninvasive, unrestrained condition. Using this system,
we analyzed relationships among sensitization to anallergen, increase in airway hyperreactivity (AHR),
and development of airway inflammation. The results
demonstrated that contrary to the previous general
understandings, AHR can be induced with systemic
sensitization alone (early phase AHR).
We have also found that intratracheal administration of hepatocyte growth factor (HGF), a regenerative
growth factor for bronchial and alveolar epithelial
cells could attenuate extent of the experimental lungfibrosis. In addition, we elucidated the mechanism ofaction and found that HGF has a potent migratory
activity and enhancing activity of surface fibrinolytic
system of lung epithelial cells. Further, we reported
that anti-fibrotic cytokine IFN-γ potentiates the biological activity of HGF by upregulating the expressionof c-Met/HGF receptor on alveolar epithelial cells.

4. Intracellular Signaling Project
Receptors Fc portion of Immunoglobulines play
central roles in the initial step of immune regulation.
For the cells to be activated, the receptors should becross-linked by multivalent ligands. We have engaged
in solving the mechanisms, in which physical receptor
cross-linking is converted into biochemical signaling.
It was assumed that receptor engagement is sensed
and converted by Src family kinases (SFKs) to cellinner events. However, the compensatory function ofeight SFKs hampered clear demonstration of thenotion. We introduced novel strategies to circumventthe difficulty, and established their requirement andspecificity in Fc, and c-Kit and Integrin receptors.
Notably, SFKs responsible for Fc receptor signaling are
all modified with palmitic acid, and they reside in amembrane domain referred to as ‘lipid rafts’.
On the other hand, we found that a 105 kDa
moledule was tyrosine phosphorylated through TCR
ligation in human T cells. It was proven to be p105CasL,
a member of the p130Cas-related docking protein family, which subsequently binded to the Src homology 2
domain of c-Crk. We showed that it was also tyrosine
phosphrylated though integrin stimulation in T cells,
and its expression was predominant in lymphocytes.
Next, we reported that Fyn and Lck tyrosine kinases
regulated tyrosine phosphorylation of p105CasL in TCRmediated signaling in human T cells and splenocytesof lpr mice, SLE model mice.
5. Genetics Project
We have studied genetic background and pathophysiology of rheumatic diseases, and mechanisms
and effect of aging were studied. Examples are1)Genetic analysis of HLA-DRB1 shared epitope in
RA family cases showed the participation of the epitope was neither unique nor indispensable.

2. IgE Response Project
Surface expression of high affinity IgE receptor(FceRI) on mast cells is essential for these cells toexert IgE-dependent effector functions. We have analyzed how and to what extent mast cell surface FceRI
expression is regulated, and whether regulation of
their surface FceRI levels is functionally important.
Through the series of our study, we found that the ligand of FceRI, IgE, which is known to stabilize FceRI,
can greatly upregulate the surface FceRI levels onmast cells. A Th2 cytokine, IL-4, is also involved in
FceRI upregulation in mast cells. Mast cells withupregulated levels of surface FceRI in the presence of
IgE and/or IL-4 demonstrate enhancement in IgEdependent mediator release and cytokine production.
On the other hand, glucocorticoid, decreases the surface FceRI levels of mouse mast cells either in thepresence or absence of IgE, via posttranscriptional
mechanism. These results indicate that mast cell surface FceRI levels are regulated via multiple mechanisms by various factors including IgE, cytokines andglucocorticoids, resulting in functional alteration ofmast cells. IgE- and FceRI-related positive feedback
mechanism may account for the pathogenesis of IgEmediated allergy.

177Current Research
Rheumatology and Immunology
1. TCR Project
We are now trying to establish an efficient method
to reconstitute TCR function using information available from a small sample of a patient. In order toaccomplish these projects, two important technological breakthroughs are necessary. They are cloning
techniques of full length cDNA encoding alpha andbeta TCR from a single cells and efficient vectors todeliver these genes into usual lymphocytes. We are
now trying to develop several possible technicalimprovements.
2. Autoantibody Project
We have identified several unique molecules to
CD4(+)CD25(+) regulatory T cells including transcription factors and proliferation-associated molecules,
which indicate that the subset is a unique and distinct
population.
3. Oral Tolerance Project
We are studying the mechanisms of generation ofregulatory T cells which mediate mucosal tolerance.
We are visualizing the interaction between T cells and
DCs which captured antigen in Peyer’s patch and characterizing DCs which are involved in generation ofregulatory T cells. We are also investigating a role of
TGF-β for generation of regulatory cells in mucosal
tolerance and for class switching of B cells to IgA
secreting plasma cells.
4. Intracellular Signaling Project
We are searching for the molecules responsible for
Fc receptor-mediated lipid raft assembly through protein chemistry approaches. We have also cloned two
novel molecules interacting with a SFK, Lyn. One isan ubiquitin ligase with RING domain, presumably
promoting SFK turnover. Another is a huge adaptor
protein possessing putative Grb2 and PI3K bindingsites and GAP domain.
We are also investigating whether IGF-I could prevent glomerulosclerosis, because glomerulosclerosis
was not found in IGF-I overexpressing mice, althoughglomerulohypertrophy became obvious. To investigate
this, we are evaluating the effects of adding IGF-I after
adhesion stimulation on some signaling molecules in
rat mesangial cells. Concurrently, we are preparing an
animal study, using 5/6 nephrectomy as glomeruloscrelosis model rat, to know whether IGF-I infusion
prevents glomerulosclerosis.
5. Genetics ProjectSome candidate gene analysis are now in progress

178

Allergy
1. Airway Inflammation Project
We have recently found that passive cell transferof spleen cells obtained from sensitized mouse into
naive mouse provokes an increase in the AHR. This
indicates that mechanisms to induce AHR can bestudied. We are now analyzing which kind of cell in
the lung, such as antigen presenting cells, mast cells,
and lymphocytes, would contribute to the development of early phase AHR by negative and positive
selection of spleen cells.
2. IgE Response Project
We are interested whether IgE crosslinking stimulation of mast cells and basophils can result in cellularchanges other than activation. Currently we are
exploring the phenomenon of FceRI-mediated desensitization. At present we think that cellular desensitization and stimulation may be separate but closely
related to each other.

Future Prospects
Rheumatology and Immunology
1. TCR Project
Our final goal is to establish an antigen- specificimmunotherapy in terms of TCR gene transfer using
information from a small sample of an individual
patient. Once the basic system is established, we
could apply this to several different immunologicaldisorders. Applicable disorders include rheumatoid
arthritis, systemic lupus erythematosus and otherconnective tissue diseases, cancer patients, transplantation, severe infections such as AIDS and severe
forms of allergy. We believe that this system would
become one of fundamental methods in clinicalimmunology.
2. Autoantibody Project
We will clarify the generative mechanism of regulatory T cells generation and will innovate the methods to control the regulatory T cells.
3. Oral tolerance Project
Our purpose is to treat autoimmune diseases using
the mechanism of tolerance induction by mucosal
immune system.
4. Intracellular Signaling Project
Identification of the ‘raft target’ is unequivocallyimportant for basic science and for therapeutic intervention of inflammatory diseases.
The pathological examination of end-stage renal
disease shows similar glomerulosclerotic patterns,although there are many causes of renal diseases.
Therefore, we believe that preventing glomerulosclerosis could stop the progression to end-stage renal disease.
5. Genetics Project
Genes wide screen of several diseases would be
expected.

Allergy
1. Airway Inflammation Project
We are expecting to establish a new strategy forpreventing the development of AHR before onset of
airway inflammation.
2. IgE Response Project
The aim of our study is to understand the pathogenesis and establish the new therapeutic strategy of
IgE-mediated allergic diseases, such as bronchial asthma and allergic rhinitis, from which more people aresuffering. Because IgE-mediated allergy seems much
more complicated than previously thought, our future
progress in this research area will be of substantial
importance.

Research Grants
1. Research on Specific Diseases from the Ministry of
Health and Welfare of Japan 1999-2001.
2. Research on Allergic disease and Immunology from
the Ministry of Health, Labour and Welfare 20022004
3. Research on Health Sciences focusing on Drug
Innovation from the Japan Health Sciences
Foundation 2000-2002
4. Research on Eye and Ear Sciences, Immunology,
Allergy and Organ Transplantation from the
Ministry of Health and Welfare of Japan 1998-2000.
5. Research Grant from the Ministry of Education,
Science, Sports and Culture of Japan(Kibann
KenkyuA) 1999-2001

Select Publications
1. Kawahata K, Misaki Y, Yamaguchi M, Tsunekawa S,
Setoguchi K, Miyazaki J and Yamamoto K. Peripheral
tolerance to a nuclear autoantigen:dendritic cells
expressing a nuclear autoantigen lead to persistentanergic state of CD4 autoreactive T cells after proliferation. J Immunol.168:1103-1112, 2002.
2. Nagase H, Miyamasu M, Yamaguchi M, Imanishi M,
Tsuno NH, Matsushima K, Yamamoto K, Morita Y
and Hirai K. Cytokine-mediated regulation of CXCR4
expression in human neutrophils. J Leukocyte
Biol.71:711-717, 2002.

3. Yoshimura C, Yamaguchi M, Iikura M, Izumi S, Kudo
K, Nagase H, Ishii A, Wall AF, Ra C, Iwata T, Igarashi
T, Yamamoto K and Hirai K. Activation makers ofhuman basophils:CD69 expression is strongly andpreferentially induced by IL-3. J Allergy Clin
Immunol.109:817-823, 2002.
4. Kawahata K, Misaki Y, Yamauchi M, Tsunekawa S,
Setoguchi K, Miyazaki J and Yamamoto K.
Generation of CD4+CD25+ regulatory T cells fromautoreactive T cells simultaneously with their negative selection in the thymus and from non-autoreactive T cells by endogenous T cell receptor expression. J Immunol. 68:4399-405, 2002.
5. Kono H, Suzuki T, Yamamoto K, Okada M, Yamamoto
T and Honda Z. Spatial raft coalescence represents
an initial step in Fc gamma R signaling. J
Immunol.169:193-203, 2002.
6. To Y, Dohi M, Matsumoto K, Tanaka R, Sato A,
Nakagome K, Nakamura T and Yamamoto K. A two
way interaction between hepatocyte growth factor
and interleukin-6 on tissue invasion of lung cancer
cell line. Am J Respir Cell Mol Biol.27:220-226, 2002.
7. Takami K, Takuwa N, Okazaki H, Kobayashi M,
Ohtoshi T, Kwasaki S, Dohi M, Yamamoto K,
Nakamura T, Tanaka M, Nakahara Y and Takizawa
H. Interferon-gamma inhibits hepatocyte growth factor-stimulated cell proliferation of human bronchial
epithelial cells.Upregulation of p27 (kip1) cyclindependent kinase inhibitor. Am J Respir Cell Mol
Biol.26:231-238, 2002.
8. Misaki Y, Ezaki I, Ariga T, Kawamura N, Sakiyama Y
and Yamamoto K. Gene-transferred oligoclonal T
cells predominantly persist in peripheral blood froman adenosine deaminase- deficinent patient during
gene therapy. Mol Ther.3:24-27, 2001.
9. Yamaguchi M, Hirai K, Komiya A, Miyamasu M,
FukumotoY, Teshima R, Ohta K, Morita Y, Galli SJ,
Ra C and Yamamoto K. Regulation of mouse mastcell surface F e RI expression by dexamethasone. Int
Immunol.13:843-851, 2001.
10. Yoshimura C, Miyamasu M, Nagase H, Iikura M,
Yamaguchi M, Kawanamai O, Morita Y, Iwata T,
Yamamoto K and Hirai K. Glucocorticoids induce
basophil apoptosis. J Allergy Clin Immunol.108:201220, 2001.
11. Yamada R, Tanaka T, Unoki K, Nagai T, Sawada T,
Ohnishi Y, Tsunoda T, Yukioka M, Maeda A, Suzuki
K, Tateishi H, Ochi T, Nakamura Y and Yamamoto
K. Association between a single-nucleotide polymorphism in the promoter of the human interleukin-3
gene and rheumatoid arthritis in Japanese patients,and maximum-likelihood estimation of combinatorial effect that two genetic loci have on susceptibility
to the disease. Am J Hum Genet.68:674-685, 2001.
12. Kawasaki S, Takizawa H, Yoneyama H, Nakayama T,
Fujisawa R, Izumizaki M, Imai T, Yoshie O,
Hommma I, Yamamoto K and Matsushima K.
Intervention of thymus and activation-regulated
chemokine attenuates the development of allergic

179airway inflammation and hyperrespnsiveness inmice. J Immunol.166:2055-2062, 2001.
13. Setoguchi K, Misaki Y, Terauchi Y, Yamauchi T,
Kawahata K, Kadowaki K and Yamamoto K.
Peroxisome proliferator-activated receptor-γ haploinsufficiency enhances B cell proliferative responses and exacerbates experimentally induced arthritis.
J Clin Invest.108:1667-1675, 2001.
14. To Y, Dohi M, Tanaka R., Sato A, Nakagone K, andYamamoto K. Early IL-4-dependent response can
induce airway hyperreactivity in amouse model ofasthma. Lab Invest.81:1385-96, 2001.
15. Kanda H, Kubo K, Hamasaki K, Kanda Y, Nakao A,
Kitamura T, Fujita T, Yamamoto K, Mimura T.
Influence of various hemodialysis membrane on theplasma (1→3)-β-D-glucan level. Kidney Int.60:31923, 2001.
16. Nagase H, Miyamasu M, Yamaguchi M, Fujisawa T,
Ohta K, Yamamoto K, Morita Y and Hirai K.
Expression of CXCR4 in eosinophils:Functional
analyses and cytokine- mediated regulation. J
Immunol.164:5935-43, 2000.
17. Sasaki M, Nakamura S, Ohyama Y, Shinohara M,
Ezaki I, Hara H, Kadena T, Kishihara K, Yamamoto
K, Nomoto K and Shirasuna K. Accumulation of common Tcell clonotypes in the salivary glands ofpatients with human T lymphotropic virus typeIassociated and idiopathic Sjögren’s syndrome. J
Immunol.164:2823-2831, 2000.
18. Fujio K, Nosaka T, Kojima T, Kawashima T, Yahata
T, Copeland NG., Gilbert DJ, Jenkins NA, Yamamoto
K, Nishimura T and Kitamura T. Molecular cloning of
a novel type 1 cytokine receptor similar to the common gamma chain. Blood.95:2204-2210, 2000.
19. Sawabe T, Shiokawa S, Sugisaki K, Tsuda T and
Yamamoto K. Accumulation of common clonal T
cells in multiple lesions of sarcoidosis. Mol
Medicine.6:793-802, 2000.
20. Sekiya T, Miyamasu M, Imanishi M, Yamada H,
Nakajima T, Yamaguchi M, Fujisawa T, Pawankar R,
Sano Y, Ohta K, Ishii A, Morita Y and Yamamoto K.
Matsushi K. Yoshie O. and Hirai K. Inducible expression of a Th2-type CC chemokine thymus- and activation-regulated chemokine (TARC) by humanbronchial epithelial cells. J Immunol.165:2205-2213,
2000.
21. Nagase H, Yamaguchi M, Jibiki S, Yamada H, Ohta K,
Kawasaki H, Yoshie O, Yamamoto K, Morita Y andHirai K. A rapid and simple photometric assay forquantification of eosinophil chemotaxis. Int Arch
Allergy Immunol.122:10-14, 2000.
22. Fujio K, Misaki Y, Setoguchi K, Morita S, Kawahata
K, Kato I, Nosaka T, Yamamoto K and Kitamura T.
Functional reconstitution of classII MHC-restricted
Tcell immunity mediated by retroviral transfer of thea b TCR complex. J Immunol.165:528- 532, 2000.
23. Setoguchi K, Misaki Y, Araki Y, Fujio K, Kawahata K,
Kitamura T and Yamamoto K. Antigen-specific T
cells transduced with IL-10 ameliorate experimentally induced arthritis without impairing the systemic

180

immune response to the antigen. J Immunol.
165:5980-5986, 2000.
24. Nagase H, Miyamasu M, Yamaguchi T, Kawasaki H,
Ohta K, Yamamoto K, Morita Y and Hirai K.
Glucocorticoids preferentially upregulate functional
CXCR4 expression in eosinophils. J Allergy Clin
Immunol.106:1132-1139, 2000.
25. Suzuki T, Kono H, Hirose N, Okada M, Yamamoto T,
Yamamoto K and Honda Z. Differential involvementof Src family kinases in Fc gamma receptor-mediatedphagocytosis. J Immunol.165:473-482, 2000.
26. Honda Z, Suzuki T, Kono H, Okada M, Yamamoto T,
Ra C, Morita Y and Yamamoto K. Sequential requirements of the N-terminal palmitoylation site and SH2
domain of Src family kinases in the initiation andprogression of Fcepsilon RI signaling. Mol Cell
Biol.20:1759-1771, 2000.
27. Furukawa H, Yabe T, Watanabe K, Miyamoto R, Miki
A, Akaza T, Tadokoro K, Tohma S, Inoue T,
Yamamoto K and Juji T. Tolerance of NK and LAK
activity for HLA class I-deficient targets in a TAPdeficient patient. (bare lymphocyte syndrome type I)
Hum Immunol.60:32-40, 1999.
28. Matsui T, Kurokawa M, Kobata T, Oki S, Azuma M,
Tohma S, Inoue T, Yamamoto K, Nishioka K and
Kato T. Autoantibodies to Tcell co-stimulatory molecules in systemic autoimmune diseases. J
Immunol.162:4328-4335, 1999.
29. Furukawa H, Murata S, Yabe T, Shimbara N, Keicho
N, Kashiwase K, Watanabe K, Ishikawa Y, Akaza T,
Tadokoro K, Tohma S, Inoue T, Tokunaga K,
Yamamoto K, Tanaka K and Juji T. Splice acceptor
site mutation of transporter associated with antigen
processig-1 gene in human bare lymphocyte syndrome. J Clin Invest.103:755-758,1999.
30. Nagahori T, Dohi M, Matsumoto K, Saitoh K, Honda
ZI, Nakamura T and Yamamoto K. Interferon-gamma
upregulates the c-Met/hepatocyte growth factor
receptor expression in alveolar epithelial cells. Am J
Resp Cell Mol Biol.21:490-497, 1999.
31. Kawahata K, Misaki Y, Komagata Y, Setoguchi K,
Tsunekawa S, Yoshikawa Y, Miyazaki J andYamamoto K. Altered expression level of a systemic
nuclear autoantigen determines the fate of immune
responce to self. J Immunol.162:6482-6491, 1999.
32. Furukawa H, Yabe T, Akaza T, Tadokoro K, Tohma
S, Inoue T, Tokunaga K, Yamamoto K, Geraghty DE
and Juji T. Cell surface expression of HLA-E molecules on PBMC from a TAP1-deficient patient. Tissue
Antigens.53:292-295, 1999.
33. Takizawa H, Ohtoshi T, Kawasaki S, Kohyama T,
Desaki M, Kasama T, Kobayashi K, Nakahara K,
Yamamoto K, Matsushima K and Kudoh S. Diesel
exhaust particles induce NF-κB activation in humanbronchial epithelial cells in vitro:Importance incytokine transcription. J Immunol.162:47054711,1999.
34. Miyamasu M, Yamaguchi M, Nakajima T, Misaki Y,
Morita Y, Matsushima K, Yamamoto K and Hirai K.
Th1-derived cytokine IFN-γ is a potent inhibitor ofeotaxin sythesis in vitro. Int Immunol.11:1001-1004,
1999.
35. Iikura M, Yamaguchi M, Miyamasu M, Morita Y,
Iwase T, Moro I, Yamamoto K and Hirai K. Secretory
IgA-mediated basophil activation. Biochem BiophRes Co.264:575- 579, 1999.
36. Miyamasu M, Nakajima T, Misaki Y, Izumi S, Tsuno
N, Kasahara T, Yamamoto K, Morita Y and Hirai K.
Dermal fibroblasts represent a potent major sourceof human eotaxin:in vitro production and cytokinemediated regulation. Cytokine.11:751-758, 1999.
37. Murata S, Kawahara H, Tohma S, Yamamoto K,
Kasahara M, Nabeshima Y, Tanaka K and Chiba T.
Growth retardation in mice lacking the proteasome
activator PA28g. J Biol Chem.274:38211-38215, 1999.
38. Kanda H, Mimura T, Hamasaki K, Yamamoto K,
Yazaki Y, Hirai H and Nojima Y. Fyn and Lck tyroine
kinase regulate tysine phoshorylation of p105CasL, amember of the p130Cas docking protein family,in Tcell receptor-mediated signalling. Immunol. 97:56-61,
1999.
39. Kubo K, Miyagawa K, Yamamoto R, Hamasaki K,
Kanda K, Fujita T, Yamamoto K, and Mimura T.
Detection of WT1 mRNA in urine from Patients withkidney diseases. Eur J Clin Invest.29:824-826, 1999.
40. Dohi M, Tsukamoto S, Nagahori T, Shinagawa K,
Saitoh K, Tanaka Y, Kobayashi S, Tanaka R, To andYamamoto K. Noninvasive system for evaluating theallergen- pecific airway response in a murine modelof asthma. Lab Invest.79:1559-15, 1999.
41. Furukawa H, Yabe T, Watanabe K, Miyamoto R,
Akaza T, Tadokoro K, Tohma S, Inoue T, Yamamoto
K and Juji T. An alternatively spliced form of thehuman CD94 gene. mmunogenetics.48:87-88, 1998.
42. Nakajima T, Yamada H, Iikura M, Miyamasu M,
Izumi S, Shida H, Ohta K, Imai T, Osamu Y,
Mochizuki M, Schroder J, Morita Y, Yamamoto K
and Hirai K. Intracellular localization and release of
eotaxin from normal eosinophils. FEBS Lett.434:226230, 1998.
43. Miyazawa K, Mori A, Yamamoto K and Okudaira H.
Transcriptional roles of C/EBP-β, NF-κB,and CBF/minterleukin-Iβ-induced interleukin-6 synthesis byhuman rheumatoid fibroblast-line synoviocytes. J
Biol Chem.273:7620-7627, 1998.
44. Miyazawa K, Mori A, Yamamoto K and Okudaira H.
Constitutive transcription of the human IL-6 gene byrheumatoid synoviocytes spontaneous activation of
NF-κB and CBFI. Am J Pathol.152:793-803, 1998.
45. Shiokawa S, Nishikawa J, Ohshima K, Uike N and
YamamotoK. Establishment of a novel B cell clonality analysis using single-strand conformation polymorphism of immunoglobulin light chain messenger
signals. Am J Pathol.153:1393-1400, 1998.
46. Suzuki T, Shoji S, Yamamoto K, Nada S, Okada M,
Yamamoto T and Honda Z. Essential roles of Lyn infibronectin-mediated filamentous actin assembly andcell motility in mast cells. J Immunol.161:3694-3701,
1998

47. Iikura M, Yamaguchi M, Fujisawa T, Miyamasu M,
Takaishi T, Morita Y, Iwase T, Moro I, Yamamoto K
and Hirai K. Secretory IgA induces degranulationfrom IL-3- primed basophils. J Immunol.161:15101515, 1998.
48. Uozumi N, Kume K, Nagase T, Nakatani N, Ishii S,
Tashiro F, Komagata Y, Maki K, Ikuta K, Ouchi Y,
Miyazaki J, and Shimizu T. Role of cytosolic phospholipase A2 in allergic response and parturition.
Nature.390:618-622, 1997.
49. Honda Z, Suzuki T, Hirose N, Aihara M, Shimizu T,
Nada S, Okada M, Ra C, Morita Y and Ito K. Roles of
C-terminal Src kinase in the initiation and the termination of the high affinity IgE receptor-mediated signaling. J Biol Chem.272:25753-25760, 1997

181

Department of
Psychosomatic Medicine
Outline and Research Objectives
Department of Psychosomatic Medicine was founded in 1972 at the Branch Hospital of
the University of Tokyo.
Our department started as a clinical section for seeing ‘psychosomatic diseases’ that arephysical diseases whose onset and courses are greatly affected by psychosocial, stress-related factors in the field of internal medicine. However, we have come to cover many patientsof eating disorders, panic disorders, and mild depression that are not strictly psychosomatic
diseases but share common features of the presence of many physical symptoms and theinfluence of psychosocial factors.
Based on these historical circumstances, our department has mainly focused on the
problems of eating disorders and has also studied panic disorder and mild depression.
However, we have also actively studied lifestyle-related and adult diseases such as hypertension (HT), diabetes mellitus (DM), Basedow’s disease, and cancers in order to meet thesocial needs expecting us to see stress-related diseases in the field of internal medicine,investigating what kinds of psychosocial factors are related to their onset and courses andwhat kinds of psychobehavioral interventions are effective on these diseases. Moreover, wehave performed basic and interventional studies of various relaxation procedures that could
cancel hazardous effects of stress on our health.
Because psychosomatic medicine on which we rely is rather a new field of medical science born about half a century ago and thus special research methodologies have not been
established yet, we should have developed a new method for performing new research.
Moreover, the pathological mechanisms of our subjects such as psychosomatic diseases andeating disorders are so complicated by many factors including physical, psychological andbehavioral ones, and it has been very difficult for us to empirically uncover their roles one
by one.
However, the conditions surrounding our field of research have dramatically changed inrecent years, because many new kinds of methods have been developed that can be applied
to psychosomatic field including brain functional imaging procedures, ecological momentary
assessment (EMA) which means successive monitoring of biopsychobehavioral time series
data in daily life, and the discovery and development of basic research of novel eating-related substances. Furthermore, the increase of various psychosocial stress and the acknowledgment of its hazardous effects on health in modern world heighten the social needs forthe evidence-based interventional procedures of stress-related diseases far stronger than
before. Therefore, our department must greatly contribute to our society in the near future.

Faculties and Students
Professor and Chair

Tomifusa Kuboki, MD,PhD, Since
April 1996
Associate Professors Hiroaki Kumano, MD,PhD
Associate .................................2
Graduate student ....................7
Research student.....................1
Secretary .................................3

Past Research and Major Accomplishments
We have mainly performed clinical research ofpatients consulting our department and behavioral

182

scientific research of life-related and adult diseases
consulting the department of internal medicine andother physical departments. Furthermore, as a basic
research, we have investigated one of the main subjects of psychosomatic medicine that is mind/bodycorrelation meaning interaction between biological,
psychological, and behavioral aspects of human being
through the study of stress and relaxation. The main
fields of our research are as follows and related papers
are listed in parentheses the number of which corresponds to the list of Selected Publicatios.
1) Clinical, behavioral scientific, and epidemiologicalstudiesoneatingdisorders(#1,2,3,4,5,10,11,14,16,23,25,30)

2) Behavioral scientific, psychophysiological, andinterventional studies on lifestyle-related andadult diseases including DM, HT, Basedow’s disease,andcancers(#9,13,17,18,21,22,24,26,28,29,31,33,34)
3) Studies on other clinical subjects of our department
including panic disorder and depression(#6,12,20,27,32,36,37)
4) Studies on the development of questionnaires concerning mental health including the TEG, the TSS,
the POMS, the LHQ, the SIRI33, and the SSA(#7,28,31,33,35)
5) Psychophysiological and interventional studies on
various relaxation procedures such as autogenic
training, biofeedback, photic feedback, and Qigong (#8,15,18,19,26,29,34)
Concerning eating disorders, we did the first country-wide epidemiological study and made the mannual of cognitive bahavioral therapy and family guidancefor carrying out controlled treatment study collaborating on the projects of Ministry of Health and Welfare.
Furthermore, we did many clinical studies for clarifying various features of these patients.
Concerning lifestyle-related and adult diseases, wehave introduced advanced methodologies of behaviralsciences and data analysis prodedures including various multivariate analyses. We have gathered many
data on how psychobehavioral problems such as
lifestyles, stress, and personality could affect theonset and courses of these diseases. These data areuseful for clarifying how we should change what kindof behavior for behavior modification that are regarded to be important for the treatment of these diseases.
Stress and mental health are important research
themes of psychosomatic medicine, and we investigated hazardous effects of stress by developing various psychobehavioral questionnaires. We got excellent results such as developing the Tokyo University
Egogram (TEG) that are now widely used not only in
hospitals but also in companies and schools all overJapan, and the Life Health Questinnaire (LHQ) for
assessing multiple stress-related variables based on
the data of more than 10,000 healthy people.
Furthermore, the questionnaire for examining the riskof certain personality trait on the occurrence of cancers named the Short Interpersonal Reactions
Inventory (SIRI33) was also developed, and a collaborative study with the National Cancer Center is now
in progress.
Because we speculate that the cumulative effects ofstress may precipitate or aggravate psychosomatic diseases, panic disorder, or depression, it is urgently necessary to develop therapeutic procedures soothing theeffects of stress. We have investigated various relax-

ation procedures as such stress-reducing methods. As
basic research, physical and psychological characteristics of relaxation response have been studied throughpsychophysioloical measures of autonomic nervous
system and electroencephalogram (EEG), and a questionnaire assessing psychological relaxation response
has been developed. Furthermore, we have done several interventional studies targeting DM, psyochosomatic diseases, or depression, and excellent results
have been obtained such as very positive effects of Qigong group therapy on metabolic control of type 2
DM substantiated by a randomized controlled trial(RCT).
Because psychosomatic medicine is rather a new
field of medical science, we should have developed
new methods or introduced various methods fromadjacent fields for performing new research. As mentioned above, we have achieved several positive
results in this point and have made it possible to gather and analyse relevant data, which had never beenpossible before.

Current Research
We are now advancing more basic research investigating the relationship between physical, psychological and behavioral aspects in the onset and courses ofvarious stress-related diseases using newly developed
research methodologies such as noninvasive assessment procedures of brain or autonomic nervous system function and EMA of human time-series bioinformation. Furthermore, it should be noted that targets of research have been expanded to include subjects complaining mainly fatigue who suffer fromchronic fatigue syndrome (CFS) and multiple chemicalsensitvities (MCS) and both healthy and mildly
demented geriatric poplulation.
Now we have four active research laboratories inour department, and each is pursuing various research
projects as follows making full use of unique methodologies and collaborating with many outside institutes
and researchers.
1) Behavioral Science Laboratory
Objectives: In order to clarify the disease processes and ways of treatment of lifestyle-related diseases
and psychosomatic diseases, we should deal with thefunctional aspect of subjects fucusing on behavior
meaning how they live a daily life as well as theanatomical aspect focusing on body and the introspective aspect focusing on mind. The objectives of thislaboratory are to investigate behavior as well as its
relation with mind and body of various subjects ranging from psychosomatic patients to healty persons.
Methodologies: Using questionnaires, personal
degital assistants, psychophysiological assessment

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procedures, or blood chemistry, the multiple data concerning behavior, mind and body are gathered. Then,the structure within the data is precisely extracted bymaking full use of many kinds of data analysis methods such as multivariate analyses, time-series analyses, and analysis of variance.
Projects:
DM: Cross-sectional and longitudinal studies toclarify multiple psychosocial factors affecting metabolic control and quality of life (QOL) of diabetic patients
Tension-type headache: An interventional study
by autogenic training
Cancer: A semi-prospective study of the role oftype C personality on the onset of prostate cancer; A
prospective study of personality factors affecting theonset of various cancers in healthy persons
Renal failure: A cross-sectional study on the relatonship between stress-related factors and metabolic
control and QOL of hemodialysis patients
MCS: A cross-sectional study on stress-related factors affecting their onset and courses
Depression: To determine the diagnostic criteria ofmild depression
Elderly people: Cross-sectional and longitudinalstudies to clarify the relationship between bodilyfunction, exercise ability, lifestyle, and stress-related
factors2) Bio-signal Analysis Laboratory
Objectives: Although human body produces various kinds of signals, most of them are not captured
and discarded as noise. However, as many novel signal analysis methods have been developed these days,
we can now extract valuable information from signals
previously regarded as noise and can investigate sucha field as has been difficult to be targeted by scientificinquiries. The objectives of this laboratory are tounderstand pathophysiology, mind/body correlation,
stress, or relaxation by using such novel methodologies.
Methodologies: Higher brain functions are analyzed by use of positron emission tomography (PET),
functional magnetic resonance imaging (fMRI), magnetic encephalography (MEG), and EEG. Autonomic aswell as central nervous functions are estimated byspectral and fractal analyses of heart rate variabilities.
Ecological changes of heart rate variabilities, bodily
movement, mood, symptoms, and toxic substances inthe environment are successively monitored by EMA
methods.
Projects:
PET: Measurement of cerebral blood flow or glucose metabolism before and after the treatment ofanorexia nervosa or panic disorders; Investigation on
the relationship between stress coping processes andcerebral blood flow

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EEG: Studies assessing the improvement of cerebral cortical function by some relaxation procedures
using the dipole tracing method; Assessment ofpatients with various diseases by event-related potentials (ERP)
Heart rate variabilities & EMA: A study on autonomic nervous activities and bodily movement in therefeeding phase of anorexia nervosa; Development ofautonomic nervous function database of outpatientsin our department; Comprehensive assessment ofmultiple chemical sensitivities and chronic fatigue
syndrome by EMA methods
3) Neuro-immuno-endocrinology Laboratory
Objectives: The biological basis of mind/body correlation is regarded as ‘functional association between
neuroendocrine and immune systems’, that is sought
to be clarified by clinical as well as basic methods inthis laboratory. Brain mechanism affecting abnormal
eating behavior is also a main focus to be inquired. In
addition, the search for physiological measures ofstress responses and their application to clinical field
is another challenge.
Methodologies: Methods of functional morphology and behavioral analysis are used for analyzing
genetically-modified mice such as a knockout mouse.
We take notice of 8-hydroxy-2’-deoxy guanosine(8OHdG) as an oxidated substance of DNA and measure it in human urine.
Projects:
Inflammatory cytokines: Analyses of expression
and mechanism of action in brain tissue of inflammatory cytokines such as IL-1 and IL-6 as modulators ofneuroendocrine and immune systems
Eating modulating substances: Investigation on
the effects of newly-found neuropeptides such as
orexin and ghrelin in brain on emotion and stressresponses
Novel stress measure: Investigation on the usefulness of 8OHdG as a physiological measure in humanstress-related diseases
4) Eating Disorders Laboratory
Objectives: The etiology of eating disorders suchas anorexia nervosa and bulimia nervosa wasassumed to be affected by many factors including
individual psychological, biological, familial, socio-cultural factors. The objectives of this laboratory are to
study various factors relating to the courses of eating
disorders, to develop and improve treatment procedures, and to do the basic research of pathophysiology collaborating with other laboratories.
Methodologies: Bio-psycho-behavioral clinicaldata at the initial visit and during treatment courses
are gathered; Pathophysiology of eating disorders isinvestigated by PET or EMA.

Projects:
Dropout: A study by survival analysis on premature termination of outpatient treatment
Refeeding: Assessment of energy metabolism,
autonomic nervous function, and bodily movement in
the refeeding phase of anorexia nervosa patients
Cerebral blood flow and dopaminergic metabolism:
Pet studies of anorexia nervosa patients

Future Prospects
We want to continue to advance activities ofabove-mentioned four laboratories and to focus on thefollowing points in particular.
1) Brain processes closely related to mind/body correlation in various stress-related diseases and pathogenic brain functional abnormalities will be clarified by active use of functional brain analysis
methods such as PET, fMRI, MEG, ERP, and dipole
tracing methods.
2) Ecological changes in everyday life of biological,
psychological, behavioral, and environmental
measures will be monitored and analyzed in orderto characterize neurobehavioral basis of various
stress-related diseases by developing and utilizing
relevant EMA methods.
3) Treatment programs will be developed that intervene psychobehavioral factors found to be closely
related to the processes of psychosomatic diseases
and lifestyle-related diseases by our behavioral scientific studies and include effective relaxation procedures. Then, the evidence will be shown thatthese interventions are effective in improving not
only symptoms but also QOL and life satisfaction
by carrying out RCT of these treatment programs.
4) Biological and material basis of eating disorders will
be pursued by both basic research such as one
using genetically-modified mice and clinicalresearch measuring eating-related substances inthe blood of eating disorders patients before andafter oral glucose tolerance tests.
In summary, we are going to advance toward two
directions. First, we will change over from the previous research system mainly relying on clinical andpsychobehavioral domains to the new research system that enables us to investigate neurobehavioral
and biological mechanisms actively utilizing novel
methodologies. Then, we will accumulate useful data
for the development of biological treatment by broadening our horizons from mind/body correlation to
mind/brain/body correlation and by seeking for thetrue nature of pathological processes of stress-relateddiseases.
Secondly, we will make it possible to gather
diverse data in patients’ everyday life by incorporating

EMA into behavioral scientific methodologies, andthen we will aim to clarify the reality of not only diseases themselves but also patients having such diseases. Based on these results as well as our present
knowledge, we will develop treatment programs
including interventions manipulating psychobehavioral variables and relaxation procedures and will clarify by RCT what kinds of interventions most effectively improve not only symptoms but also QOL andlife satisfaction.

Research Grant
1) Grant of Ministry of Health, Labor and Welfare Sick
House Syndrome(SHS)---Studies of patho-physiology,
method of diagnosis and its treatment.
2) Grant of Ministry of Health, Labor and Welfare Eating
disorder- Studies of neurology and endocrinology.
3) Grant of Ministry of Health, Labor and Welfare A
guideline for the diagnosis and treatment of psychosomatic diseases 2002
4) Grant of Ministry of Health and Welfare Studies ofstress management

Select Publications
1. Suematsu H, Ishikawa H, Kuboki T, Ito T. Statisticalstudies on anorexia nervosa in Japan: detailed clinicaldata on 1,011 patients. Psychother Psychosom43(2):96-103, 1985.
2. Suematsu H, Kuboki T, Itoh T. Statistical studies on
the prognosis of anorexia nervosa. Psychother
Psychosom 43(2):104-12, 1985.
3. Suematsu H, Kuboki T, Ogata E. Anorexia nervosa inmonozygotic twins. Psychother Psychosom 45(1):4650, 1986.
4. Kuboki T, Suematsu H, Ogata E, Yamamoto M,
Shizume K. Two cases of anorexia nervosa associatedwith Graves’ disease. Endocrinol Jpn 34(1):9-12, 1987.
5. Kumano H, Kuboki T, Tawara R, Sasaki T, Nomura S,
Suematsu H, Ogata E. Interrelationship between
serum muscle enzymes and a low T3 in anorexia nervosa. Endocrinol Jpn 37(4):583-9, 1990.
6. Kuboki T, Ide M, Kumano H, Nomura S, Suematsu H.
Historical review of panic disorder and clinical studies
in Japan． Journal of Psychiatric Research24(Suppl.1):70-72, 1990.
7. Kuboki T, Nomura S, Wada M, Akabayashi A,
Nagataki M, Suematsu H, Yokoyama K, Araki S.
Multidimensional assessment of mental state in occupational health care--combined application of threequestionnaires: Tokyo University Egogram (TEG),
Time Structuring Scale (TSS), and Profile of Mood
States (POMS). Environ Res 61(2):285-98, 1993.
8. Nakamoto C, Kumano H, Horie H, Nomura S, Kuboki
T, Suematsu H, Yasushi M, Saitou S, Chijiiwa M.
Psychological and physiological effects of photic feedback system. Shinshin-Igaku 33(5):371-80, 1993.

185

9. Horie H, Kumano H, Nomura S, Kuboki T, Suematsu
H, Hagura R. Multiple psychosocial factors influencing diabetes mellites: Multivariate analysis by quantification type I method. Shinshin-Igaku 33(8):667674, 1993.
10. Berger D, Saito S, Ono Y, Tezuka I, Shirahase J,
Kuboki T, Suematsu H. Dissociation and child abuse
histories in an eating disorder cohort in Japan. Acta
Psychiatr Scand 90(4):274-80, 1994.
11. Berger D, Ono Y, Saito S, Tezuka I, Takahashi Y, Uno
M, Ishikawa Y, Kuboki T, Asai M, Suematsu H.
Relationship of parental bonding to child abuse anddissociation in eating disorders in Japan. Acta
Psychiatr Scand 91(4):278-82, 1995.
12. Kumano H, Kuboki T, Ide M, Okabe F, Suematsu H.
Exercise tolerance evaluated by respiratory gas analysis during treadmill exercise test in panic disorderpatients．Jpn J Psychosom Med 35(8):671-679, 1995.
13. Horie H, Kumano H, Kuboki T, Suematsu H, Hagura
R. Multiple psychosocial factors influencing noninsulin-dependent diabetes mellites (II). ShinshinIgaku 35(6):501-509, 1995.
14. Kuboki T, Nomura S, Ide M, Suematsu H, Araki S.
Epidemiological data on anorexia nervosa in Japan.
Psychiatry Res 16;62(1):11-6, 1996.
15. Kumano H, Horie H, Shidara T, Kuboki T, Suematsu
H, Yasushi M. Treatment of a depressive disorderpatient with EEG-driven photic stimulation.
Biofeedback Self Regul 21(4):323-34, 1996.
16. Kumano H, Yamauchi Y, Matsumoto S, Sakano Y,
Kuboki T, Suematsu H. Cognitive behavior therapy
for eating disorders: Its advantages and shortcomings. Jpn J Psychosom Med 37(1):55-60, 1997.
17. Yoshiuchi K, Nomura S, Ando K, Ohtake T,
Shimosawa T, Kumano H, Kuboki T, Suematsu H,
Fujita T. Hemodynamic and endocrine responsiveness to mental arithmetic task and mirror drawing
test in patients with essential hypertension. Am J
Hypertens 10(3):243-9, 1997.
18. Nakao M, Nomura S, Shimosawa T, Yoshiuchi K,
Kumano H, Kuboki T, Suematsu H, Fujita T. Clinicaleffects of blood pressure biofeedback treatment on
hypertension by auto-shaping. Psychosom Med
59(3):331-8, 1997.
19. Kumano H, Horie H, Kuboki T, Suematsu H, Sato H,
Yasushi M, Kamei T, Masumura S. EEG-driven photic
stimulation effect on plasma cortisol and beta-endorphin. Appl Psychophysiol Biofeedback 22(3):193-208,
1997.
20. Nakao M, Nomura S, Yamanaka G, Kumano H,
Kuboki T. Assessment of patients by DSM-III-R and
DSM-IV in a Japanese psychosomatic clinic.
Psychother Psychosom 67(1):43-9, 1998.
21. Nakao M, Shimosawa T, Nomura S, Kuboki T, Fujita
T, Murata K. Mental arithmetic is a useful diagnostic
evaluation in white coat hypertension. Am J
Hypertens 11(1 Pt 1):41-5, 1998.
22. Yoshiuchi K, Kumano H, Nomura S, Yoshimura H,
Ito K, Kanaji Y, Ohashi Y, Kuboki T, Suematsu H.
Stressful life events and smoking were associated186with Graves’ disease in women, but not in men.
Psychosom Med 60(2):182-5, 1998.
23. Nakao M, Kumano H, Nomura S, Kuboki T,
Yokoyama K, Murata K. Fatigue mood as an indicator discriminating between anorexia nervosa andbulimia nervosa. Acta Psychiatr Scand 97(3):202-5,
1998.
24. Yoshiuchi K, Kumano H, Nomura S, Yoshimura H,
Ito K, Kanaji Y, Kuboki T, Suematsu H. Psychosocial
factors influencing the short-term outcome ofantithyroid drug therapy in Graves’ disease.
Psychosom Med 60(5):592-6, 1998.
25. Nakao M, Kumano H, Nomura S, Kuboki T, Murata
K. Assessment of ego state in anorexia nervosa andbulimia nervosa. Acta Psychiatr Scand 99(5):385-7,
1999.
26. Nakao M, Nomura S, Shimosawa T, Fujita T, Kuboki
T. Blood pressure biofeedback treatment, organ damage and sympathetic activity in mild hypertension.
Psychother Psychosom 68(6):341-7, 1999.
27. Fukunishi I, Berger D, Wogan J, Kuboki T.
Alexithymic traits as predictors of difficulties withadjustment in an outpatient cohort of expatriates inTokyo. Psychol Rep 85(1):67-77, 1999.
28. Kumano H, Orii Y, Suzukamo Y, Yamauchi Y,
Munakata M, Yoshinaga K, Seto M, Sakano Y, AgariI, Kuboki T. A trial of developing the Japanese briefversion of the Short Interpersonal Reactions
Inventory: To measure the Cancer-prone Personality.
Jpn J Psychosom Med 39(5):55-60, 1999.
29. Nakao M, Nomura S, Shimosawa T, Fujita T, Kuboki
T. Blood pressure biofeedback treatment of whitecoat hypertension. J Psychosom Res 48(2):161-9,
2000.
30. Nakahara R, Yoshiuchi K, Yamanaka G, Sasaki T,
Suematsu H, Kuboki T. Coping skills in Japanesewomen with eating disorders. Psychol Rep 87(3 Pt
1):741-6, 2000.
31. Kumano H, Orii Y, Yamauchi Y, Seto M, Agari I,
Sakano Y, Munakata M, Yoshinaga K, Sasaki T,
Kuboki T. A trial of developing the Japanese briefversion of the Short Interpersonal Reactions
Inventory (2nd report): Revision to the 33 items version. Jpn J Psychosom Med 40(6):447-54, 2000.
32. Nakao M, Yamanaka G, Kuboki T. Major depression
and somatic symptoms in a mind/body medicine
clinic. Psychopathology 34(5):230-5, 2001
33. Kumano H, Kuboki T, Orii Y, Fukuse T, Hirata T,
Shinohara K, Seto M, Agari I, Sakano Y. The investigation of the discriminant validity of the measurement of type C personality using the Japanese briefversion of the Short Interpersonal Reactions
Inventory (SIRI33). Jpn J Psychosom Med 41(8):59399, 2001.
34. Tsujiuchi T, Kumano H, Yoshiuchi K, He D, Tsujiuchi
Y, Kuboki T, Suematsu H, Hirao K. The effect of Qigong relaxation exercise on the control of type 2 diabetes mellitus: a randomized controlled trial.
Diabetes Care 25(1):241-2, 2002.

35. Nakao M, Barsky AJ, Kumano H, Kuboki T.
Relationship between somatosensory amplification
and alexithymia in a Japanese psychosomatic clinic.
Psychosomatics 43(1):55-60, 2002.
36. Nakao M, Yamanaka G, Kuboki T. Suicidal ideationand somatic symptoms of patients with mind/body
distress in a Japanese psychosomatic clinic. SuicideLife Threat Behav 32(1):80-90, 2002.
37. Nakao M, Ando K, Nomura S, Kuboki T, Uehara Y,
Toyooka T, Fujita T. Depressive mood accompanies
hypercholesterolemia in young Japanese adults. Jpn
Heart J 42(6):739-48, 2001.

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Department of ClinicalLaboratory Medicine
Outline and Research Objectives
Department of Clinical Laboratory Medicine, Tokyo University Graduate School of
Medicine, was established in 1997 following foundation of Department of Laboratory
Medicine, Faculty of Medicine, University of Tokyo in 1984. We have also ClinicalLaboratory Center which was established in 1952 in Tokyo University Hospital. The
Department cooperates with Clinical Laboratory Center not only in giving lectures and clinical education to medical students but also in carrying out researches. The Department ofClinical Laboratory Medicine is composed of only two staffs, a professor and an associate
professor, but Clinical Laboratory Center possesses four positions of lecturers and two of
associates. The professor and the associate professor of the Department are the director andthe vice-director of Clinical Laboratory Center, respectively.
In our department five or six research projects have been usually undertaken.

Faculties and Students
Professor and Chair Kazuhiko Nakahara, M.D. (1997~)
Associate Professor Kiyoshi Kitamura, M.D.(1998~2002)
Graduate Student....................1
Secretary .................................1

Past Research and Major Accomplishments
1. Cell surface analysis using flow cytometry
The major purpose of this project was to apply the
immunological methods to actual clinical fields, forinstance to diagnosis hematological malignancies suchas leukemia and lymphoma. We found that a kind of
glycolipids, asialo GM1, is the candidate of cell surface
markers for diagnosis of acute lymphoblastic
leukemia by using the method of flow cytometry. For
the next steps in this project, we discovered flow
cytometric analysis using several monoclonal antibodies reacted against human blood cells, especially lymphocytes, is very useful tool to diagnose humanhematological disorders. To give an example, we mentioned that adult T-cell leukemia cells are CD4 positive but negative for CD8, and very clear pattern ofthe positive sharp peak could be shown on the display of flow cytometry. We also demonstrated that
not only single-color but also multi-color analysis
with monoclonal antibodies and flow cytometry isvery important to investigate some kinds of illness inmore detail.
For the purpose of evaluating lymphocyte activation, we have studied lymphocyte surface antigens,plasma soluble CD4 or CD8, and intracellular pH (pHi)of lymphocytes, especially in infectious mononucleo-

188

sis (IM) in which proliferation of T lymphocytes is
supposed to play an important role in regulation of EB
virus. Immunofluorescence analysis demonstrates
increase or decrease of lymphocyte subset as well as
expression of activation antigens. During IM, CD8+ T
cells are quantitatively the major population of
expanded lymphocytes and they coexpress HLA-DR
and CD45RO. Elevated serum levels of soluble CD8(sCD8) and soluble CD4 (sCD4) are regarded as markers of T cell activation. We demonstrated marked
increase in sCD8 and significant increase in sCD4
which strongly suggest activation of both CD8+ andCD4+. The elevation of plasma sCD8 is due to expansion of CD8+ subset as well as increased sCD8 releasefrom each CD8+ cell.
Additionally we tried to apply the flow-cytometricanalysis method to measure intracellular pH (pHi),
which shows one of the indices of cellular activation.
For the purpose of evaluating intracellular pH of eachlymphocyte subset, simultaneous measurement of
pHi and cell surface antigen was developed using 2’-7’bis(carboxyethyl)5,6-carboxyfluorescein (BCECF) andflow-cytometry. A significant increase in pHi of CD8+lymphocytes from patients with IM and its correlation with number of CD8+-HLA-DR+ lymphocytes aswell as plasma levels of soluble CD8 strongly suggest
activation of CD8+ lymphocytes.
2. Effects of age, body-mass index and life-style
habits on clinical laboratory data.
2-1 Alcohol drinking and clinical laboratory data
Epidemiological studies have consistently shownthat light or light/moderate drinkers are at a lower
risk of coronary heart disease (CHD). The mecha-

nisms of this association include beneficial effects on
HDL- and LDL-cholesterol, insulin sensitivity, platelet
aggregation, blood coagulation and fibrinolysis.
However, drinking also has disadvantageous effects
on blood pressure, triglycerides and uric acid.
The purpose of our study was 1) to clarify whether
there are individual varieties regarding the responseof clinical laboratory data to alcohol consumption,and 2) what are the factors responsible for the individual varieties.
2-1-1 Are there individual varieties regarding the
response of clinical laboratory data to alcohol
consumption?
We performed a cross-sectional study of 3130
Japanese male workers with a body-mass index below
24. The subjects were divided into 2 groups; a normal
r-glutamyl transpeptidase (rGTP) (<40 IU/L) group anda high rGTP (>40 IU/L) group, and the values werecompared after adjusted for age, body-mass index,
exercise and smoking. The level of triglycerides
increased according to the amount of drinking in thehigh rGTP group, whereas no association wasobserved in the normal rGTP group. The level of HDLcholesterol increased with drinking in the normal andhigh rGTP groups, and no difference was observed in
the levels of HDL-cholesterol between the twogroups. The levels of uric acid and blood pressure also
increased with drinking in both groups, but theincrease was bigger in the high rGTP group than in
the normal rGTP group. The results indicated that
there was large individual variability in the responsesof the risk factors for coronary heart disease to drinking. Subjects whose rGTP responds less to drinking
may have less disadvantageous effects of drinking.
2-1-2 Is there any good way of drinking for health?
We performed a cross-sectional study of 3660 maleworkers to examine whether the weekly frequency ofdrinking affects clinical laboratory data when theweekly amount of drinking is the same. The results
suggested that the weekly frequency of drinking
might affect the levels of HDL-cholesterol independently of the weekly alcohol consumption.
2-1-3 Are there the genetic factors responsible forindividual varieties regarding the response ofclinical laboratory data to alcohol consumption?
We examined 133 male workers who drank >300g of alcohol per week to sudy the relationshipbetween clinical laboratory data and the genetic polymorphisms of alcohol dehydrogenase (ADH) 2 andaldehyde dehydrogenase (ALDH) 2. The results suggested that the ADH2 genotype influences the
responses of blood pressure, triglycerides, and uricacid to alcohol consumption, while the ALDH2 geno-

type influences the response of the erythrocyte meancell volume to alcohol consumption.
2-2 Others (HbA1C, proteinuria, leptin)
2-2-1 HbA1C
HbA1C has been suggested to deteriorate with age.
However, controversy exists as to whether this deterioration is a consequence of the aging process itself orof variables related to aging, such as concomitant disease, medication, body-mass index, physical activity,
and the change in distribution of body fat. We performed a cross-sectional survey of 7664 male Japanese
workers aged 20-59 years. This survey suggested that
HbA1C increased with age itself, and body-massindex and a hereditary predisposition to diabetes
affected this age-dependent increase in HbA1C.
2-2-2 proteinuria
We performed a cross-sectional survey of 5174
male Japanese workers. Blood pressure and a family
history of diabetes were independent factors associated with proteinuria in subjects with a HbA1C below
5.9% who were not under medication for diabetes. In
contrast, HbA1C, obesity and smoking were associated with proteinuria in subjects who were under medication for diabetes and/or have a HbA1C above 5.9%.
These findings suggest that maintaining a HbA1C
level below 5.9%, non-smoking and a standard bodyweight may reduce the prevalence of proteinuria inJapanese men. Healthy life–style habits and standard
body weight are especially important for subjects
with a family history of diabetes.
We also performed a cross-sectional study of 237
men to examine the relationship between proteinuria
determined with a reagent strip and the angiotensin
converting enzyme (ACE) genotypes. The results suggest that the ACE DD genotype may be an independent risk factor of proteinuria in Japanese men.
2-2-3 lepein
Leptin, the ob gene product secreted by adipocyte,
decreases food intake while it increases energy expenditure and functions as an important signal for the
regulation of body weight in mouse. To investigate
whether smoking affects the serum level of leptin, we
performed a cross-sectional study of 708 male workers. The findings suggested that among life-style
habits, physical activity, but not smoking or alcohol
consumption, significantly affects the serum level of
leptin in Japanese men.
The leptin receptor is an isoform of the B219 geneproduct, a member of the hematopoietin receptor
family, which is expressed in very primitive
hematopoietic cells. Recent studies showed that leptin
plus erythropoietin acted synergistically to increase

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erythroid development in vitro. These finding led us
to examine the relationship between the serum levelsof leptin and hemoglobin. The results of the above
cross-sectional study showed that the serum levels of
leptin is negatively associated with those of hemoglobin. Although the effect of leptin on hematopoiesis
may be modest, the results of our epidemiologic
study, together with the previous studies performedin vitro, suggest that leptin may play some role inhematopoiesis in humans.
3. Learning from patients at the points of biochemistry3-1 Study of apolipoprotein A-1 deficiency
We evaluated a 69-year-old Japanese woman withapolipoprotein (apo) A-I deficiency, high levels of
LDL-cholesterol, hypertension and impaired glucose
tolerance. The patient had corneal opacity, but neither
xanthomas, xanthelasma, nor tonsillar hypertrophy.
She was not symptomatic for coronary heart disease,and had normal electrocardiograms at rest and during
exercise on a cycle ergometer. She had severely
reduced levels of HDL-cholesterol (0.10-0.18 mmol/L)and no apo A-I (<0.6 mg/dL). LDL-cholesterol and apo
B as well as apo E were increased even under treatment with 10 mg pravastatin per day. Gel filtration
chromatography revealed that in addition to VLDL
and LDL fractions, she had apo A-II rich and apo E
rich fractions, which were present in the HDL fraction
separated by ultracentrifugation. A cytosine deletion
was identified by genomic DNA sequencing of the apo
A-I gene of the patient at the third base of codon 184
in the fourth exon, which led to a frame shift mutation and early termination at codon 200. This patientis the oldest among those with apo A-I deficiency
reported in the literature, and she had no symptomsof coronary heart disease despite the accumulated risk
for the disease.
3-2 Study of phytosterolemia
We diagnosed two women with multiple xanthomas, intermittent arthritis and thrombocytopenia
as phytosterolemia, an autosomal-recessive lipid storage disease, based on their increased serum concentrations of β-sitosterl, campesterol and sitostanol.
The gene responsible for this disease is located within
a distance of 18 cM between microssatellite markers
of D2S1788 and D2S1352 at chromosome 2p21. We
genotyped the patients and their family memberswith 16 microsatellite markers around this locus. The
results from the homozygosity mapping of one family
suggested that the gene was located within the distance of 12.6 cM between D2S2328 and D2S1352. We
have shortened the genetic distance by 5.4 cM.

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3-3 Another good effect of pioglitazone, aninsulin sensitizing drug
The findings obtained from two patients with diabetes and asthma suggested that pioglitazone may
ameliorate symptoms of asthma. Pioglitazone is oneof the thiazolidinedione compounds which have beenused as antidiabetic drugs. Recent studies haverevealed that thiazolidinedione compounds also havevarious nonhypoglycemic effects, such as antiinflamatory, antiatherosclerotic and anticancer effects, in cultured cells and in experimental animal models.
Elucidation of the mechanism may enable the designof a novel class of drugs for asthma.
4. Researches in physiological fields
4-1 Cardiac functions using ultrasoundMajor research interests are evaluation of left ventricular and left atrial functions, detection of tissue
ischemia using contrast echo, and assessment of cardiac state during inactivity.
4-2 Influence of diesel exhaust particles on respiratory functions
We reported that diesel exhaust particles (DEP)
induced nuclear factor-kappa B(NFκB) activation,
upregulated expression of intercellular adhesion molecule(ICAM)-1 in human bronchial epithelial cells, andbenzene-extracted components are important for the
major activity of DEP. Increased expression of transforming growth factor-β1 in small airway epithelium
was also demonstrated.
4-3 Investigations on brain functions using magnetoencephalography (MEG)
We installed the MEG system, which is a newly
developed tool for neurophysiology, and studied
somatosensory evoked magnetic fields to on- and offsets of rectangular pressure stimuli.

Current Research
1. Quantification of cell surface antigens using
flow cytometry
Conventional methods of flow cytometry provide
us only a positive percentage of cells. If we can measure and obtain cell surface antigen density, moredetail analysis must be possible. The purpose of thisproject is quantification of cell surface antigens withflow cytometry.
2. Analysis of Th1/Th2, Tc1/Tc2 lymphocytes
using flow cytometry
A method to analyze blood samples with EDTA-2K

was developed. Infectious diseases and immunocompromised patients are under investigation.

damage, especially renal damage, we are now investigating new marker in urine from various patients.

3. Relationship between genetic polymorphismsof alcohol metabolizing enzymes, and diabetic
control and the risk factors for CHD in patientswith diabetes.
Subjects: Non-drinkers and drinkers with diabetes
Methods: Genetic polymorphisms of ADH2 and
ALDH2 are examined by the PCR.
Objective:
1) Is the effect of alcohol drinking on diabetic control
associated with the genetic polymorphisms of
ADH2 and ALDH2?
2) Is the effect of alcohol drinking on the risk factors
for CHD associated with the genetic polymorphisms of ADH2 and ALDH2?
3) Are those effects observed only in drinkers andnot in non-drinkers?

7. Analysis of perfusion and function of the left
ventricule
Myocardial perfusion is assessed by myocardial
contrast echo while regional left ventricular function
is evaluated by strain data derived from myocardial
Doppler velocity information.

4. Identification of the gene mutation responsible
for phytosterolemia in our three patients.
Recent studies showed that phytosterolemia might
be explained by mutations of the genes that encode
new members of the ATP-binding cassette (ABC)transporter family, ABCG5 and ABCG8. Now we are
examining whether our patients have the gene mutations of ABCG5 and ABCG8.
5. Physiological roles of FGF-23
A new comer to our Department has cloned andidentified FGF-23 as a causative factors for tumorinduced hypophosphatemic rickets/osteomalacia. We
have also established an ELISA assay for biologically
active full-length FGF-23. We are now investigating
physiological roles of FGF-23 in phosphate and vitamin D metabolism.
6. Adrenomodullin
As another new comer participated in our
Department, this new project is started. He has been
studying the physiological role of vasoactive peptide
to regulate blood pressure by modulating sympathetic
tone and its possible role in preventing organ damage.
Adrenomedullin, a vasoactive peptide was discoveredfrom pheochromocytoma cells and it is known toexist not only adrenal cells but also vascular smoothmuscle cells, endothelial cells, and other organs. He
and his colleagues have generated adrenomedullinknockout mice and showed that adrenomedullin prevents organ damage by reducing oxidative stress. In
clinical study, they have shown adrenomedullin isbetter indicator of circulating blood volume than ANP
in patients under hemodialysis. The measurement ofblood adrenomedullin level will be clinically important marker. On the other hand, to diagnose organ

8. Relationship between respiratory function andvarious pathophysiological condition
The influence of sleep apnea syndrome, lung cancer, asthma, aging or respiratory rehabilitation on pulmonary function has been investigated by a memberof our Department.
9. Study using magnetoencephalography (MEG)
We have not only been making efforts to obtainreimbursement for clinical MEG studies but also beenorganizing several research groups which consists ofmedical doctors from the department of neurology,
physiology, neurosurgery, pediatrics and otolaryngology in the University Hospital and researchers from
University of Tokyo School of Medicine. Current
themes are follows.
1) Epilepsy evaluation-spike source localization
2) Pre-surgical functional mapping-somatosensory,
motor, auditory, visual and language
3) Auditory evoked magnetic response as a measureof sensory gating in schizophrenia
4) Abnormality of mismatch negativity in schizophrenic patients
PromoteFuture Prospects
We have a future planning in each research theme
as follows.
1) Apply the quantity analysis method of cell surface
antigens with flow cytometry to actual clinicalsamples and evaluate its usefulness.
2) Monitor immunological status of immunocompromised patients and patients with infectious diseases by using lymphocyte activation markers.
3) Prospective study on the relation between the mortality due to CHD and alcohol drinking, whichtakes genetic polymorphisms of alcohol metabolizing enzymes into consideration4) Elucidation of the structures and functions of
ABCG5 and ABCG8 and design of a novel class ofdrugs for hypercholesterolemia5) Study on physiological and pathophysiologocalroles of FGF-23

191

6) Find new markers to diagnose early phase of renal
damage in diabetes or hypertensive patients aswell as to diagnose response to treatment.
7) Three dimensional analysis of left ventricular perfusion and function using cardiac ultrasonography8) Promote EEG and MEG studies of higher cognitive
brain function, especially focused on the investigation of gamma-band activities which are thought
to play a key roll in perception and cognition.
It might be said that the research themes of ourdepartment are diverse and contain a wide range ofsubjects. Although we have actually a lot of research
projects simultaneously, we can say that it is the characteristic of our department and our common themeis laboratory examination and the key word is laboratory medicine. The purpose of our research is to apply
our research products to actual clinical fields, and contribute to human health and welfare. But today’s
problem in our department is the shortage of thenumber of graduate students. We must make an effort
to obtain them.

Research Grants
1. Grant-in-Aid for Scientific Research (B) from The
Ministry of Education, Science, Sports and Culture(1998~2000) : The new application of flow cytometry – Quantification of cell surface antigens and its
clinical application –2. Grant-in-Aid for Scientific Research (B) from The
Ministry of Education, Science, Sports and Culture(2001~2003) : Practical utilization of selective cell
information using flow cytometry and its clinicalapplication
3. Grant-in-Aid for Scientific Research (C) from The
Ministry of Education, Science, Sports andCulture(1999/2000) : Studies on phytosterolemia:
Identification of the pathologic gene and examination of its pathogenesis
4. Grant-in-Aid for Scientific Research (C) from The
Ministry of Education, Science, Sports and Culture(2001/2002) : Reverse cholesterol transport inapolipoprotein A1 deficiency
5. Grants-in-aid for Promotion of AIDS Research from
the Ministry of Health and Welfare of Japan(2000~2002)

Select Publications
1. Ｎakahara, K., Ohashi, T., Oda, T., Hirano, T., Kasai,
M., Okumura, Ｋ.＆ Tada, T. : Asialo GM１as a cellsurface marker detected in acute lymphoblastic
leukemia. New England J Medicine. 302：674-677,
1980.
2. Ｎakahara, Ｋ., Ohashi, T., Ichimaru M., Amagasaki,T., Shimoyama, M., Nakauchil ＆ kumura, K. :Analyses of adult T-cell leukemia using the mono-

192

clonal (anti-Leu-l, anti-Leu-2a and anti-Leu-3a) andheterologous anti-glycolipid (anti-asialo GML) antlbodies. Clinical Immunology and Imunopathology,
25：43-52, 1982.
3. Yoneyama, A., Nakahara, K., Higashihara, M. ＆Kurokawa : Increased levels of soluble CD8 and CD4
inpatients with infectious mononucleosis. Brit J
Haematol 89 : 47-54, 1995.
4. Aoki, K., Suzuki, K., Sugano, A., Tasaka, T., Nakahara,K., Kuge, O. Omori, A. ＆ Kasai, M. : A novel gene,Translin, encodes a recombination hotspott binding
protein associated with chromosomal translation.
Nature Genetics lO: 167-174, 1995.
5. Aoki, I., Homori, M., Nakahara, K., Higashi, K. ＆Ishikawa : Effects of recombinant humaninterleukin-1 (rhIL-), -1 (rhIL-1) and rhIL-1 receptor
antagonist (rhIL1ra) on erythroid progenitors(CFU-E
and BFU-E) in human bone marrow. Exp
Hematology 23(3):217-25, 1995.
6. Mashige, F., Matsushima, Y., Miyata, C., Yamada, R.,
Kanazawa, H. Sakuma, I., Takai, N., Shinozuka, N.,
Ohkubo, A and Nakahara, K. : Simultaneous determination of catecholamines, their basic metabolites
and serotonin in urine by high-perfomance liquid
chromatography using a mixed-mode column and aneight-channel electrochemical detector. Biomedical
Chromatography. 9 : 221-225, 1995.
7. Watanabe, T., Matsushima, S., Okazaki, M.,
Nagamatsu, S., Hirosawa, K., Uchimura, H. andNakahara, K. : Localization and ontogeny of GLUT3
expression in the rat retina. Developmental Brain
Research. 94, 60-66, 1996.
8. Watanabe, T., J. T. Voyvodic., T. Chan-Ling., Sagara,
H., Hirosawa, K., Mio, Y., Matsushima, S., Uchimura,H., Nakahara, K. and Martin Raff: Differentiationand morphogenesis in pellet culture of developing rat
retinal cells. The Jounal of Comparative Neurology.
377: 341-350, 1997.
9. Hashimoto Y, Togo M, Sato H, Hashimoto N,
Watanabe T, Kurokawa K, and Nakahara K.
Characteristics of protein kinase C-independent exocytosis in human platelets. Thromb Res 88: 51-58,
1997
10. Aoki, K. Inazawa, J., Takahashi, T., Nakahara, K.,
and Kasai, M. : Genomic structure and chromosomal
localization of the gene encoding translin, a recombination hotspot bindlng protein. Genomics, 43 (2) :237-241, 1997.
11. Hashimoto,Y., Togo,M., Sato,H., Hashimoto,N.,
Watanabe,T., Kurokawa,K. and Nakahara,K. :Characteristics of protein kinase C-dependent exocytosis in human platelets. Thrombosis Research, 88(1) : 51-58, 1997.
12. Sakurai, S., Shiojima, I., Tanigawa, T., and Nakahara,
K. : Aminoglycosides prevent and dissociate theaggregation of platelets in patients with EDTAdependent pseudothrombocytopenia. British J. of
Haematology,99: 817-823,1997.
13. Koike, Y., Yoneyama, A., Shirai, J., Ishida, T., Shoda,E., Miyazaki, K., Sunaga, S., Horie, R., Aoki, K.,Koike, K., Ogata, I., Takahara, T., Kato, T., Nakahara,K., Kariya, T., and Higashihara, M. : Evaluation ofthrombopoiesis in thrombocytopenic disorders by
simultaneous measurement of reticulated platelets ofwhole blood and serum thrombopoietin concentrations. Thromb Haemost. 79:1106-1110,1998.
14. Kawasaki, S., Takizawa, H., Ohtoshi, T., Takeuchi,N., Kohyama, T., Nakamura, H., Asama, T.,
Kobayashi,K., Nakahara, K., Morita, Y. And
Yamamoto, K. : Roxithromycin in hibits cytokine
production by and neutrophil attachment to humanbronchial epithelial cells in vitro. American Agentsand Chemotherapy. 42 (6) 1499-1502, 1998.
15. Yan, K., Nakahara, K., Awa, S., Nishibori, Y.,
Nakajima, N., Kataoka, S. Maeda, M., Watanabe, T.,
Matsushima, S., and Watanabe, N. : The increase ofmemory T cell subsets in children with idiopathic
nephrotic syndrome. Nephron.79, 274-27８, 1998.
16. Aoki, K., Meng, G., Suzuki, K., Takashi, T., Kameoka,
Y., Nakahara, K., Ishida, R., Kasai, M. : RP58 associates with condensed chromatin and mediates asequence-specific ranscriptional repression. The
Journal of Biological Chemistry. 273 (41), Issue ofOctober 9, 26698-26704, 1998.
17. Hashimoto, Y., Hutamura, A., Watanabe, N., Togo,
M., Sato, H., Hara, M., Tsukamoto, K., Kimura, S.,
and Nakahara, K. : Relationship between glcosylated
hemoglobin and the prevalence of proteinuria inJapanese men. Internal Medicine, 38 (1), 6-11,1999.
18. Hashimoto, Y., Futamura, A., Watanabe, N., Togo,
M., Sato, H., Hara, M., Tsukamoto, K., imura, S., andNakahara, K. : Relationship between glycosylated
hemoglobin and the prevalence of roteinuria inJapanese men. Intern Med., 38(1):6-11, 1999.
19.Togo, M., Tsukamoto, K., Satoh, H., Hara, M.,
Futamura, A., Nakarai, H, Nakahara, K., Hashimoto,Y. : Relationship Between Levels of Leptin and
Hemoglobin in Japanese Men. Blood., 93:4444-4445,
1999.
20. Okubo, S., Mashige, F., Hashimoto, Y., Nakahara, K.,
Kanazawa, H., Matsushima, Y., Fukushima, T.,
Huang, Y., and Imai,I.;D-Serine concentration inblood serum of liver transplanted patient.
Chromatography, 21:43-47,2000.
21. Takizawa, H., Abe, S., Ohtoshi, T., Kawasaki, S.,
Takami, K., Desaki, M., Sugawara, I., Hashimoto, S.,
Azuma, A., Nakahara, K. and Kudoh, S. : Diesel
exhaust particles up-regulate expression of intercellular adhesion molecule-1 (ICAM-1) in human epithelial cells. Clin Exp Immunol 120: 356-362, 2000.
22. Meng, G., Inazawa, J., Ishida, R., Tokura, K.,
Nakahara, K., Aoki, K. & Kasai, M. : Structural analysis of the gene encoding RP58, a sequence-specifictransrepressor associated with heterochromatin.
Gene 242, 59-64, 2000.
23. Meng, G., Aoki, K., Tokura, K., Nakahara, K.,
Inazawa, Johji. & Kasai, M. : Genomic structure andchromosomal location of the gene encoding TRAX, a
Translin-associated factor X. J Hum Genet 45: 305308, 2000.

24. Okubo, S., Mashige, F., Omori, M., Hashimoto, Y.,
Nakahara, K., Kanazawa, H. & Matsushima, Y. :Enantiomeric determination of L-and D-lactic acid inhuman cerebrospinal fluid by chiral ligand exchange
high-performance liquid chromatography. Biomed
Chromotogr 14: 474-477, 2000.
25. Watanabe, T. and Nakahara, K. : Quantification ofcell surface molecule density by flow cytometry.
Symposium Abstract, The Sixth Asian Conference ofClinical Pathology. S89, 2000.
26. Yumoto, M., Saitoh, O., Kaneko, Y., Anami, K.,
Matsuda, H., Ugawa, Y. and Nakahara, K. :Somotosensory evoked magnetic fields to on-and offsets of rectangular pressure stimuli. Proc. of 12th
International Conference on Biomagnetism,243-246,
2000.
27. Togo, M., Hashimoto, Y., Futamura, A., Tsukamoto,K., Satoh, H., Watanabe, T., Nakarai, H., Nakahara,
K.,and Kimura, S. : Relationship between serum levelof leptin and life-style habits in Japanese men. Horm
Res 54,169-173,2000.
28. Togo, M., Toda, T., L.A.Nguyen, Kubota, S.,
Tsukamoto, K., Satoh, H., Hara, M., Iso-o,N., Noto,
H., Kimura, S., Nakahara, K., Seyama, Y. andHashimoto, Y. : Genetic analysis of phytosterolaemia. J. Inherit. Metab. Dis. 24, 43-50, 2001.
29. Sakurai,S., Takenaka,K., Shiojima, I., Watanabe,F.,
Sonoda, M., Uno,K., Kuwada,Y., Nakahara,K., andNagai,R.: Traumatic Tricuspid Insufficiency.
Echocardiography:A J. of CV Ultrasound & Allied
Tech. 18 (4) 303-304, 2001.
30. Kawasaki, S., Takizawa, H., Takami, K., Desaki, M.,
Okazaki, H., Kasama, T., Kobayashi, K., KYamamoto,K., Nakahara, K., Tanaka, M., Sagai, M., andOhtoshi, T. : Benzene-extracted components areimportant for the major activity of diesel exhaust
particles (DEP): Effect on IL-8 gene expression inhuman bronchial epithelial cells. Am J Repir Cell Mol
Biol 24, 419-426, 2001.
31. Hashimoto, Y., Futamura, A., Nakarai, H., Yokota,
H., Omura, M., Tsukamoto, K., Togo, M., Sato, H.,
Hara, M., Isoo, N., Taguchi, J., and Nakahara, K. :Association between deletion polymorphism ofangiotensin converting enzyme gene and proteinuria
in Japanese overweight men., J Occup Health.43,8084, 2001.
32. Sakurai, S., Takenaka, K., Nakahara, K., and Nagai,
R. : Traumatic tricuspid insufficiency.
Echocardiography 18 : 303, 2001.
33. Takizawa, H., Tanaka, M., Takami, K., Ohtoshi, T.,
Ito, K., Satoh, M., Okada, Y., Yamasawa, F., Umeda,
A., and Nakahara, K. : Increased Expression ofTransforming Growth Factor-β1 in Small Airway
Epithelium from Tobacco Smokers and Patients with
Chronic Obstructive Pulmonary Disease (COPD). Am
J Respir Crit Care Med 163, 1476-1483, 2001.
34. Mashige. F., Tsuno H. N., Maekawa, M., Okada, S.,
Tanaka, N., Matsui, S., Watanabe, S. and Nakahara,
K. : Noninvasive combination assay of free L-fucose
and suflated bile acids in urine for diagnosis of liver193

and pancreatic diseases and solid tumor. Journal of
Analytical Bio-Science 24(4) 317-323, 2001.
35. Hashimoto,Y., Futamura, A., Nakarai, H., andNakahara, K. : Relationship between response of glutamly transpetidase to alcohol drinking and risk
factors for coronary heart disease. Athreosclerosis
158 465-470, 2001.
36. Sakurai, S., Takenaka, K., Shiojima I., Watanabe, F.,
Onoda, M., Uno, K., Kuwada,Y., Nakahara, K., andNagai, R. : Traumatic Tricuspid Insufficiency.
Echocardiography:A J. of CV Ultrasound & Allied
Tech. 18 (4) 303-304, 2001.
37. Kawasaki, S., Takizawa, H., Takami, K., Desaki, M.,
Okazaki, H., Kasama, T., Kobayashi, K., Yamamoto,K., Nakahara, K., Tanaka, M., Sagai, M., andOhtoshi, T. : Benzene-extracted components areimportant for the major activity of diesel exhaust
particles : Effect on interleukin-8 gene expression inhuman bronchial epithelial cells. Am J Respir Cell
Mol Biol. 24, 419-426, 2001.
38. Hashimoto, Y., Futamura, A., Nakarai, H., andNakahara, K. : Effects of the frequency of alcohol
intake on risk factors for coronary heart disease.
European Journal of Epidemiology 17, 307-312, 2001.
39. Sakatani, T., Shimazaki, C., Hirai, H., Okano, A.,
Hatsuse, M., Okamoto, A., Takahashi, R., Ashihara,E., Inaba, T., Yokota, H., Nakahara, K., Hirai, H. andNakagawa, M. : Early relapse after hgh-dose
chemotherapy rescued by tumor-free autologous
peripheral blood stem cells in acute lymphoblastic
leukemia : Importance of monitoring for WT1mRNA quantitatively. Leukemia and Lymphoma42(1-2), 225-229, 2001.
40. Yokota, H., Tsuno, N. H., Tanaka, Y., Fukui, T.,
Kitamura, K., Hirai, H., Osumi, K., Itou, N., H. Satoh,
M. Okabe, S, Nakahara, K. : Quantification of
Minimal Residual Disease in Patients with ela2 BCRABL Positive Acute Lymphoblastic Leukemia Using a
Real-Time RT-PCR Assay. Leukemia(16), 1167-1175,
2002.
41. Yokota H, Hashimoto Y, Okubo S, Yumoto M,
Mashige F, Kawamura M, Kotani K, Usuki Y,
Shimada S, Kitamura K, Nakahara, K. Apolipoprotein
A-I deficiency with accumulated risk for CHD but nosymptoms of CHD. Atherosclerosis 162: 399-407,
2002
42. Hashimoto, Y., and Nakahara, K. : Improve of asthma after administration of pioglitazone. Diabetes
Care, 25(2),401, 2002.
43. Kanazawa, H., Okada, A., Matsushima, Y., Yokota,
H., Okubo, S., Mashige, F., and Nakahara, K. :Determination of omeprazole and its metabolites inhuman plasma by liquid chromatograpy-mass spectrometry. Journal of Chromatgrahy A, 949, 1-9, 2002.
44. Ono, Y., Mashige, F., Sano, S., Nanba, Y., Okubo, S.,
Hashimoto, Y., and Nakahara, K. : α 1Antichymotrypsin in serum and cerebrospinal fluid
Part1 : Development of latex agglutination test for
determination of α 1-antichymotrypsin. Journal of
Analytical Bio-Science, 25(2), 153-158, 2002.

194

45. Mashige, F., Ono, Y., Nanba, Y., Okubo, S.,
Hashimoto Y., and Nakahara, K. : α 1Antichymotrypsin in serum and cerebrospinal fluid
Part2 : Clinical implications of α 1-antichymotrypsin
and other acute phase reactant proteins in
Alzheimer-type dementia and other diseases. Journalof Analytical Bio-Science, 25(2), 159-163, 2002.
46. Kitamura, K., Miwa, S., Ikemoto, T., Tatsumi, N.,
Nakahara, K., Higashi, K., and Watanabe, K. :Proposal of guidelines for surface antigen analysis ofperipheral lymphocytes using flow cytometry andpreliminary quality control. Global Standardization
and Advanced quality Management’01, Eibun Press,
Ltd., Osaka, Japan, pp48, 2002.
47. Hashimoto,Y., Nakayama, T., Futamura, A., Omura
M., and Nakahara, K. : Erythrocytes mean cell volume and genetic polymoprphism of aldehyde dehydrogenase 2 in alcohol drinkers. Blood 99(9):3487-8,
2002.
48. Hashimoto, Y., Nakayama, T., Futamura, A., Omura,M., Nakarai, H., and Nakahara, K. : Relationshipbetween genetic polymorphisms of alcohol-metabolizing enzymes and changes in risk factors for coronary heart disease associated with alcohol consumption. Clinical Chemistry, 48(7), 1043-1048, 2002.
49. Takami, K., Takuwa, N., Okazaki, H., Kobayashi, M.,
Ohtoshi, T., Kawasaki, S., Dohi, M., Yamamoto, K.,
Nakamura, T., Tanaka, M., Nakahara, K., Takuwa,
Y., and Takizawa, H. : Interferon-gamma Inhibits
hepatocyte growth factor-stimulated cell proliferation of human bronchial epithelial cells upregulationof p27kip1 cyclin-dependent kinase inhibitor.
American Journal of Respiratory Cell & Molecular
Biology, 26(2), 231-238, 2002
50. Takenaka, K., Suzuki, Y., Uno, K., Sato, M., Komuro,T., Haruna Y., Kobayashi, H., Kawakubo, K., Sonoda,
M., Asakawa, M., Nakahara, K., and Gunji, A. :Effects of rapid saline infusion on orthostatic intolrerance and automatic tone after 20 days bed rest.
Am J Cardiol 89, 557-561, 2002.

Department ofTransfusion Medicine
Outline and Research Objectives
The Department of Transfusion Medicine was established in 1949, and actually is composed by 4 medical doctors, 10 medical technicians, 1 nurse and 1 office assistant.

Faculties and Students
Professor and Chair

Yoichi Shibata, M.D., Ph.D. (sinceApril 1993)
Associate Professors absent
Lecturer
absentAssociate .................................3
Research Students...................4 research students fromthe department ofSurgical Oncology aredeveloping their research
projects at the department’s laboratory.
Secretary .................................1
Medical Technician...............10
Nurse.......................................1

4)

5)

6)

Past Research and Major Accomplishments
The major research projects developed at the
department are related to:1) Human platelet antigen/antibody detection andcharacterization: the mixed-passive hemagglutination (MPHA) assay, a serological method largely
used in Japan for the screening and characterization of platelet alloantigens and alloantibodies,
was developed at the department by Yoichi
Shibata. Using this method, a new platelet
alloantigen system, namely the HPA-4 system(Yuka/b), was discovered by Y. Shibata, et al.
2) Human endothelial cell antigen/antibody detection
and characterization: the MPHA method, developed by Y. Shibata, et al., was modified for thedetection and characterization of endothelial cellalloantigens and alloantibodies. Using thismethod, monoclonal antibodies reactive withtumor endothelial cells were developed at the
department. This monoclonals are being investigated for a possible application in cancer treatment.
3) Immunotherapy of cancer using LAK cells anddendritic cells: the effectiveness of lymphokineactivated killer (LAK) cells, as well as of peripheral
blood and monocyte-derived dendritic cells for
immunotherapy of cancer was investigated in

7)

8)

vitro, and were tested in a clinical trial for thetreatment of patients with gastrointestinal cancer
and brain tumors.
Development of new vaccines targeting the tumor
angiogenesis for the treatment of cancer: new vaccine strategies for the treatment of solid tumors
by targeting the tumor vasculature are being
developed at the department. The vaccine, tested
in an animal model of colon cancer metastasis,
showed to be very effective, and is being prepared
for the clinical trial.
Vaccination therapy of patients with habitualabortion using the husband’s lymphocytes, andinvestigation on their mechanism of action.
Effects of radiation and filtration on platelet function: the effects of radiation and the use of whiteblood cell (WBC)-reduction filters on the platelet
function (adhesion and aggregation) have beenstudied. Irradiation did not affect the platelet
function, but the WBC-reduction filters, depending on the type of filter, reduced the adhesive
capacity of platelets.
Development of a new methodology for the evaluation of platelet function: a new method, based on
the adhesive capacity of platelets, is being developed at the department.
Peripheral blood stem cells transplantation:
peripheral blood stem cells are isolated at the
department for transplantation in leukemic
patients.

Current Research
The research projects described above are stillongoing at the department of Transfusion Medicine,which are:1) Human platelet antigen/antibody detection andcharacterization, and study on their role in transfusion practice;2) Human endothelial cell antigen/antibody detection
and characterization, and study on their role in
the pathogenesis of inflammatory and autoimmune diseases, as well as in organ transplantation;3) Immunotherapy of cancer using LAK cells and dendritic cells;

1954) Development of new vaccines targeting the tumor
angiogenesis for the treatment of cancer;
5) Lymphocyte vaccination therapy for patients with
habitual abortion;
6) Effects of radiation and filtration on platelet function;7) Development of a new methodology for the evaluation of platelet function;
Peripheral blood stem cells transplantation.

Future Prospects
The future prospects of the department are as follows:
1) To develop the ideal method of platelet collection
and preservation for transfusion, especially related
to preservation of platelets without loss of theirfunction;
2) To develop a good methodology for the evaluationof platelet function;3) To develop a new therapeutic strategy to combat
cancer by targeting the angiogenic vasculature oftumors;4) To develop the vascular regeneration medicine
using peripheral blood and cord blood stem cells.

Research Grants(1) Grant-in-Aid for Scientific Research from the Ministryof Education, Culture, Sports, Science andTechnology of Japan –Kiban (C)(2); Development of a
new methodology for the detection of anti-endothelial cell antibodies and evaluation on their clinicalsignificance. 1997 – 1998;3 million yen.
(2) Grant-in-Aid for Scientific Research from the Ministryof Education, Culture, Sports, Science andTechnology of Japan (B)(2): Development of a monoclonal antibody-based strategy for cancer treatment
targeting the angiogenic vascular endothelium. 1999
– 2001;11,800 thousand yen.
(3) Grant-in-Aid for Scientific Research from the Ministryof Education, Culture, Sports, Science andTechnology of Japan- Grant for Highly-advanced
Medicine, Development of a cell-based and genebased strategies for the treatment of malignant andother life-threatening disorder of difficult control.
1998 – 2000; 131,304 thousand yen.

Select Publications
1. Osada T, Fujimaki T, Takamizawa M, Tsuno NH,
Kirino T, Shibata Y. Dendritic cells activate antitumor immunity for malignant intracranial germ celltumor: a case report. Japanese Journal of ClinicalOncology;31(8):403-6, 2001.
2. Osada T, Nagawa H, Kitayama J, Tsuno NH, Ishihara
S, Takamizawa M, Shibata Y. Peripheral blood dendritic cells, but not monocyte-derived dendritic cells,

196

can augment human NK cell function. Cell Immunol,;213(1):14-23, 2001.
3. Maejima M, Fujii T, Yamashita T, Hara N, Hamai Y,
Miki A, Kozuma S, Okai T, Shibata Y, Taketani Y.
Immunotherapy before and during pregnancy
improvespregnancy outcome in women who sufferfrom recurrent abortion and did not benefit from
immunotherapy before pregnancy. American Journalof Reproductive Immunology;39(1):12-5, 1998.
4. Maejima M, Fujii T, Okai T, Kozuma S, Shibata Y,
Taketani Y. Beta2-glycoprotein I-dependent anticardiolipin antibody in early recurrent spontaneousabortion. Human Reproduction;12(10):2140-2, 1997.
5. Maejima M, Fujii T, Kozuma S, Okai T, Shibata Y,
Taketani Y. Presence of HLA-G-expressing cells modulates the ability of peripheral blood mononuclear
cells to release cytokines. American Journal of
Reproductive Immunology;38(2):79-82, 1997.
6. Minchinton RM, Dawkins B, Chynoweth L, Pearson
H, Lown JA, Shibata Y. In pursuit of enigmatic
platelet antibodies--anti-HPA-2b and anti-HPA-3a.
Transfusion Medicine;6(3):289-91, 1996.
7. Takeuchi F, Kuwata S, Nakano K, Nabeta H, Hong
GH, Shibata Y, Tanimoto K, Ito K. Association of
TAP1 and TAP2 with systemic sclerosis in Japanese.
Clinical & Experimental Rheumatology;14(5):513-21,
1996.
8. Fujii T, Tsushima R, Okai T, Shibata Y, Taketani Y.
Allo-non-specific elevation of maternal killer cell
activity in intrauterine growth restriction.
International Journal of Gynaecology & Obstetrics;
52(3):237-42, 1996.
9. Chiu L, Nishimura M, Ishii Y, Nieda M, Maeshima
M, Takedani Y, Shibata Y, Tadokoro K, Juji T.
Enhancement of the expression of progesterone
receptor on progesterone-treated lymphocytes after
immunotherapy in unexplained recurrent spontaneous abortion. American Journal of Reproductive
Immunology;35(6):552-7, 1996.
10. Sunami E, Tsuno NH, Kitayama J, Osada T, Saito S,
Tomozawa S, Tsuruo T, Shibata Y, Nagawa H.
Decreased Synthesis of MMP-7 and Adhesion to
Extracellular Matrix Proteins of Human Colon Cancer
Cells Treated with Troglitazone. Surgery Today 2001
11. Tomozawa S, Tsuno NH, Sunami E, Hatano K,
Kitayama J, Osada T, Saito S, Tsuruo T, Shibata Y,
Nagawa H. Cyclooxygenase-2 overexpression correlates with tumour recurrence, especially haematogenous metastasis, of colorectal cancer. Br J Cancer;
83(3):324-328, 2000.
12. Sunami E, Tsuno N, Osada T, Saito S, Kitayama J,
Tomozawa S, Tsuruo T, Shibata Y, Muto T, Nagawa
H. MMP-1 is a prognostic marker for hematogenous
metastasis of colorectal cancer. Oncologist;5(2):10814, 2000.
13. Saito S, Tsuno N, Nagawa H, Sunami E, Zhengxi J,
Osada T, Kitayama J, Shibata Y, Tsuruo T, Muto T.
Expression of platelet-derived endothelial cell
growth factor correlates with good prognosis inpatients with colorectal carcinoma. Cancer Jan
1;88(1):42-9, 2000.
14. Tomozawa S, Nagawa H, Tsuno N, Hatano K, OsadaT, Kitayama J, Sunami E, Nita ME, Ishihara S, Yano
H, Tsuruo T, Shibata Y, Muto T. Inhibition of
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