Liposomes Part C

610 Pages·2003·6.515 MB·English

by  Colowick S.P., Kaplan N.O.

Preface
The origins of liposome research can be traced to the contributions by Alec
Bangham and colleagues in the mid 1960s. The description of lecithin disper-
sions as containing ‘‘spherulites composed of concentric lamellae’’ (A.D.
Bangham and R.W. Horne, J. Mol. Biol. 8, 660, 1964) was followed by the
observationthat‘‘thediffusionofunivalentcationsandanionsoutofspontan-
eouslyformedliquidcrystalsoflecithinisremarkablysimilartothediffusionof
such ions across biological membranes (A.D. Bangham, M.M. Standish and
J.C. Watkins, J. Mol. Biol. 13, 238, 1965). Following early studies on the
biophysicalcharacterizationofmultilamellarandunilamellarliposomes,inves-
tigators began to utilize liposomes as a well-defined model to understand the
structure and function of biological membranes. It was also recognized by
pioneersincludingGregoryGregoriadisandDemetriosPapahadjopoulosthat
liposomes could be used as drug delivery vehicles. It is gratifying that their
effortsandtheworkofthoseinspiredbythemhaveleadtothedevelopmentof
liposomalformulationsofdoxorubicin,daunorubicinandamphotericinBnow
utilizedintheclinic.Othermedicalapplicationsofliposomesincludetheiruse
as vaccine adjuvants and gene delivery vehicles, which are being explored in
thelaboratoryaswellasinclinicaltrials.Thefieldhasprogressedenormously
inthe38yearssince1965.
Thisvolumeincludesapplicationsofliposomesinimmunology,diagnostics,
and gene delivery and gene therapy. I hope that these chapters will facilitate
theworkofgraduatestudents,post-doctoralfellows,andestablishedscientists
entering liposome research. Other volumes in this series cover additional
subdisciplinesinliposomology.
The areas represented in this volume are by no means exhaustive. I have
tried to identify the experts in each area of liposome research, particularly
thosewhohavecontributedtothefieldoversometime.ItisunfortunatethatI
was unable to convince some prominent investigators to contribute to the
volume. Some invited contributors were not able to prepare their chapters,
despite generous extensions of time. In some cases I may have inadvertently
overlookedsomeexpertsinaparticulararea,andtotheseindividualsIextend
myapologies.Theirprimarycontributionstothefieldwill,nevertheless,notgo
unnoticed,inthecitationsinthesevolumesandintheheartsandmindsofthe
manyinvestigatorsinliposomeresearch.
xv
xvi preface
I would like to express my gratitude to all the colleagues who graciously
contributedtothesevolumes.IwouldliketothankShirleyLightofAcademic
Press for her encouragement for this project, and Noelle Gracy of Elsevier
Scienceforherhelpatthelaterstagesoftheproject.Iamespeciallythankfulto
my wife Diana Flasher for her understanding, support and love during the
seemingly never-ending editing process, and my children Avery and Maxine
fortheiruniquecuriosity,creativity,cheer,andlove.Finally,Iwishtodedicate
this volume to two other members of my family who have been influential in
my life, with their love and support since my childhood days, my aunt Sevim
UygurerandmybrotherDr.ArdaDu¨zgu¨nes,
Nejat Du¨zgu¨nes,
METHODS IN ENZYMOLOGY
EDITORS-IN-CHIEF
John N. Abelson Melvin I. Simon
DIVISIONOFBIOLOGY
CALIFORNIAINSTITUTEOFTECHNOLOGY
PASADENA,CALIFORNIA
FOUNDING EDITORS
Sidney P. Colowick and Nathan O. Kaplan
Contributors to Volume 373
Articlenumbersareinparenthesesandfollowingthenamesofcontributors.
Affiliationslistedarecurrent.
SalvadorF.Alin˜o(26),Departamentode ElenaBogdanenko(28),VNOrekhovich
Famacologia,FacultaddeMedicina,Uni- Institute of Biomedical Chemistry, Rus-
versidaddeValencia,AvdaBlascoIbanez sian Academy of Medical Sciences, 10,
15,46010Valencia,Spain Pogodinska ya Street, 119832 Moscow,
Russia
CarlR.Alving(2,3,10),Departmentof
Membrane Biochemistry, Walter Reed Jeff W.M. Bulte (12), Department of
ArmyInstituteofResearch,Washington, Radiology,TheJohnsHopkinsUniversity
D.C.20307 SchoolofMedicine,Baltimore,Maryland
21205
M. A. Arangoa (22), Department of
LauraBungener(5),DepartmentofMed-
Pharmacology and Pharmaceutical
ical Microbiology, Molecular Virology
Technology,SchoolofPharmacy,Univer-
Section, University of Groningen, 9713
sityofNavarra,31080Pamplona,Spain
AVGroningen,TheNetherlands
Udo Bakowsky (18), Department of Rolf Bunger (10), Department of
PharmaceuticalTechnologyandBiophar-
Membrane Biochemistry, Walter Reed
macy, University of Saarbruecken,
ArmyInstituteofResearch,Washington,
Germany
D.C.20307
Richard R.Bankert (33),Department of GerardoByk(23),LaboratoryofPeptido-
Microbiology, SUNY at Buffalo, 138 mimeticsandGeneticChemistry,BarIlan
Farber Hall, 3435 Main Street, Buffalo, University, Department of Chemistry,
NewYork14214 52900RamatGan,Israel
LajosBaranyi(10),DepartmentofMem- Jin-SooChang(9),MorganBiotechnology
brane Biochemistry, Walter Reed Army ResearchInstitute,341Pojung-Ri,Koon-
Institute of Research, Washington, D.C. sung-Myon, Youngin City, Kyonggi-Do
20307 449-910,SouthKorea
Marta Benet (26), Departamento de Fa- Myeong-Jun Choi (9), Charmzone
macologia,FacultaddeMedicina,Univer- Co.,Ltd., Bioimaterial Research Center,
sidaddeValencia,AvdaBlascoIbanez15, 301 Hankang Building, 184-11 Kwang-
46010Valencia,Spain jang-dong,Kwangjin-ju,Seoul,Korea
MichaelBodo(10),DepartmentofMem- Jaime Crespo (26), Departamento de Fa-
brane Biochemistry, Walter Reed Army macologia,FacultaddeMedicina,Univer-
Institute of Research, Washington, D.C. sidaddeValencia,AvdaBlascoIbanez15,
20307 46010Valencia,Spain
OttoC.Boerman(15),DepartmentofNu- ToosDaemen(5),DepartmentofMedical
clearMedicine(565),UniversityMedical Microbiology, Molecular Virology
Center Nijmegen, P.O. Box 9101, 6500 Section, University of Groningen, 9713
HBNijmegen,TheNetherlands AVGroningen,TheNetherlands
ix
x contributors to volume 373
Sumeet Dagar (13), Departments of StephenJ.Frost(16),DepartmentofClin-
Pharmaceutics and Pharmacodynamics, ical Biochemistry, The Princess Royal
University of Illinois at Chicago, 833 Hospital, Lewes Rd. Haywards Heath,
WoodStreet,Chicago,Illinois60612 WestSuxxesRH163LU,England
FranciscoDası´(26),DepartamentodeFa- M. Teresa Gira˜o Da Cruz (24), Depart-
macologia,FacultaddeMedicina,Univer- mentofBiochemistry,FacultyofSciences
sidaddeValencia,AvdaBlascoIbanez15, and Technology, University of Coimbra,
46010Valencia,Spain 3000Coimbra,Portugal
Robert J. Debs (34), Geraldine Brush Laurent Giraudo (7), Centre d’Immuno-
Cancer Research Institute, 2330 Clay logie de Marseille-Luminy, Campus de
Street,SanFrancisco,California94115 Luminy, Case 906, 13288 Marsielle
Cedex09,France
MarcelDeCuyper(12),Interdisciplinary
Research Center, Katholieke Universiteit Mitsuru Hashida (25), Graduate School
Leuven, Campus Kortrijk, B-8500 of Pharmaceutical Sciences, Kyoto Uni-
versity,Sakyo-ku,Kyoto606-850,Japan
Kortrijk,Belgium
C.TrosDeIlarduya(22),Departmentof Kazuya Hiraoka (30), Division of Gene
Therapy Science, Graduate School of
PharmacologyandPharmaceuticalTech-
Medicine, Osaka University, 2-2 Yama-
nology, School of Pharmacy, University
da-oka, Suita City, Osaka 565-0871,
ofNavarra,31080Pamplona,Spain
Japan
NejatDu¨zgu¨nes,(19,22,24,28),Depart-
ment of Microbiology, University of the DickHoekstra(18),DepartmentofMem-
brane Cell Biology, University of Gron-
PacificSchoolofDentistry,2155Webster
ingen, Antonius Deusinglaan 1, 9713 AV
Street,SanFrancisco,California94115
Groningen,TheNetherlands
Nejat K. Egˇilmez (33), Department of
LeafHuang(21),CenterforPharmacoge-
Microbiology, SUNY at Buffalo, 138
netics,SchoolofPharmacy,Universityof
Farber Hall, 3435 Main Street, Buffalo,
Pittsburgh, 633 Salk Hall, Pittsburgh,
NewYork14214
Pennsylvania15213
Abdelatif Elouahabi (20), Center for
AnkeHuckreide(5),DepartmentofMed-
Structural Biology and Bioinformatics,
ical Microbiology, Molecular Virology
CP 206/2, Campus Plaine-ULB, Blv du
Section, University of Groningen, 9713
Triomphe,1050Brussels,Belgium
AVGroningen,TheNetherlands
Henrique Faneca (19), Department of Yasufumi Kaneda (30), Division of Gene
Biochemistry, Faculty of Sciences and
Therapy Science, Graduate School of
Technology,UniversityofCoimbra,3000
Medicine,OsakaUniversity,2-2Yamada-
Coimbra,Portugal
oka,SuitaCity,Osaka565-0871,Japan
SylviaFong(34),GeraldineBrushCancer Shigeru Kawakami (25), Faculty of
ResearchInstitute,2330ClayStreet,San
Pharmaceutical Sciences, Nagasaki
Francisco,California94115
University,Magaski852-8521,Japan
Benoıˆt Frisch (4), Laboratoire de Chimie Chong-Kook Kim (17), College of Phar-
Bioorganique, UMR 7514 CNRS-ULP, macy, Seoul National University, San
FacultedePharmacie,74RouteduRhin, 56-1, Shinlim-Doug, Kwanak-Gu, Seoul,
Illkirch67400,France SouthKorea
contributors to volume373 xi
Kenji Kono (27), Department of Applied Soo-JeongLim(17),CollegeofPharmacy,
Materials Science, Graduate School of Seoul National University, San 56-1,
Engineering, Osaka Prefecture Univer- Shinlim-Doug,Kwanak-Gu,Seoul,South
sity, 1-1, Gakuencho, Sakai, Osaka Korea
599-8531,Japan
Yong Liu (34), Geraldine Brush Cancer
Krystyna Konopka (31), Department of Research Institute, 2330 Clay Street,
Microbiology, University of the Pacific SanFrancisco,California94115
SchoolofDentistry,2155WebsterStreet,
PatrickMachy(7),Centred’Immunologie
SanFrancisco,California94115
de Marseille-Luminy, Campus de Lu-
LakshmiKrishnan(11),InstituteforBio- miny, Case 906, 13288 Marsielle Cedex
logical Sciences, National Research 09,France
Council of Canada, 100 Sussex Drive,
Ottawa,OntarioK1A0R6,Canada Miguel Mano (19), Department of Bio-
chemistry,FacultyofSciencesandTech-
PeterE.Jensen(8),DepartmentofPath- nology, University of Coimbra, 3000
ology and Laboratory Medicine, Emory Coimbra,Portugal
University School of Medicine, Atlanta,
Georgia30322 GaryR.Matyas(3),DepartmentofMem-
brane Biochemistry, Walter Reed Army
LawrenceB.Lachman(6),Departmentof
Institute of Research, Washington, D.C.
Bioimmunotherapy,Box422,TheUniver-
20307
sity of Texas MD Anderson Cancer
Center, 1515 Holcombe Blvd., Houston, NathalieMignet(23),UMR7001,Labor-
Texas,77030 atoiredeChimieBioorganiquetetdeBio-
Olivier Lambert (29), Institut Curie, technologie Moleculaire et Cellulaire,
Ecole National Superieure de Chimie de
Section de Recherche, UMR-CNRS 168
Paris, 13 Quai Jules Guesde, BP 14,
et LRC-CEA 8, 11 rue Pierre et Marie
94403VitrysurSiene,France
Curie,75231Paris,France
Peter Laverman (15), Department of Nu- Janos Milosevits (10), Department of
clearMedicine(565),UniversityMedical Membrane Biochemistry, Walter Reed
Center Nijmegen, P.O. Box 9101, 6500 ArmyInstituteofResearch,Washington,
HBNijmegen,TheNetherlands D.C.20307
Paul J. Lee (14), Vitreoretinal Surgical AlexeyMoskovtsev(28),VNOrekhovich
Fellow, Tulane University Medical Institute of Biomedical Chemistry, Rus-
Center, 1430 Tulane Avenue, New sian Academy of Medical Sciences, 10,
Orleans,Louisiana70112 Pogodinska ya Street, 119832 Moscow,
Russia
Lee Leserman (7), Centre d’Immunologie
de Marseille-Luminy, Campus de Lu- Jean M. Muderhwa (3), Department of
miny, Case 906, 13288 Marsielle Cedex Membrane Biochemistry, Walter Reed
09,France ArmyInstituteofResearch,Washington,
D.C.20307
SongLi(21),CenterforPharmacogenetics,
SchoolofPharmacy,UniversityofPitts- Makiya Nshikawa (25), Graduate School
burgh, 633 Salk Hall, Pittsburgh, Penn- of Pharmaceutical Sciences, Kyoto Uni-
sylvania15213 versity,Sakyo-ku,Kyoto606-850,Japan
xii contributors to volume 373
VolkerOberle(18),DepartmentofMem- Mangala Rao (2), Department of Mem-
brane Cell Biology, University of Gron- brane Biochemistry, Walter Reed Army
ingen, Antonius Deusinglaan 1, 9713 AV Institute of Research, Washington, D.C.
Groningen,TheNetherlands 20307
Hayat O¨nkyu¨ksel (13), Departments of Jean-Louis Rigaud (29), Institut Curie,
Pharmaceutics and Pharmacodynamics, Section de Recherche, UMR-CNRS 168
University of Illinois at Chicago, 833 et LRC-CEA 8, 11 rue Pierre et Marie
WoodStreet,Chicago,Illinois60612 Curie,75231Paris,France
Bu¨lentO¨zpolat(6),DepartmentofBioim- Audrey Roth (4), Laboratoire de Chimie
munotherapy,Box422,TheUniversityof
Bioorganique, UMR 7514 CNRS-ULP,
TexasMDAndersonCancerCenter,1515 FacultedePharmacie,74RouteduRhin,
HolcombeBlvd.,Houston,Texas,77030
Illkirch67400,France
WilliamM.Pardridge(32),Universityof
StephenW.Rothwell(2),Departmentof
California-LosAngeles,WarrenHall,13-
Membrane Biochemistry, Walter Reed
164, 900 Veteran Avenue, Los Angeles,
ArmyInstituteofResearch,Washington,
California90024
D.C.20307
GirishchandraB.Patel(11),Institutefor
Biological Sciences, National Research Israel Rubinstein (13), Departments of
Council of Canada, 100 Sussex Drive, Pharmaceutics and Pharmacodynamics,
University of Illinois at Chicago, 833
Ottawa,OntarioK1A0R6,Canada
WoodStreet,Chicago,Illinois60612
Ve´roniquePector(20),CenterforStruc-
tural Biology and Bioinformatics, CP Jean-MarieRuysschaert(20),Centerfor
206/2, Campus Plaine-ULB, Blv du Structural Biology and Bioinformatics,
Triomphe,1050Brussels,Belgium CP 206/2, Campus Plaine-ULB, Blv du
Triomphe,1050Brussels,Belgium
MariaC.PedrosoDeLima (19,24),De-
partmentofBiochemistry,FacultyofSci- Sandor Savay (10), Department of Mem-
ences and Technology, University of brane Biochemistry, Walter Reed Army
Coimbra,3000Coimbra,Portugal Institute of Research, Washington, D.C.
20307
Nuno Penacho (19), Department of Bio-
chemistry,FacultyofSciencesandTech- Daniel Scherman (23), UMR 7001, La-
nology, University of Coimbra, 300
boratoire de Chimie Bioorganiquet et de
Coimbra,Portugal
Biotechnologie Moleculaire et Cellulaire,
Gholam A. Peyman (14), Ophthalmology Ecole National Superieure de Chimie de
Department (SL 69), Tulane University Paris, 13 Quai Jules Guesde, BP 14,
Medical Center, 1430 Tulane Avenue, 94403VitrysurSiene,France
NewOrleans,Louisiana70112
Francis Schuber (4), Laboratoire de Chi-
PedroPires(24),CenterforNeuroscience mie Bioorganique, UMR 7514 CNRS-
andCellBiology,UniversityofCoimbra, ULP, Faculte de Pharmacie, 74 Route
3000Coimbra,Portugal duRhin,Illkirch67400,France
OlgaPodobed(28),VNOrekhovichInsti- KarineSerre(7),Centred’Immunologiede
tute of Biomedical Chemistry, Russian Marseille-Luminy, Campus de Luminy,
AcademyofMedicalSciences,10,Pogo- Case 906, 13288 Marsielle Cedex 09,
dinskayaStreet,119832Moscow,Russia France
contributors to volume373 xiii
Se´rgioSimo˜es(19,24),DepartmentofBio- Larry E. Westerman (8), VGS/DVRD/
chemistry,FacultyofSciencesandTech- NCID, Centers for Disease Control and
nology, University of Coimbra, 3000 Prevention,Atlanta,Georgia30322
Coimbra,Portugal
Barbara Wetzer (23), UMR 7001,
G. Dennis Sprott (11), Institute for Bio- Laboratoire de Chimie Bioorganiquet
logical Sciences, National Research et de Biotechnologie Moleculaire et
Council of Canada, 100 Sussex Drive, Cellulaire, Ecole National Superieure de
Ottawa,OntarioK1A0R6,Canada Chimie de Paris, 13 Quai Jules
Gert Storm (15), Department of Pharma- Guesde, BP 14, 94403 Vitry sur Siene,
France
ceutics,UtrechtInstituteforPharmaceut-
icalSciences(UIPS),UtrechtUniversity, JanWilschut(5),DepartmentofMedical
TheNetherlands
Microbiology, Molecular Virology
Janos Szebeni (10), Department of Mem- Section, University of Groningen, 9713
brane Biochemistry, Walter Reed Army AVGroningen,TheNetherlands
Institute of Research, Washington, D.C.
Seiji Yamamoto (30), Division of Gene
20307
Therapy Science, Graduate School of
Toru Takagishi (27), Department of Ap- Medicine, Osaka University, 2-2 Yama-
pliedMaterialsScience,GraduateSchool da-oka, Suita City, Osaka 565-0871,
ofEngineering,OsakaPrefectureUniver- Japan
sity, 1-1, Gakuencho, Sakai, Osaka
599-8531,Japan Fumiyoshi Yamashita (25), Graduate
School of Pharmaceutical Sciences,
Marc Thiry (20), Laboratory of Cell and
Kyoto University, Sakyo-ku, Kyoto
Tissue Biology, University of Liege, Rue
606-850,Japan
dePitteurs,Liege,Belgium
MichelVandenbranden(20),Centerfor Jing-Shi Zhang (21), Center for Pharma-
cogenetics, School of Pharmacy, Univer-
Structural Biology and Bioinformatics,
sity of Pittsburgh, 633 Salk Hall,
CP 206/2, Campus Plaine-ULB, Blv du
Pittsburgh,Pennsylvania15213
Triomphe,1050Brussels,Belgium
EstherVanKesteren-Hendrikx(1),De- RenatZhdanov(28),VNOrekhovichIn-
partment of Cell and Immunology, stitute of Biomedical Chemistry, Russian
Faculty of Medicine, Vrije Universiteit AcademyofMedicalSciences,10,Pogo-
MedicalCenter,VandeBoechhorststraat dinskayaStreet,119832Moscow,Russia
7,1081BTAmsterdam,TheNetherlands
NicoVanRooijen(1),DepartmentofCell
and Immunology, Faculty of Medicine,
Vrije Universiteit Medical Center, Van
de Boechhorststraat 7, 1081 BT Amster-
dam,TheNetherlands
[1] depletion of macrophagesbyliposomes 3
[1] ‘‘In Vivo’’ Depletion of Macrophages
by Liposome-Mediated ‘‘Suicide’’
By Nico van Rooijen and Esther van Kesteren-Hendrikx
Introduction
Macrophages are multifunctional cells. They play a key role in natural
and acquired host defense reactions, in homeostasis, and in the regulation
ofnumerousbiologicalprocesses.Themaintoolstheyusetoachievethese
goals are phagocytosis followed by intracellular digestion, and production
andreleaseofsolublemediatorssuchascytokines,chemokines,andnitric
oxide(NO).Macrophagescanbefoundasresidentcellsinallorgansofthe
body, andthey canbe recruited tositesof inflammation.Their immediate
precursorsaremonocytes,whicharereleasedinthebloodcirculationfrom
thebone marrow. Aftersome time,monocytesleave the circulation, cross
thebarrier formed by thewalls ofblood vessels, and enter into one ofthe
organs,wheretheirfinaldifferentiationintomaturemacrophageswilltake
place.
Depletion of macrophages followed by functional studies in such
macrophage-depleted animals forms a generally accepted approach to es-
tablishtheirroleinanyparticularbiomedicalphenomenon.Earlymethods
for depletion of macrophages were based on the administration of silica,
carrageenan, or by various other treatments. However, incompleteness of
depletion, and even stimulation of macrophages, as well as unwanted
effects on nonphagocytic cells, were obvious disadvantages.1
For that reason, we have developed a more sophisticated approach,
based on the liposome-mediated intracellular delivery of the bisphospho-
nate clodronate.2,3 In this approach, liposomes are used as a Trojan horse
togetthesmallclodronatemoleculesintothemacrophage.Onceingested
bymacrophages,thephospholipidbilayersofliposomesaredisruptedunder
theinfluenceoflysosomalphospholipases.Thestronglyhydrophilicclodro-
nate molecules intracellularly released in this way do not escape from
the cell, because they will not easily cross its cell membranes. As a result,
the intracellular clodronate concentration increases as more liposomes
areingestedanddigested.At acertain clodronateconcentration,irrevers-
ibledamagecausesthemacrophagetobekilledbyapoptosis.4,5Clodronate
1N.vanRooijenandA.Sanders,J. Leuk.Biol.62,702(1997).
2N.vanRooijenandR.vanNieuwmegen,CellTiss.Res.238,355(1984).
3N.vanRooijenandA.Sanders,J. Immunol.Meth.174,83(1994).
Copyright2003,ElsevierInc.
Allrightsreserved.
METHODSINENZYMOLOGY,VOL.373 0076-6879/03$35.00
4 liposomes in immunology [1]
molecules released in the circulation from dead macrophages will not enter
cells again, because they are not able to cross cell membranes. Moreover,
free clodronate molecules show an extremely short half-life in circulation
and body fluids. They are removed by the renal system. The combination
of low toxicity and short half-life of clodronate makes this drug the
best choice for the liposome-mediated elimination of macrophages
‘‘in vivo’’. Clodronate in its free form is used widely as a drug for the treat-
ment of malignant hypercalcemia6 and painful bone metastasis caused by
hormone-refractory prostate cancer,7 emphasizing its nontoxic nature.
Clodronate Liposomes in Research
Clodronate Liposomes as a Tool to Investigate Macrophage
Activities In Vivo
With the liposome-mediated macrophage ‘‘suicide’’ approach, func-
tional aspects of macrophages have been established in hundreds of studies
up to now. Many of the resulting publications are listed in the website
http://www.ClodronateLiposomes.com.
Althoughliposomescannotcrossvascularbarrierssuchasthewallsof
capillaries, their meeting with particular macrophage populations can be
achieved by choosing the right administration routes. Among the macro-
phages that might become useful targets for manipulation by liposomes
are Kupffer cells in the liver and splenic macrophages (to be reached by
way of intravenous injection),8 alveolar macrophages in the lungs (to be
reached by way of intratracheal instillation or intranasal administration),9
phagocytic synovial lining cells (by means of intra-articular injection in
the synovial cavity),10 peritoneal macrophages (by means of intraperito-
nealinjection),11macrophagesinthetestis(bymeansoflocalinjection),12
4N.vanRooijen,A.Sanders,andT.vandenBerg,J. Immunol.Meth.193,93(1996).
5M. Naito, H. Nagai, S. Kawanao, H. Umezu, H. Zhu, H. Moriyama, T. Yamamoto,
H.Takatsuka,andY.Takkei,J. Leuk.Biol.60,337(1996).
6A.List,Arch.Intern.Med.151,471(1991).
7A.Heidenreich,R.Hofmann,andU.H.Engelmann,J. Urol.165,136(2001).
8N. van Rooijen, N. Kors, M. van den Ende, and C. D. Dijkstra, Cell Tiss. Res. 260, 215
(1990).
9T.Thepen,N.vanRooijen,andG.Kraal,J. Exp.Med.170,499(1989).
10P.L.E.M.vanLent,A.E.M.Holthuyzen,L.A.M.vandenBersselaar,N.vanRooijen,
L.A.B.Joosten,F.A.J.vandeLoo,L.B.A.vandePutte,andW.B.vandeBerg.Arthritis
Rheum.39,1545(1996).
11J.Biewenga,B.vandeEnde,L.F.G.Krist,A.Borst,M.Chufron,andN.vanRooijen,Cell
Tiss.Res.280,189(1995).
12A.Bergh,J.E.Damber,andN.vanRooijen,J. Endocrinol.136,407(1993).