Institute of Immunology

Annual report 2006

Contents

Institute of Immunology Rikshospitalet-Radiumhospitalet Medical Center

and University of Oslo Oslo, Norway

Annual report 2006 Contents INTRODUCTION…………………………………………………………………………………………………………….... 3 FACULTY MEMBERS AND MANAGEMENT 2006……………………………………………………………………… 5

5 HONOURS IN 2006……………………………………………………………………………………………………. THIMMI ORGANIZED THE 20 EFI CONFERENCE IN OSLO, 8-11 JUNE.………………….………………………… 6

RESEARCH GROUPS……………………………………………………………………………………………….……… 8 COMPLEMENT RESEARCH GROUP……………………………………………………………………………….

10 IMMUNOGENETICS OF AUTOIMMUNE DISEASES………………………………………………………………

13 IMMUNOLOGY OF MULTIPLE SCLEROSIS……………………………………………………………………….

14 NK CELL RESEARCH GROUP………………………………………………………………………………………

16 THE CELL THERAPY RESEARCH GROUP………………………………………………………………………..

18 THE FUNCTIONAL IMMUNOGENETICS GROUP…………………………………………………………………

20 THE PROTEOMICS GROUP………………………………………………………………………………………….

22 TUMOR IMMUNOLOGY, AUTOIMMUNITY AND VACCINE DEVELOPMENT…………………………………

SERVICE SECTIONS………………………………………………………………………………………………………… 25 SECTION OF MEDICAL IMMUNOLOGY (MI)……………………………………………………………………….

26 SECTION OF TRANSPLANTATION IMMUNOLOGY (TI)………………………………………………………….

TEACHING…………………………………………………………………………………………………………………….. 28 RESULTS OF OUR RESEARCH 2006……………………………………………………………………………………..

29 DOCTORAL THESES……………………………………………….………………………………………………….

29 THESIS FOR MASTER OF SCIENCE…………………………………………………..……………………………

29 BOOKS AND BOOK CHAPTERS………………………………….………………………………………………….

29 PUBLICATIONS …………………………………………………….………………………………………………….

33 ACKNOWLEDGEMENT………………………………………………………………………………………………..

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Introduction

Introduction INSTITUTE OF IMMUNOLOGY (IMMI) The Institute of Immunology (IMMI) was established in 1998, as a merger between the Institute of Rheumatology and Immunology (IGRI) and the Institute of Transplantation Immunology (ITI). It is a combined institute at the Rikshospitalet-Radiumhospitalet Medical Center and the University of Oslo, with a joint and fully integrated administration. The aim of the institute is to be a major service institute and centre of competence in clinical immunology in Norway, and to be a research institute contributing to the advancement of basic and clinical immunology on an international level. The clinical service functions cover major parts of medical immunology, and were for the most part of 2006 organized into two sections, Medical Immunology (MI) and Transplantation Immunology (TI), as well as an advanced Ex vivo Laboratory of Cell Therapy. The research functions are carried out in several research groups, with a close collaboration between them. The institute also has extensive teaching functions in basic and clinical immunology. This includes medical students, students in other fields such as biology, as well as at postgraduate levels. The present staff is comprised of approximately 140 persons, including medical doctors, scientists, research fellows, technologists and other personnel. This annual report for 2006 is a follow up of our previous annual reports. Our web site, www.med.uio.no/rh/immi, contains more details about the institute and its current activities. ERIK THORSBY RESIGNED AS HEAD OF IMMI Erik Thorsby resigned as head of the Institute of Immunology 1st September 2006. A provisional status of the institute was given by Erik in his last full year as head, see the annual report from 2005. The university part of the institute has been without a formal leader since Erik’s resignation, John Torgils Vaage has acted as head of the hospital part for most of the year (during Erik’s sabbatical in the first half of the year and after September 1).

From the Jubilee dinner: Sir Walter Bodmer, Erik, Frans Claas, Dominique Charron and Peter Parham

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Introduction

Erik established the HLA field in Norway. He also established the Institute of Transplantation Immunology (ITI) in 1970, and became its first head. When the Institute of Transplantation Immunology merged with the Institute of Rheumatology and Immunology (IGRI) in 1998, Erik became the first head of this new Institute of Immunology, a position he kept till 1st September 2006. From the same date Erik has a part time position as professor and head physician at the institute, where he together with Benedicte A. Lie are group leader of the Research group of Immunogenetics of Autoimmune diseases. In connection with Erik’s resignation as the head of the institute, the institute organized a symposium on 15th September entitled: “40 years with HLA”, in the large auditorium at Rikshospitalet, chaired by Frode Vartdal and John Torgils Vaage. National and international national acknowledged experts in the field, who have become close friends of Erik, lectured at the symposium: Peter Parham (US), Frans Claas (NL), Dag Undlien (N), Dominique Charron (F), Ludvig Sollid (N), Gustav Gaudernack (N) and Sir Walter Bodmer (UK). Later the same day the institute arranged a Jubilee dinner at Det Norske Medicinske Selskap. BJARTE G. SOLHEIM RESIGNED AS HEAD OF THE SECTION FOR MEDICAL IMMUNOLOGY (MI)

stBjarte G. Solheim resigned 1 August 2006 and was succeeded by Johanna Olweus. Bjarte has been at Rikshospitalet since 1972, when he started as chief resident at the Institute of Transplantation Immunology (ITI). From 1989 to 1995 he was in charge of transfusion medicine at the Rikshospitalet. In 1996 he returned to ITI (later IMMI) and established the Section for Advanced Transfusion Medicine, which became GMP (Good Manufacture Production) certified for cytapheresis of hematopietic stem cells, lymphocytes, dendritic cells, granulocytes and leukemic cells. After the fusion of Rikshospitalet and Radiumhospitalet these activities were moved to Montebello, and Bjarte took over as the head of the institute’s Section for Medical Immunology in 2005. In connection with Bjarte’s resignation, the institute and the Norwegian Association for Immunology and Transfusion Medicine (NFIT) organized a seminar on November 21-22 entitled: “Current Topics in Transfusion Medicine” at Rikshospitalet. National and international experts in the field, who have become close friends of Bjarte, presented lectures at the symposium: Peter Hellstern (D), Ulrich Sachs (D), Martin L. Olsson (SE), Amy Tsai (US), Tor-Einar Svae (AU), Jerard Segatchian (UK), Dick van Rhenen (NL), Anne Husebekk (N) and Torunn Oveland Apelseth (N). Articles based on the lectures are published in Transfusion and Apheresis Science (November 2007). On the evening of November 21 the institute sponsored a Jubilee dinner at Bjarte’s residence.

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Faculty members and management 2006

Faculty members and management 2006 Thorsby, Erik, MD Head of the institute, (to August), Professor Vaage, John Torgils, MD Acting head of the institute (hospital part: Jan-June, Sept-Dec) Head, Section of Transplantation Immunology (to June) Berger, Lisbeth Office manager Sundsten, Brit Chief technologist Bogen, Bjarne, MD Professor, Head physician Brinchmann, Jan E., MD Senior scientist, Head. Ex Vivo Laboratory of Cell Therapy Egeland, Torstein, MD Head, Section of Transplantation Immunology (from July) Fleckenstein, Burkhard, PhD Senior scientist Lea, Tor, PhD Senior scientist, Professor Leivestad, Torbjørn, MD Head physician Lie, Benedicte A., PhD Senior scientist Mollnes, Tom Eirik, MD Professor, Head physician Olweus, Johanna, MD Head, Section of Medical Immunology (from August) Solheim, Bjarte G., MD Head, Section of Medical Immunology (to July), Professor Sollid, Ludvig M., MD Professor, Head physician *Vartdal, Frode, MD Head physician (part time) *Frode Vartdal served as Head of the Clinic of Laboratory Medicine in the first half and as Head of the Faculty Division Rikshospitalet (University of Oslo) in the second half of 2006. HONOURS IN 2006 The groups of Bjarne Bogen and Ludvig M. Sollid (which included Burkhard Fleckenstein) have been selected by the Research Council of Norway to become a Centre of Excellence (Centre of Immune Regulation (CIR) led by Ludvig M. Sollid). Ludvig M. Sollid received the Research Council of Norway's Møbius Prize for Outstanding Research. Tom Hemming Karlsen and coworkers received from Helse Sør the Prize for the best article on basic medical sciences published in the second half of 2006, published in Gastroenterology. 2006; 131: 781-7. Stig Tollefsen, Helene Arentz-Hansen and coworkers received from Helse Sør the Prize for the best article on clinical medical sciences published in the second half of 2006, published in Journal of Clinical Investigation 2006; 116: 2226-36. Trygve Holmøy and coworkers received from the Norwegian Society for Immunology Prize for the best article in immunology published in 2004-2005, published in European Journal of Immunology. 2005; 35: 1786-94. Trygve Holmøy received The Monrad Krohn's Prize for the Advancement of Neurological Research 2006.

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20th EFI Conference

IMMI organized the 20th EFI Conference, Oslo 8-11 June The European Federation for Immunogenetics (EFI), is an organization of scientists working in the field of immunogenetics and histocompatibility testing in Europe. Its 20th annual conference was held in Oslo 8.-11. June, organized by IMMI. It was coined “From gene structure to gene function” to emphasize a focus on functional immunogenetics. The Local Organizing Committee consisted of Erik Thorsby (chairman), Irene Andersen, Lisbeth Berger, Torstein Egeland, Benedicte A. Lie, Johanna Olweus, Anne Kari Tveter and John Torgils Vaage. 650 participants from Europe and also many other countries, attended the conference. In addition there were 41 accompanying persons, which resulted in a total attendance of 691 people. To our knowledge this is the largest number of participants to attend an EFI conference. The opening lecture, the Ceppellini Award lecture, was given by Rolf Zinkernagel, the Nobel Prize laureate in Medicine in 1996. James Traherne from UK gave the Julia Bodmer Award lecture. There were five plenary sessions, covering various important topics in immunogenetics such as an update on recent molecular developments, the role in innate and adaptive immunity, disease predisposing genes, histocompatibility testing and matching and the future of immunogenetics. The plenary talks were given by highly acknowledged international experts in their respective fields.

Left:Zinkernagel Under: Traherne

370 abstracts were received, a new record for any EFI conference. The accepted abstracts were presented in oral and poster sessions. Further five teaching sessions for students and technologists in the field were held.

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20th EFI Conference

The social programme included a reception given by the Mayor of Oslo in the City Hall, a Conference Dinner at The Norwegian Maritime Museum with visits also to the Fram and Kon-Tiki museums. In addition one of the sponsors, One Lambda, gave a “Wild Night” at one of Oslo’s Dancing Restaurants. The feed back from the participants was very positive. The extremely nice weather with clear skies throughout the whole conference contributed to its success. The accompanying pictures (except the picture of the congress poster) were provided by EFI Newsletter.

Conference Dinner at The Norwegian Reception at Oslo City Hall Maritime Museum

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Research groups

COMPLEMENT RESEARCH GROUP GROUP LEADER: TOM EIRIK MOLLNES, MD GROUP DESCRIPTION Complement is part of the innate immune system protecting the host against invading micro-organisms. Regulatory control mechanisms normally prevent the system from extensive and systemic activation, thereby protecting the host from self damage. Under various disease conditions complement is improperly activated, either locally leading to tissue damage or systemically with risk of serious homeostatic disturbances. A primary research goal for the Complement Research Group is to elucidate the role of complement as a primary inducer of the inflammatory reaction and thereby form a basis for a future therapeutic approach in complement-mediated disease processes. For this purpose we have developed novel assays for detection and quantification of complement activation products based on monoclonal antibodies to activation dependent epitopes on a number of complement components; the most important one being the assay for TCC (the terminal SC5b-9 complement complex). These assays are used to detect complement activation experimentally and clinically and to evaluate the effect of various complement inhibitors in experimental models. In a novel in vitro human whole blood model where all potential inflammatory mediators are able to interact mutually, we are currently studying the effect of complement inhibition on a number of arms of the inflammatory network. Current clinical and experimental animal projects focus on allo- and xenotransplant rejection, pulmonary and systemic inflammation in the meconium aspiration syndrome of the newborn, infection and septicaemia, ischemia-reperfusion injury and stroke, biocompatibility of extracorporeal circulation devices, and rheumatic tissue damage. RESEARCH SUMMARY 2006 Seven international original articles and two reviews were published from the group in 2006. The main research focus has been on the role of complement in human disease. GROUP MEMBERS 2006 Tom Eirik Mollnes (Group leader; Professor) Anne Pharo (Chief laboratory technician) Julie Lindstad (laboratory technician) Elena D. Pedersen (PhD student) Marit Sæthre (PhD student) Bodil Salvesen (PhD student) Ebbe Thorgersen (PhD student) Bernt Chr. Hellerud (PhD student) Albert Castellheim (Postdoc) Morten Harboe (Professor Emeritus) COLLABORATIONS 2006 *National Prof. Helge Scott, Institute of Pathology, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Dr. Guttorm Haraldsen, Institute of Pathology, Rikshospitaley University Hospital, Oslo, Norway Prof. Ola Didrik Saugstad, Dept. of Pediatric Research, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Dr. Arnt E. Fiane, Department of Thoracic and Cardiovascular Surgery, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Prof. Jan L. Svennevig, Department of Thoracic and Cardiovascular Surgery, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Prof. Odd Geiran, Department of Thoracic and Cardiovascular Surgery, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway

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Research groups

Dr. Svein S. Simonsen, Department of Cardiology, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Dr. Lars Gullestad, Department of Cardiology, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Prof. Stig S. Frøland, Section of Clinical Immunology and Infectious Diseases, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Dr. Pål Aukrust, Section of Clinical Immunology and Infectious Diseases, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Hilde Fure, Grethe Bergseth and Dorte Christensen, Nordland Hospital, Bodø, Norway Dr. Knut Tore Lappegård, Nordland Hospital, Bodø, Norway Prof. Erik Waage Nielsen, Dept. of Anesthesiology, Nordland Hospital, Bodø, Norway Prof. Ole-Lars Brekke, Dept. of Medical Biochemistry, Nordland Hospital, Bodø, Norway Prof. Peter Kierulf, Ullevaal University Hospital, Oslo, Norway Prof. Petter Brandzaeg, Ullevaal University Hospital, Oslo, Norway Prof. Vibeke Videm, Trondheim University Hospital, Trondheim, Norway *International Dr. Michael Fung, Tanox. Inc, Houston, Texas, USA Dr. John D. Lambris, Philadelphia, Pennsylvania, USA Dr. Peter Garred, Copenhagen, Denmark Dr. Johan Riesenfeld, Carmeda AB, Stockholm, Sweden Dr. Tom Sprong, Nijmegen, The Netherlands Dr. C. Erik Hack, Amsterdam, The Netherlands Dr. Moh R. Daha, Leiden, The Netherlands Dr. Anja Roos, Leiden, The Netherlands Dr. Greg Stahl, Harward Medical School, Boston, USA Dr. Franco Tedesco, Trieste, Italy

Anne Pharo

Chief laboratory technician

Albert Castellheim Post Doc

Elena Pedersen

PhD. student

Morten Harboe

Professor emeritus

Tom Eirik Mollnes

Professor/group leader

Marit Sæthre PhD. student

Julie Katrine Lindstad

Laboratory technician

2006

Bodil Salvesen PhD. student

Bernt Chr. Hellerud PhD. student

Ebbe Thorgersen PhD. student

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Research groups

IMMUNOGENETICS OF AUTOIMMUNE DISEASES GROUP LEADERS: BENEDICTE A. LIE, PhD AND ERIK THORSBY, MD GROUP DESCRIPTION Autoimmune diseases are caused by a combination of genetic and environmental factors. The main interest of the group is to identify genes that are predisposing to the development of autoimmune diseases, like type 1 diabetes (T1D), multiple sclerosis, juvenile rheumatoid arthritis, primary sclerosing cholangitis (PSC), rheumatoid arthritis, inflammatory bowel disease, systemic lupus erythematosus, myasthenia gravis etc. Our work is mainly focused on the many genes in the HLA complex, which is contributing the most to the genetic risk for these diseases. Through our studies over the last 20 years, we have contributed to a better identification of the primary disease-predisposing HLA complex genes, which seem to be genes encoding some peptide-presenting HLA class II molecules. During recent years we have focused our studies on unravelling novel disease-predisposing genes in the extended HLA complex. By systematically screening the HLA complex and controlling for already known risk factors, we found evidence for several additional predisposing genes for autoimmune diseases. Our current work is focused on the identification of these “novel” predisposing genes and their possible functions. RESEARCH SUMMARY 2006 During the last year, our research has focused on refining the associations located in the HLA complex to various autoimmune diseases by performing extensive screens using hundreds of SNPs, as well as complete HLA genotyping of both HLA class I and II loci. These studies have pointed out novel and interesting genes and regions of the MHC of importance in the diseases under study. We are also actively participating in the MHC fine mapping project managed by the T1D genetic consortium (T1DGC). We are one of the data analysis teams performing statistical analyses on the large amount of genotype data (2 957 SNPs, 66 microsatellites and complete HLA typing) generated on 11 279 individuals in 2 322 type 1 diabetes families from all over the world. All teams will present their results at a meeting in Washington in August 2007. Albeit the fact that 80% of PSC patients also have inflammatory bowel disease, mainly ulcerative colitis, the HLA-DRB1 and DQB1 alleles strongly associated with PSC did not show significant association with ulcerative colitis, indicating that the ulcerative colitis seen in PSC may be a distinct disease phenotype. In addition to HLA, we have also studied the killer immunoglobulin-like receptors (KIR) genes in PSC. No disease association was detected at the KIR level, however, a strong association was found with KIR ligands encoded by three neighboring genes in the HLA complex. We have also tested genes elsewhere in the genome for possible involvement in autoimmune diseases. Polymorphisms in T cell signal transduction genes were studied in multiple sclerosis without any associations being revealed, neither for the PTPN22 1858T allele, known to predispose to a number of other autoimmune diseases like rheumatoid arthritis. Studies of the PTPN22 gene of our Norwegian rheumatoid arthritis patients showed that the association with a promoter polymorphism -1123G>C could not be distinguished from the 1858C>T association pointing out that the genetic association in this region has not been fully dissected. Interestingly, we found that the PTPN22 1858T allele is associated with increased progression rate of radiographic joint destruction in rheumatoid arthritis. Another finding of genetic association with clinical outcome was that polymorphisms in the SXR gene seem to influence disease progression in PSC. We also performed DNA typing of some serum samples from Easter Island, collected in 1971 from reputedly non-admixed native Easter Islanders. All individuals carried mtDNA types and HLA alleles previously found in Polynesia, and most men carried Y chromosome markers of Polynesian origin,

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Research groups

providing further evidence of a Polynesian origin of the population of Easter Island. A few individuals carried HLA alleles and/or Y chromosome markers of European origin. More interestingly, some individuals carried the HLA alleles A*0212 and B*3905, which are of typical Amerindian origin. The presence of these Amerindian HLA alleles could be traced back to at least 1815. Thus, our investigations strongly suggest an early Amerindian contribution to the Polynesian gene pool of Easter Island. GROUP MEMBERS 2006 Erik Thorsby (Group leader) Benedicte A. Lie (Group leader) Hanne F. Harbo (Post doc) Marte K. Viken (PhD student) Tom H. Karlsen (PhD student) Kristine Wiencke (PhD student) Morten C. Eike (PhD student) Gry B. Namløs Nordang (PhD student) Espen Melum (Medical student) Åslaug R. Lorentzen (PhD student) Siri Flåm (Research assistant) Hege Dahlen Sollid (Research assistant) Tone Aarskaug/Bente Woldseth (Research assistant) COLLABORATIONS 2006 *National Kirsten M. Boberg and Erik Schrumpf, Dep. of Medicine, Rikshospitalet-Radiumhospitalet Medical Center, Oslo Knut Dahl-Jørgensen and Geir Joner, Dep. of Paediatrics, Ullevål Univ. Hospital, Oslo Berit Dupuy, Institute of Forensic Medicine, University of Oslo, Oslo Per Ekstrøm, Department of Surgical Oncology, The Norwegian Radium Hospital, Oslo Berit Flatø and Øystein Førre, Center for Rheumatic diseases, Rikshospitalet-Radiumhospitalet Medical Center, Oslo Kathrine Frey Frøslie, Section of Biostatistics, Rikshospitalet-Radiumhospitalet Medical Center, Oslo Elisabeth Gullowsen Celius, Department of Neurology, Ullevål University Hospital, Oslo Erika Hagelberg, Department of Biology, University of Oslo, Oslo, Norway Kristian F. Hanssen, Department of Endocrinology, Aker University Hospital, Oslo Tore K. Kvien, Department of Rheumatology, Diakonhjemmet Hospital, Oslo Svein-Ivar Mellgren, Department of Neurology, University Hospital of North Norway, Troms Kjell-Morten Myhr, Department of Neurology, Haukeland University Hospital, Bergen Anne Spurkland, Institute of Anatomy, University of Oslo, Oslo Dag Undlien, Institute of Medical Genetics, Ullevål University Hospital, Oslo Kjersti Skjold Rønningen, Dep. of Population Health Sciences, National Institute of Public Health, Oslo Chantal Tallaksen, Department of Neurology, Ullevål University Hospital, Oslo Bodvar Vandvik, Department of Neurology, Ullevål University Hospital, Oslo Morten Vatn and the IBSEN study group, Egil Utsi, Karasjok Heart Center, Karasjok *International Christopher L. Bowlus, Division of Gastroenterology, Univ. of California Davis Medical Center, USA Annika Bergquist, Division of Gastroenterology and Hepatology, Karolinska Univ. Hospital, Sweden Anne Cambon-Thomsen, Inserm U 558 Faculte de Medicine, Toulouse, France Marcello Fernadez-Vina, Dept. of Laboratory Medicine, Univ. of Texas, Houston, Tx, USA The GAMES group, lead by A. Compston, Neurology Unit, Addenbrookes Hosp., Univ. of Cambridge Ingrid Kockum, Department of Molecular Medicine, Karolinska Institutet, Stockholm, Sweden. Jørn Nerup and Flemming Pociot, Steno Diabetes Center, Gentofte, Denmark The Nordic Study Group on Multiple Sclerosis Genetics, lead by Jan Hillert, Department of Neurology, Karolinska Institute, Stockhom Marita Olsson, Dep. of Mathematical Statistics, Chalmers University of Technology, Göteborg, Sweden Stefan Schreiber and Jochen Hampe, Dep. of Medicine, Christian-Albrechts University, Kiel, Germany Ji-Yao Sun, Div. of Hemat. and Bone Marrow Transpl., City of Hope National Medical Center, CA, USA

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Research groups

Eva Tolosa, University of Tübingen, Germany

The Immunogenetics group

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Research groups

IMMUNOLOGY OF MULTIPLE SCLEROSIS GROUP LEADER: FRODE VARTDAL, MD RESEARCH SUMMARY 2006 The research focus is immunology of neurological diseases, particularly multiple sclerosis (MS) and stiff person syndrome (SPS). The goal is to identify therapeutic targets, and the research is both directed against mechanisms of disease and mechanism of action of current treatment. We study lymphocytes and antibodies from the cerebrospinal fluid (CSF), which probably reflects the pathogenic process better than blood. MS: Mechanisms of disease. Whereas animal model of MS are mediated by T cells specific for myelin antigens, the target antigens in MS have not been identified. We have postulated that the immune response in MS is driven by T cells recognizing mutated fragments on the variable regions on IgG expressed and secreted by autologous B cells. To address this we have cloned T cells and B cells from the CSF of MS patients. We have recently demonstrated that T cells target several different mutated fragments on the same autologous CSF IgG molecule. These results suggest that mutated CSF IgG could mediate intrathecal IgG production and inflammation in MS. MS: Treatment. Glatiramer acetate (GA) is a synthetic random copolymer used in immunotherapy of MS. GA is supposed to induce GA reactive T cells secreting anti-inflammatory cytokines, but the mechanism has so far not been studied in T cells from the intrathecal compartment of humans. We have generated GA reactive T cells from CSF and blood of MS patients prior to and after GA treatment, and demonstrated that GA reactive T cells from CSF display pronounced and significant differences compared to GA reactive T cells from blood. SPS: SPS is a rare neurological disease associated with antibodies against the 65-kDa isoform of glutamate decarboxylase (GAD65). We have demonstrated that SPS patients synthesise oligoclonal GAD65-specific IgG in the intrathecal compartment. Moreover, we have cloned GAD65 specific T cells from blood and CSF, and mapped the target T cell epitopes on GAD65. GROUP MEMBERS Frode Vartdal, (Group leader) Trygve Holmøy (Postdoc) Anne Lise Karlsgot Hestvik (Research fellow) Gjertrud Skorstad (Reseach Fellow) COLLABORATORS 2006 Professor Bjarne Bogens group, Institute of immunology, Rikshospitalet-Radiumhospitalet Medical Center Professor Anne Spurklands group, Institute of Basic Medical Sciences, Department of Anatomy, University of Oslo Professor (em) Bodvar Vandvik, Department of Neurology, Ullevål University Hospital Head of department, Elisabeth Gullowsen Celius, Department of Neurology, Ullevål University Hospital Professor David Price, 3Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, UK

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Research groups

NK CELL RESEARCH GROUP GROUP LEADER: JOHN T. VAAGE, MD GROUP DESCRIPTION The NK cell research group (NKRG) studies the biology and recognition mechanisms of Natural Killer (NK) cells. Focus has been on an experimental animal model in rats, but studies of human NK receptors (KIRs) are in progress. There has been a longstanding collaboration with Prof. Bent Rolstad at the Department of Anatomy, Institute of Basic Medical Sciences. The two laboratories are at different locations within the Rikshospitalet/University campus, but are functioning as a combined group with nationally and EU funded projects. RESEARCH SUMMARY 2006 NK cells express several families of receptors for MHC and non-MHC-encoded target cell antigens. These receptors belong to two structurally distinct superfamilies, the C-type lectin-like receptors including the killer cell lectin-like receptors (KLR) and the Ig-related receptors which include the killer cell immunoglobulin-like receptors (KIR). Most receptor families are comprised of both inhibitory and activating members, which may exhibit an overlapping specificity for ligand. We have focused on the Ly49 multigene family of rat KLR receptors, which can be viewed as functional homologues of the KIR receptor family in humans. Ly49 receptors are encoded in the rat NK gene complex, which contains more than 30 Ly49 genes. They can bind polymorphic MHC class I molecules and mediate rejection of foreign stem cell grafts. In the past year, we completed a study of four structurally related receptors, Ly49s3, -i3, -s4, -i4, all being very similar in the extracellular ligand-binding region despite having opposing effects on NK cell function. While the two inhibitory Ly49 receptors (Ly49i3/-i4) were expressed in all rat strains studied, their activating counterparts (Ly49s3/-s4) were present in only a limited panel of high NK alloresponder strains. In a separate study, it was demonstrated that Ly49 expression is not uniform among NK cells, but rather focused to a subset of highly alloreactive NK cells. The complementary NK cell subset, which expresses very little Ly49 receptors and is not alloreactive, was identified by its expression of a novel NK cell receptor (NKR-P1C). It appeared earlier in ontogeny and was predominant in certain tissues, notably in lymph nodes from the gut. These observations again point to a central role for activating Ly49 receptors in triggering NK cell alloreactivity. In ongoing studies we use a heterologous reporter system for ligand identification for rodent and human receptors. This system uses T cell lines stably transfected with GFP or the LacZ enzyme under control of the NFAT1 transcription factor, which acquire green fluorescence, or produce LacZ upon activation of tyrosine kinase coupled receptors. Receptor biology is also being pursued in vivo, in rat models for allogeneic stem cell transplantation and acute myelogeneic leukemia (AML), and in an infection model using the intracellular bacterium Listeria monocytogenes. These studies will hopefully improve our understanding of the roles played by the various activating and inhibitory NK receptors in the immune defense against certain infections and cancerous cells, and in experimental and clinical allotransplantation. GROUP MEMBERS 2006 John T. Vaage (Group leader) Lise Kveberg (Post doc) Ke-Zheng Dai (Senior research assistant) Christian Naper (Resident/ senior scientist) Michael Daws (Senior scientist) Egil Veland (Medical student) Associated group members: Bent Rolstad (Prof., Inst. Basic Med. Sciences) Bente Kahrs Omdal (Research assistant) Marit Inngjerdingen (Post doc) Hamid Shegarfi (PhD student) Janne M Nestvold (PhD student) Severin Zinöcker (PhD student)

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Research groups

COLLABORATIONS 2006 *National E. Dissen, S. Fossum, Department of Anatomy, Institute of Basic Medical Sciences, University of Oslo H. Benestad, Department of Physiology, Institute of Basic Medical Sciences, University of Oslo G. Kvalheim, Department of Medical Oncology and Radiotherapy, The University Hospital of Rikshospitalet-Radiumhospitalet Oslo *International J.C. Ryan, VA Medical Center, Univ. of California San Francisco (UCSF), USA E. Joly, Equipe de Neuro Immuno Génétique Moléculaire, Toulouse, France L. Walter, Division of Primate Genetics, German Primate Centre, Göttingen, Germany R. Dressel, Division of Immunogenetics, Univ. of Göttingen, Germany H Hedrich, Medizinische Hochschule Hannover, Germany

Some members of the combined research group (including associated members).

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Research groups

THE CELL THERAPY RESEARCH GROUP GROUP LEADER: JAN E. BRINCHMANN, MD GROUP DESCRIPTION This group was established as a fusion of Jan E. Brinchmann’s, Torstein Egeland’s and Tor Lea’s research groups. Johanna Olweus and her research associates later joined the group. The aim of the Cell Therapy Research Group is to perform basic and translational research with relevance to cell therapy. As a result of a recent re-organization, the group is now split into two thematic areas: stem cells and T cell/dendritic cell interactions RESEARCH SUMMARY 2006 The focus of the stem cell research has been on characterization of differentiation pathways of mesenchymal (MSC) and hematopoietic (HSC) stem cells from bone marrow, and stromal stem cells from adipose tissue (AT-MSC). We have established a robust cell culture system for MSC, also with autologous serum in stead of fetal bovine serum, and have characterized these cells with respect to phenotype, mRNA and protein expression profile. We have isolated uncultured AT-MSC and performed extensive charcterization of these cells and and their cultured offspring. We have established several 3-dimensional culture systems for these stromal stem cells. We are also studying the effect of inflammatory mediators on MSC, particularly how inflammation-induced intracellular signals modulate the activity of the Wnt pathway and other signaling pathways known to be involved in fate determination in stem cells. Several lines of evidence suggest that adult HSC can differentiate into neural cells, albeit at relatively low frequency. We have recently shown that adult bone marrow-derived CD34+ HSC implanted into the regenerating spinal cord of the chicken embryo produce morphologically and biochemically differentiated, synaptically integrated human neurons, including motorneurons, at substantially higher efficiency than previously obtained in other systems. This study is now focusing on which subpopulation of CD34+ HSC has the greatest potentiality for neuronal differentiation. At a more clinical level, the group has been involved in a project where bone marrow derived stem cells were used to try to regenerate myocardium following acute myocardial infarction, and in another project to try to achieve re-vascularization of critically ischemic legs. As a part of efforts to characterize cellular signalling pathways, we have developed a new proteomic platform for measuring protein phosphorylation and associations between proteins. A multiplex particle suspension array captures hundreds of different targets in a single sample. The amount of each analyte and post-translational modifications are studied by sandwich antibodies and phosphoepitope-specific antibodies. We have also established techniques for efficient purification and isolation of phosphopeptides prior to analysis by MALDI-TOF/TOF mass spectrometry.These methodological developments will be used to study activation processes in DCs, T cells and stem cells. The T-cell/dendritic cell (DC) research has focused on ways to optimalize immunotherapy of leukemias. The requirements for generating “naturally occurring” DC from CD34+ progenitors are studied. We have found dendritic cells are generated from a defined subset of CD34+ cells that expresses the transcription factor ICSBP. These progenitors are deficient in a subset of patients with chronic myeloid leukaemia. We are currently determining what cytokines control the generation of ICSBP+ progenitors. In two separate projects, we are establishing protocols for DC-mediated stimulation of T cells with antigens from whole leukemia cells on one hand or cytomegalovirus on the other. In another project the aim is to remove the cells responsible for graft versus host disease from allogeneic bone marrow, whereas cells responsible for anti-leukemic effects remain. We have also established a protocol for suicide gene transduction of allogeneic T cells to be used for donor lymphocyte infusion of patients with leukemia, and characterized the transfused cells at the clonal level. The regulatory interplay between human MSCs and T lymphocytes have been studied in detail. hMSCs constitutively produce prostaglandin E2 (PGE2) in amounts sufficient to inhibit T-cell proliferation and differentiation. Moreover, early proinflammatory cytokines released from activated T

16

Research groups

helper cells potently induce the expression of cyclooxygenase 2 (COX2) in MSCs leading to increased PGE2-production and enhancement of the immunosuppressive properties of the MSCs. The molecular details of this interaction are being further scrutinized. Another research activity within the group has concentrated on the molecular characterization of T-cell specific adapter protein (TSAd). These studies have provided new insight in the regulatory function of the TSAd protein, particularly its effect on the protein tyrosine kinase p56lck in proximal T-cell activation. Associated with the Cell Therapy Research Group, Professor Bjarte G. Solheim is performing studies on transplantation immunology and transfusion medicine. This research is focused on hematopoietic chimerism after blood transfusion or solid organ transplantation, and preservation and optimal use of blood components GROUP MEMBERS 2006 Jan E. Brinchmann (Group Leader) Torstein Egeland (Head, Section of

Transplantation Immunology) Tor Lea (Senior scientist) Johanna Olweus (Head, Section of Medical

Immunology) Fridtjof Lund-Johansen (Post doc) Katrine Frønsdal (Post Doc) Krisztina Szöke (Post doc) Xiaolin Wang (Post doc) Weiwei Wu (PhD student) Olafur E. Sigurjonsson (PhD student) Espen Kvale (PhD Student) Yngvar Fløysand, (PhD student) Charlotte Kleiveland (PhD student) Ida Aagård Hedfors (PhD Student) Aboulghassem Shahdadfar (PhD Student) Eli Taraldsrud (MD, Junior consultant) Rune Bruhn Jakobsen (Medical student) Iris Villa (Medical student) Erlend Strønen (Medical student) Janke B. Eriksen (Lab technician) Shivali Duggal (PhD student) Tommy Karlsen (Master student) Karen J. Beckstrøm (Lab. technician) Kristin Marshall (Lab technician) Ellen Karlstrøm (Lab. technician) Ingerid W. Abrahamsen (PhD. student) ASSOCIATED PERSONNEL Bjarte G. Solheim, MD, Professor Øystein Flesland, MD, Ph. D. Student COLLABORATIONS Prof. Per Brandtzæg, Institute of Pathology, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway Prof. Philippe Collas, Institute of Medical Biochemistry, University of Oslo Prof. Robert Nerem, Georgia Institute of Technology, Atlanta, USA Prof. George Daley, Harvard Medical School, Boston, USA Prof. Joel Glover, PhD; Institute of Physiology, University of Oslo, Oslo, Norway Prof. Lars Engebretsen, Dept. of Orthopedics, Ullevål University Hospital, Oslo, Norway Dr. Geir Tjønnfjord, Senior Consultant in Hematology; Section for Blood Diseases, RH, Oslo Dr. Aksel Foss, Senior Consultant in Surgery, Section for Transplantation Surgery, RH, Oslo Prof. Kolbjørn Forfang, Section for Cardiology, RH, Oslo, Norway Prof. Arnfinn Ilebekk, Institute of Experimental Medical Research, Ullevål University Hospital Prof. Anne Spurkland, Inst. of Anatomy, University of Oslo, Norway Prof. Tomas Mustelin, The Burnham Institute, La Jolla, CA, USA Dr. Burkhard Fleckenstein, Institute of Immunology, Rikshospitalet, Norway Prof. Tyge Greibrokk, Institute of Chemistry, University of Oslo Prof. Else Lundanes, Institute of Chemistry, University of Oslo Dr. Smita Ghanekar, Becton Dickinson Biosciences, California, USA Prof. Halvor Rollag, Institute of Microbiology, Rikshospitalet, Norway

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Research groups

THE FUNCTIONAL IMMUNOGENETICS GROUP GROUP LEADER: LUDVIG M. SOLLID, MD GROUP DESCRIPTION Our group is striving to understand what happens when the body's defence from disease, the immune system, directly or indirectly causes harm to the body. Coeliac disease, type 1 diabetes, rheumatoid arthritis and multiple sclerosis are examples of autoimmune disorders of a chronic inflammatory nature. People with coeliac disease get sick when they eat bread or other gluten containing food. We are concentrating on coeliac disease as a model to understand the molecular mechanisms leading to chronic inflammatory disease. In particular, we focus on how certain variants of HLA molecules predispose to disease development. There appears to be several parallels in the pathogenesis of rheumatoid arthritis and coeliac disease, and we have thus recently taken up rheumatoid arthritis as a study model. We are working in the fields of cell biology, genetics, biochemistry and protein chemistry. We are interested in the relationship between environmental factors and inherited (genetic) factors. The group is participating in EU and nationally funded projects. We are a member of the Centre for Immune Regulation (CIR) together with 4 other research groups at the University of Oslo and Rikshospitalet. This Centre, headed by Ludvig M. Solid, was in 2006 selected by the Research Council of Norway to become a Centre of Excellence. RESEARCH SUMMARY 2006 Some of the major achievements by our group in 2006 are summarised below: The inflammation of the intestinal mucosa of celiac disease is driven by gluten-reactive T cells restricted by the disease-associated HLA-DQ2 molecule. The mechanisms that regulate the activation of mucosal T cells are, however, poorly understood. Together with researchers at the Department of Pathology at Rikshospitalet, we have characterised DQ2 positive antigen presenting cells that are responsible for the activation of gluten-reactive T cells in the coeliac lesion (Ráki et al, Gastroenterology 2006). We found that DQ2+ cells in the normal duodenal mucosa consist of 2 distinct cell populations: about 80% were CD68+ DC-SIGN+ macrophages and 20% were CD11c+ dendritic cells. Importantly, the CD11c+ dendritic cells accumulate in the coeliac lesion and they express CD86 and DC-LAMP signifying their activation. When isolated from challenged biopsy specimens, we found that the CD11c+ dendritic cells efficiently activate gluten-reactive T cells. Our results suggest that a unique subset of dendritic cells is responsible for local activation of gluten-reactive T cells in the celiac lesion. T cells that are reactive with gluten peptides deamidated by transglutaminase 2 and restricted by DQ2 or DQ8 can be isolated from coeliac lesions. In an extensive study finalised in 2006, we have characterised T cell epitopes of various gliadin proteins recognised by T cells of coeliac disease patients (Tollefsen et al, J Clin Invest 2006). The results are particularly interesting, we believe, as they predict the nature of peptide epitopes that can be involved in type 1 diabetes through their presentation to diabetogenic T cells by disease associated HLA-DQ molecules. Type 1 diabetes, like celiac disease, is associated with DQ2 and DQ8 and there is a possible involvement in diabetes of trans-encoded DQ heterodimers. We found that some gliadin peptides were recognised by T cells in the context of DQ2 or DQ8 when bound in exactly the same registers, but with different requirements for deamidation; deamidation at peptide position 4 (P4) was important for DQ2-restricted T cells, whereas deamidation at P1 and/or P9 was important for DQ8-restricted T cells. Peptides combining the DQ2 and DQ8 signatures could be presented by DQ2, DQ8, and trans-encoded DQ heterodimers. It is our prediction that diabetogenic peptides will harbour these distinct signatures. Coeliac disease is a polygenic disease with involvement of HLA as well as non-HLA genes. The involved HLA genes are well defined, whereas the identity of the non-HLA genes remains largely ambiguous. Recently, a Dutch group reported in Nature Genetics paper that there is an association

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Research groups

between myosin IXB (MYO9B) gene variants and coeliac disease. We tried to replicate this finding in a cohort of Swedish and Norwegian celiac disease patients. Although our material provided reasonable power for detection, we were unable to replicate the association with the MYO9B gene polymorphism. Our effort to identify non-HLA gene predisposing to celiac disease is however continuing. Together with colleagues in Gothenburg we working hard to identify predisposing genes on chromosome 5q32-33, a region which, based on several linkage studies, are involved in coeliac disease. GROUP MEMBERS 2006 Ludvig M. Sollid (Group leader) Elin Bergseng (PhD student) Øyvind Molberg (Researcher, part time) Lars Egil Fallang (PhD student) Knut E.A. Lundin (Researcher, part time) Eirik Hornes Halvorsen (PhD student) Guro Løvik Goll (Pos Doc, part time) Ulrike Jüse (PhD student) Anders Fallang (Post Doc) Melinda Ráki (PhD student) Anna Hayman (Post Doc) Jorunn Stamnæs (PhD student) Sylvie Pollman (Post Doc, maternity leave) Marie K. Johannesen (Research Assistant) Stig Tollefsen (Post Doc) Marit Sandvik (Research Assistant) Silja S. Amundsen (PhD. student) Bjørg Simonsen (Research Assistant) Ann Christin R. Beitnes (PhD student) Ingrid Olsen (Visiting Scientist) COLLABORATIONS 2006 *National Prof. O. Bakke, Dept. of Molecular Biosciences, University of Oslo Prof. B. Bogen, Inst. Immunology, Rikshospitalet, Oslo Dr. B. Fleckenstein, Inst. Immunology, Rikshospitalet, Oslo Prof. F-E. Johansen, Institute of Pathology, Rikshospitalet, Oslo Prof I. Sandlie, Dept. of Molecular Biosciences, University of Oslo Prof. H. Scott, Institute of Pathology, Rikshospitalet HF, Oslo Ass. Prof. A.K. Uhlen, Dept. of Plant and Environ. Sciences, Norw. Univ. of Life Sciences, Ås *International Dr. H. Ascher, Dept. Paediatrics, Queen Silvia Children’s Hospital, Sahlgrenska University Hospital, Sweden Prof. S. Buus, Inst. for Medical Microbiology and Immunology, Univ. Copenhagen, Denmark Prof. L. Fugger, Weatherall Institute of Molecular Medicine, Oxford, UK Prof. C. Khosla, Dept. of Chemistry, Stanford University, USA Prof. F. Koning, Dept. Immunohematol and Blood Transfusion, Leiden Univ Medical Center, the Netherlands Assoc. Prof B. Jabri, Dept Pathology, Univ. Chicago, USA. Assoc. Prof. E. Mellins, Dept. of Pediatrics, Stanford University Prof. P. Roepstorff, Dept. Biochemistry and Molecular Biology, Univ. Southern Denmark

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Research groups

THE PROTEOMICS GROUP GROUP LEADER: BURKHARD FLECKENSTEIN, PhD LABORATORY AND GROUP Our group has continued to establish a modern mass spectrometry (MS)-based proteomics facility used to perform up-to-date research. In addition our laboratory represents the Gaustad branch of the Rikshospitalet-Radiumhospitalet Proteomics Core Facility, and it provides service to research groups at RR and UiO. The existing MALDI-TOF instrument was replaced in January 2006 for an MALDI-TOF/TOF mass spectrometer (Ultraflex II, Bruker Daltonics), and a new electrospray-Q-TOF instrument (MicroTOF-q, Bruker Daltonics) was installed in April. With our nano-liquid chromatography and spotter device we can separate small amounts of complex peptide mixtures and we get sequence information on the peptides present (LC-MS/MS). In September, two new PhD students, Ms Tutturen and Ms Dørum, have joined the proteomics group. Ms Jørgensen started in the group as a bioengineer in October. Her position is dedicated to the service provided by the Proteomics Core Facility. Our group is closely collaborating with Ludvig Sollid’s group and is part of the Sollid group constellation within the Centre of Immune Regulation (CIR). CIR was recently selected to become a Centre of Excellence by the Research Council of Norway. RESEARCH SUMMARY 2006 Our group has had its takeoff just two years ago. We are trying to understand some of the reasons why in certain diseases the immune system is directly or indirectly attacking the body itself. As model systems for such autoimmune disorders we perform research on rheumatoid arthritis (RA) and coeliac disease (CD). Patients with RA suffer from chronic inflammation of the joints which get destructed over time leading to strong motional disabilities of the patients. CD patients get sick from eating bread and other food as they do not tolerate the gluten protein present in wheat. To avoid an inflammation in the gut, they are obliged to stick to a life-long gluten free diet. Recent findings suggest that there are several parallels in the pathogenesis of RA and CD. Especially tiny changes of proteins, introduced after their synthesis by certain enzymes (post-translational modifications), have turned out to be extremely important. In RA and CD related research we use modern technologies, such as MS, to analyse proteins and peptides and to get information on these modifications. In RA, a small conversion of the amino acid arginine, called citrullination, is known to cause antibodies reacting with self proteins. Peptides carrying such citrulline residues can bind to the disease associated HLA-DR4 haplotype and may be recognized by autoreactive T cells. This modification is generated by a family of enzymes, called peptidylarginine deiminases (PAD). In collaboration with Øyvind Molberg and colleagues at IMMI we have expressed human recombinant PAD4. Ms Ekman has used a large set of synthetic peptides and MS to investigate the rules by which PAD4 selects its substrates. Understanding more about this substrate specificity might help to predict proteins which become citrullinated and therefore might be involved in the development of RA. In order to directly identify citrullinated proteins from the inflamed joints of RA patients, we have developed a chemical strategy by which citrulline residues are specifically modified (Holm et al, Anal Biochem 2006). This approach allows the specific detection of such modified citrullinated peptides by MS (Holm et al, manuscript in prep.). This technique was developed further to specifically fish out citrullinated peptides from heterogeneous mixtures (Holm, Tutturen, Fleckenstein, Patent submission in prep.). In our research on CD we closely collaborate with Ludvig Sollid who is co-supervising Ms Dørum. In CD, gluten fragments become slightly modified by an enzyme called transglutaminase type-2 (TG2). The amino acid glutamine is converted to glutamic acid (deamidation), a modification which is central for the ongoing inflammation in the intestine. Deamidated gluten peptides bind better to the disease associated HLA-DQ2 and DQ8 molecules and they are recognized by T cells in the intestine of CD patients. Ms Dørum has started her PhD project by investigating how fast TG2 can introduce deamidation into known gluten T cell epitopes. Burkhard Fleckenstein has used MS to investigate the rules by which deamidated peptides bind either to HLA-DQ2 or DQ8. We have found specific binding patterns for both alleles (Tollefsen et al, JCI 2006).

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Research groups

The Proteomics Group in October 2006. GROUP MEMBERS 2006 Burkhard Fleckenstein (Group leader) Anders Holm (Post Doc) Maria Ekman (PhD student) Astrid Tutturen (PhD student) Siri Dørum (PhD student) Marit Jørgensen (Research assistant) COLLABORATIONS 2006 Prof. L. Sollid, Inst. of Immunology Dr. Ø. Molberg, Dept. of Rheumatology and Inst of Immunology, RR HF Prof. F. Rise, Prof. K. Undheim, Dept. of Chemistry, UiO Prof I. Sandlie, Dept. of Molecular Biosciences, UiO Dr. F.E. Johansen, Inst. of Pathology, RR HF Prof. T. Tønjum, Centre for Molecular Biology and Neuroscience, Inst. of Microbiology, UiO Dr. Torgeir Holen, Prof. O.P. Ottersen, Centre for Molecular Biology and Neuroscience, Inst. of Anatomy, UiO Prof. F. Koning, Dept. Immunohematol and Blood Transfusion, Leiden Univ Medical Center, the Netherlands Prof. P. Roepstorff, Dept. of Biochemistry and Molecular Biology, Univ. Southern Denmark

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Research groups

TUMOR IMMUNOLOGY, AUTOIMMUNITY AND VACCINE DEVELOPMENT GROUP LEADER: BJARNE BOGEN, Professor MD PhD GROUP DESCRIPTION The projects in the group have one common theme: T cell recognition of peptide fragments derived from antibody molecules. Such fragments are generated by antigen processing of antibody molecules in antigen presenting cells (APC). The fragments are presented by MHC class II molecules on antigen presenting cells to T cells.

The concept of antibody molecules as antigens for T is a topic of importance for the following reasons:

1) Immune regulation and autoimmunity

2) Tumor immunology. B cell lymphomas and multiple myelomas

3) Vaccines. Troybodies

For those interested in more details, please see http://immunol.net/Projects.html

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Research groups

RESEARCH SUMMARY 2006 Research within unlinked Idiotype-dependent T-B collaboration has proceeded. This is a novel type of T-B collaboration that is distinct from conventional T-B collaboration. Using a double transgenic system established in the group, we have obtained evidence that unlinked Idiotype-dependent T-B collaboration influence production of natural IgM-like antibodies as well as IgG autoantibodies, and that it may induce lethal autoimmune disease with SLE-like features (Munthe, Zangani, Omholt, Bogen). Work on the role of Id-specific CD4+ T cells in B cell lymphomas and multiple myelomas continues with interesting results. In particular, Id-specific CD4+ T cells appear to have a curative effect on B cell lymphoma in the absence of anti-Id antibodies (Corthay, Lundin, Røsjø, Frøyland, Hofgaard, Bogen). Progress on Troybodies is good. We have devised a new screening system for defining good target specificities in the human. New T cell epitopes are being inserted into Troybodies, including telomerase. We have developed a novel type of molecules (vaccibodies) that may prove of value as a vaccine vehicle (Schjetne, Brunsvik, Tunheim, Hereng, Lindeberg, Ruffini, Bogen). GROUP MEMBERS 2006 Bjarne Bogen (Group Leader) Katrin Ulrike Lundin (PhD student) Keith Thompson (Senior Scientist) Tom Ole Løvås (Medical student) Ludvig Munthe (Post Doc) Alexandre Corthay (Post Doc) Hilde Omholt (Technician) Agnete Brunsvik Fredriksen (PhD student) Pier Ruffini (PhD student) Marianne Frøyland (PhD student) Eigil Røsjø (Medical student) Jon Magnus Haga (Medical student) Karoline Western Schjetne (Post Doc) Beate Hegge (Technician) Gro Tunheim (PhD student) Tuva Hereng (M.sci. student) Michael Zangani (PhD student) Peter Hofgaard (Technician) Lise Aagaard (Technician) Mona Lindeberg (Technician) Kristin Aas Hansen (M.sci. student) COLLABORATIONS 2006 *National Prof. I. Sandlie, Institute of Biology, University of Oslo, Oslo, Norway Prof. K. Schenck, Institute of Oral Biology, University of Oslo, Oslo, Norway Dr. G. Gaudernack, Institute of Immunology, The Norwegian Cancer Hospital, Oslo, Norway Prof. R. Blomhoff, Institute of Nutrition and Research, University of Oslo, Oslo, Norway *International Prof. L. Kwak (NIH), Bethesda, USA Prof. F. Stevenson, Tenovus laboratory, University of Southampton, England Dr. A. Grandien, Wenner-Gren Institute, Stockholm, Swede Prof. H. Yagita, Jutendo University, Japan Dr. S. Weiss, GBF, Braunschweig, Germany

23

Research groups

From top left: Bjarne Bogen Jon Magnus Haga, Michael Zangani, Katrin Lundin, Beate Hegge, Kristin Aas-Hanssen, Keith Thompson, Karoline Western Schjetne, Agnete Brunsvik Fredriksen, Marianne Frøyland, Peter Hofgaard, Hilde Omholt, Mona Lindeberg, Egil Røsjø, Alexandre Corthay, Pier Ruffini, Lise Aagaard, Gro Tunheim, Tuva Holt Hereng, Ludvig Munthe.

24

Service sections

SECTION OF MEDICAL IMMUNOLOGY (MI) HEAD: JOHANNA OLWEUS, MD (BJARTE G. SOLHEIM, MD) The section for Medical Immunology had, by the end of 2006, 11.9 positions (ASK JOHN TORGILS!) at the section (section head, 2 resident physicians and 8.9 medical technologists). The section performs immunological diagnostic analyses of samples from in- and out-patients at Rikshospitalet-Radiumhospitalet Medical Center, and from other hospitals and physicians in Norway. A novel laboraroty data system, NetLab2, was implemented in June, requiring large personnel resources for most of the year. The Institute of Immunology and Dept. of Medical Biochemistry were the first laboratories within the Laboratory Clinic to implement the system. In August, the section head, Prof. Bjarte G. Solheim, retired, and was succeeded by Johanna Olweus. To ensure a high quality of the analyses, the section participates in an international quality assessment service (UK-NEQUAS). Members of the staff have participated in research projects and teaching. SERVICE FUNCTIONS • Autoantibody testing, mainly for the diagnosis of connective tissue diseases and other

rheumatological disorders, and liver, kidney, skin,neurological diseases and celiac disease. • Complement and immunoglobulin* testing, mainly for diagnosis of lymphoproliferative disorders

and complement deficiencies, and diseases in which complement activation is involved in the pathogenesis.

• • Cellular immunology testing (quantification of subpopulations and mitogen stimulation to assess

lymphocyte proliferation), mainly for diagnosis of primary immunodeficiencies ACTIVITY 2006 (2005 IN PARENTHESIS)

78740 (70 445) • Autoantibody tests 9140 (9779)• Complement tests

• 728 (730)• Lymphocyte tests

1074 (1585)• Other types of immunological tests Altogether, the section performed 89 682 analyses. * Analyses of immunoglobulins (quantification, electrophoresis, immunofixation) have since October 2004 been performed at Dept. of Medical Biochemistry (MBK) in a collaboration between Section for Medical Immunology and MBK.

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Service sections

SECTION OF TRANSPLANTATION IMMUNOLOGY (TI) HEAD: TORSTEIN EGELAND MD (JOHN TORGILS VAAGE, MD) For most of 2006, there were 18.5 positions at the TI section (2 head physicians, 1 resident physician, 1 scientist, 14.5 biotechnologists). TI is the national centre for transplantation immunology in Norway, and provides a 24h on-call service for typing of deceased organ donors. The section is sub-divided into a serological and a genomic unit, which together cover a broad range of cellular and molecular techniques. Serological HLA typing and cross-matching is based on complement-dependent cellular cytotoxicity (CDC), antibody screening on CDC, ELISA, or multiplex flow cytometry (Luminex), while genomic HLA typing is based on sequence-specific PCR amplification (SSP), sequence-specific oligohybridization (SSO), and sequence-based typing (SBT). The TI section has accreditation from the European Federation for Immunogenetics (EFI), and has secretarial functions for the national transplantation waiting lists and the collaboration within the Nordic exchange organisation Scandia-transplant. The Norwegian Renal Registry and databases for organ transplant follow-up are administered at the section. In addition to the service functions listed below, the section focuses on certain R&D projects, such as the typing for non-HLA genes of potential importance in allotrans-plantation (e.g. Killer-cell immunoglobulin-like receptors or KIRs). Finally, The Norwegian Bone Marrow Donor Registry (NBMDR), with approx. 25,000 HLA-typed donors, is a part of TI. Please see its own annual report (www.nordonor.org). NBMDR is supported by The Norwegian Cancer Society. SERVICE FUNCTIONS • HLA-typing, HLA-antibody screening and crossmatching for allogeneic organ- and stem cell

transplantation. • HLA typing for NBMDR. • HLA typing class I for platelet transfusions for three of the national health regions. • Diagnostic HLA typing, mainly for diagnosis of Bechterew’s and Reiter’s disease, narcolepsy, celiac

disease and dermatitis herpetiformis. • (T lymphocyte counting for rejection therapy monitoring). ACTIVITY 2006 (WITH NUMBERS FROM 2005 IN PARENTHESIS)

1515 (1662)• HLA Class I typing by serology 1383 (1513)• HLA Class II typing by serology 8886 (7053)• HLA typing, genomic low resolution 4510 (3339)• HLA typing, genomic high resolution 4329 (4465)• HLA single antigen typing (disease association) 1423 (938)• Antibody screening, CDC 2003 (1887)• Antibody screening, ELISA

264 (64)• Antibody, specificity analysis 3145 (3119)• Crossmatch (CDC, B & T cells)

114 (130)• T-lymphocyte counting

171 (184)• HLA typed deceased donors (24 h on-call service)

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Service sections

ORGAN AND STEM CELL ALLOTRANSPLANTATIONS IN 2006 AT RIKSHOSPITALET-RADIUMHOSPITALET UNIVERSITY HOSPITAL

80 (87)• Kidney; living donor 125 (130)• Kidney; deceased donor

6 (10)• Kidney + pancreas 0 (1)• Pancreas, single 4 (6)• Pancreas, islets

61 (37)• Liver; deceased donor 1 (1)• Liver/ kidney

32 (28)• Heart 0 (1)• Heart/ kidney 0 (1)• Heart/ lung

20 (18)• Lung, double 4 (1)• Lung, single

31 (ND)• Stem cells, related* 26 (18)• Stem cells, unrelated*

*In addition, seven related and three unrelated donors donated stem cells to patients transplanted elsewhere.

A happy “gang” at the Section of Transplantation Immunology.

27

Teaching

Teaching Professors at the Institute teach basic and clinical immunology to medical students. The patient oriented teaching includes 1) lectures, 2) a practical course in immunological techniques including blood typing, and 3) immunologically related case stories used in "Problem based learning" , a major method of teaching at the Medical Faculty. Teaching in immunology is concentrated to the second and third year. In addition, the Institute offers postgraduate courses in immunological techniques and blood typing.

For further details about teaching of medical students, see www.med.uio.no/rh/immi. A professor at the institute is also responsible for immunology teaching at The Faculty of Mathematics and Natural Sciences, University of Oslo. Two courses are given; “Basic immunology and Immunological Techniques” and “Advanced Immunology”, both for students at the MSc and PhD levels. These courses recruit students from other Norwegian universities and university colleges. Additionally, professors at the institute have a major responsibility for teaching basic and clinical immunology to post graduate students at Oslo University College.

28

Results of our research 2006

Results of our research 2006 DOCTORAL THESES Lindenskov PHH. Inflammation in experimental meconium aspiration syndrome. Faculty of Medicine. University of Oslo. 2006. Sigurjonsson ÓE. The differentiation potential of human somatic stem cells. Faculty of Medicine. University of Oslo. 2006. Tunheim G. Recombinant antibody-based vaccines. Faculty of Medicine. University of Oslo. 2006. Wiencke K. Genetic studies in primary sclerosing cholangitis. Faculty of Medicine, University of Oslo. 2006. THESIS FOR MASTER OF SCIENCE Dahlen H. CTSL2 as a candidate gene for autoimmune diseases – genetic and functional studies to explore its role in type 1 diabetes and myasthenia gravis. University of Oslo. 2006. BOOKS AND BOOK CHAPTERS Bogen B. Lymfoproliferativ kreftsykdom og primære immundefekter. In: Klinisk biokjemi og fysiologi (Eds.: Oddva Stokke and Tr-Arne Hagve). Gyldendal Akademisk. Oslo. 2006, pp 2004-216 Lea T. Immunologi og immunologiske teknikker, Fagbokforlaget, 2006. Mollnes TE, Kirschfink M. Complement analysis in clinic and research. Advances in Experimental Medicine and Biology. 2006; 586: 361-80. Sollid LM, Molberg Ø, Lundin KEA. Celiac Disease. In: The Autoimmune Diseases, 4th edition, (Eds.: Noel I and Mackay IR,). Academic Press, Elsevier Science. London. 2006, pp 693-712. Sollid LM. Structural basis for HLA-DQ2 presentation of wheat gliadin and oat avenin to T cells. In: Coeliac Disease, Proc. of the Eleventh International Symposium on Coeliac Disease, 2004 (Eds.: McMillan S, Feighery C, Watson P, O’Farrelly C). Belfast. 2006, pp 120-125. PUBLICATIONS (Papers in peer reviewed journals, IMMI authors are underlined)

Amundsen SS1. , Monsuur AJ, Wapenaar MC, Lie BA, Ek J, Gudjonsdottir AH, Ascher H, Wijmenga C, Sollid LM. Association analysis of MYO9B gene polymorphisms with celiac disease in a Swedish/Norwegian cohort. Human Immunology. 2006; 67: 341-5.

Amundsen SS2. , Vatn M, IBSEN study group, Wijmenga C, Sollid LM, Lie BA. Association analysis of MYO9B gene polymorphisms and inflammatory bowel disease in a Norwegian cohort. Tissue Antigens. 2006; 68: 249-52.

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Results of our research 2006

Bethune MT, Strop P, Tang Y, Sollid LM3. , Khosla C. Heterologous expression, purification, refolding, and structural-functional characterization of EP-B2, a self-activating barley cysteine endoprotease. Chemistry & Biology. 2006; 13: 637-47.

4. Bjørnvold M, Amundsen SS, Stene LC, Joner G, Dahl-Jorgensen K, Njolstad PR, Ek J,

Ascher H, Gudjonsdottir AH, Lie BA, Skinningsrud B, Akselsen HE, Ronningen KS, Sollid LM, Undlien DE. FOXP3 polymorphisms in type 1 diabetes and coeliac disease. Journal of Autoimmunity. 2006; 27: 140-4.

5. Blomhoff A, Olsson M, Johansson S, Akselsen HE, Pociot F, Nerup J, Kockum I, Cambon-

Thomsen A, Thorsby E, Undlien DE, Lie BA. Linkage disequilibrium and haplotype blocks in the MHC vary in an HLA haplotype specific manner assessed mainly by DRB1*03 and DRB1*04 haplotypes. Genes and Immunity. 2006; 7: 130-40.

Bogen B6. , Ruffini P, Corthay A, Fredriksen AB, Frøyland M, Lundin K, Røsjø E, Thompson K, Massaia M. Idiotype-specific immunotherapy in multiple myeloma: suggestions for future directions of research. Haematologica. 2006; 91: 941-8.

7. Bowlus CL, Karlsen TH, Broome U, Thorsby E, Vatn M, Schrumpf E, Lie BA, Boberg KM.

Analysis of MAdCAM-1 and ICAM-1 polymorphisms in 365 Scandinavian patients with primary sclerosing cholangitis. Journal of Hepatology. 2006; 45: 704-10.

8. Braathen R, Sandvik A, Berntzen G, Hammerschmidt S, Fleckenstein B, Sandlie I,

Brandtzaeg P, Johansen FE, Lauvrak V. Identification of a Polymeric Ig Receptor Binding Phage-displayed Peptide That Exploits Epithelial Transcytosis without Dimeric IgA Competition. Journal of Biological Chemistry. 2006; 281: 7075-81.

9. Drogset JO, Grøntvedt T, Jessen V, Tegnander A, Mollnes TE, Bergh K. Comparison of in

vitro and in vivo complement activation by metallic and bioabsorbale screws used in ACL-reconstruction. Arthroscopy. 2006; 22: 489-96.

10. Franke A, Ruether A, Wedemeyer N, Karlsen TH, Nebel A, Schreiber S. No association

between the functional CARD4 insertion/deletion polymorphism and inflammatory bowel diseases in the German population. Gut. 2006; 55: 1679-80.

Fredriksen AB11. , Sandlie I, Bogen B. DNA vaccines increase immunogenicity of idiotypic tumor antigen by targeting novel fusion proteins to antigen presenting cells. Molecular Therapy 2006; 13: 776-785.

Pedersen ED12. Frøyland E, , Kvissel AK, Almaas R, Pharo A, Skålhegg BS, Mollnes TE,

Rootwelt T. Effect of acidosis on the release of IL-8 and MCP-1 during hypoxia and reoxygenation in human NT2-N neurons. Brain Research. 2006; 1113: 64-73.

13. Granum S, Sundvold-Gjerstad V, Dai KZ, Kolltveit KM, Hildebrand K, Huitfeldt HS, Lea T,

Spurkland A. Structure function analysis of SH2D2A isoforms expressed in T cells reveals a crucial role for the proline rich region encoded by SH2D2A exon 7. BMC Immunology. 2006; 13: 7-15.

Harbo HF14. , Ekstrom PO, Lorentzen AR, Sundvold-Gjerstad V, Celius EG, Sawcer S, Spurkland A. Coding region polymorphisms in T cell signal transduction genes. Prevalence and association to development of multiple sclerosis. Journal of Neuroimmunology. 2006; 177: 40-5.

Harboe M15. , Garred P, Borgen MS, Stahl GL, Roos A, Mollnes TE. Design of a complement mannose-binding lectin pathway specific activation system applicable at low serum dilutions. Clinical and Experimental Immunology. 2006; 144: 512-20.

Holm A16. , Rise F, Sessler N, Sollid LM, Undheim K, Fleckenstein B. Specific modification of peptide-bound citrulline residues. Analytical Biochemistry. 2006; 352: 68-76.

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Results of our research 2006

Holmøy T17. , Hestvik ALK, Vartdal F. Management of progressive multifocal sclerosis triggered by natalizumab. European Journal of Neurology. 2006;13: e2. (Letter to the editor)

Holmøy T18. , Hestvik ALK, Vartdal F. Towards individualised multiple sclerosis treatment. Lancet Neurology. 2006; 5: 111-2.

Holmøy T19. , Roos PM, Kvale EO. ALS: Cytokine profile of cerebrospinal fluid T cell clones. Amyotrophic Lateral Sclerosis and other Motor Neuron Disorders. 2006; 7: 183-6.

Husebye H, Halaas O, Stenmark H, Tunheim G20. , Sandanger O, Bogen B, Brech A, Latz E, Espevik T. Endocytic pathways regulate Toll-like receptor 4 signaling and link innate and adaptive immunity. EMBO Journal. 2006; 25: 683-92.

Jabri B, Sollid LM21. . Mechanisms of disease: immunopathogenesis of celiac disease. Nature Clinical Practice Gastroenterology & Hepatology. 2006; 3: 516-25.

Karlsen TH22. , Lie BA, Frey Froslie K, Thorsby E, Broome U, Schrumpf E, Boberg KM. Polymorphisms in the steroid and xenobiotic receptor gene influence survival in primary sclerosing cholangitis. Gastroenterology. 2006; 131: 781-7.

23. Kjeken R, Bogen B, Mathiesen I. DNA – fremtidens vaksine? Tidsskift for Den Norske

Lægeforening. 2006; 126: 2964-8.

Kvale EO24. , Dalgaard J, Lund-Johansen F, Rollag H, Farkas L, Midtvedt K, Jahnsen FL, Brinchmann JE, Olweus J. CD11c+ dendritic cells (DCs) and plasmacytoid DCs are activated by human cytomegalovirus and retain efficient T-cell stimulatory capability upon infection. Blood. 2006; 107: 2022-29.

25. Kveberg L, Back CJ, Dai KZ, Inngjerdingen M, Rolstad B, Ryan JC, Vaage JT, Naper C. The

novel inhibitory NKR-P1C receptor and Ly49s3 identify two complementary, functionally distinct NK cell subsets in rats. Journal of Immunology. 2006; 176: 4133-40.

26. Kveberg L, Dai KZ, Dissen E, Ryan JC, Rolstad B, Vaage JT, Naper C. Strain-dependent

expression of four structurally related rat Ly49 receptors; correlation with NK gene complex haplotype and NK alloreactivity. Immunogenetics. 2006; 58: 905-16.

Egeland T27. Lunde K, Solheim S, Aakhus S, Arnesen H, Abdelnoor M, , Endresen K, Ilebekk

A, Mangschau A, Fjeld JG, Smith HJ, Taraldsrud E, Grøgaard HK, Bjørnerheim R, Brekke M, Müller C, Hopp E, Ragnarsson A, Brinchmann JE, Forfang K. Intracoronary injection of mononuclear bone marrow cells in acute myocardial infarction. New England Journal of Medicine. 2006; 355: 1199-1209.

28. Lundmark F, Harbo HF, Celius EG, Saarela J, Datta P, Oturai A, Lindgren CM, Masterman T,

Salter H, Hillert J. Association analysis of the LAG3 and CD4 genes in multiple sclerosis in two independent populations. Journal of Neuroimmunology. 2006; 180: 193-8.

Melum E,29. Karlsen TH, Broome U, Thorsby E, Schrumpf E, Boberg KM, Lie BA. The 32-base pair deletion of the chemokine receptor 5 gene (CCR5-Delta32) is not associated with primary sclerosing cholangitis in 363 Scandinavian patients. Tissue Antigens. 2006; 68: 78-81

Molberg Ø30. , Sollid LM. A gut feeling for joint inflammation - using coeliac disease to understand rheumatoid arthritis. Trends in Immunology. 2006; 27: 188-94.

Mollnes TE31. , Kirschfink M. Strategies of therapeutic complement inhibition. Molecular Immunology. 2006; 43: 107-121.

32. Pepaj M, Holm A, Fleckenstein B, Lundanes E, Greibrokk T. Fractionation and separation of

human salivary proteins by pH gradient ion exchange and reversed phase chromatography coupled to mass spectrometry. Journal of Separation Science. 2006; 29: 519-528.

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Results of our research 2006

33. Pfeiffer-Guglielmi B, Coles JA, Francke M, Reichenbach A, Fleckenstein B, Jung G, Nicaise G, Hamprecht B. Immunocytochemical analysis of rat vagus nerve by antibodies against glycogen phosphorylase isozymes. Brain Research. 2006; 1110: 23-29.

34. Ráki M, Tollefsen S, Molberg Ø, Lundin KE, Sollid LM, Jahnsen FL . A unique dendritic cell

subset accumulates in the celiac lesion and efficiently activates gluten-reactive T cells. Gastroenterology. 2006; 131: 428-38.

Sollid LM35. , Vaage JT. Cross-dressing T cells go wild. Nature Medicine. 2006; 12: 611-2. (News and views)

36. Sprong T, Roos D, Weemaes C, Neeleman C, Gessing CLM, Mollnes TE, van Deuren M.

Deficient alternative complement pathway activation due to factor D deficiency by 2 novel mutations in the complement factor D gene in a family with meningococcal infections. Blood. 2006; 107: 4865-70.

Spurkland A, Sollid LM.37. Mapping genes and pathways in autoimmune disease. Trends in Immunology. 2006; 27: 336-42.

Sæthre M38. , Lea T, Borgen MS, Fiane AE, Michaelsen TE, Thorsby E, Haraldsen G, Mollnes TE. Human complement-activating immunoglobulin (Ig)G3 antibodies are essential for porcine endothelial cell activation. Xenotransplantation. 2006; 13: 215-23.

Thorsby E39. . Transplantation medicine in Norway through 50 years. Tidsskrift for Den Norske Laegeforening. 2006; 126: 3305-10.

Tollefsen S40. , Arentz-Hansen H, Fleckenstein B, Molberg Ø, Ráki M, Kwok WW, Jung G, Lundin KE, Sollid LM. HLA-DQ2 and -DQ8 signatures of gluten T cell epitopes in celiac disease. Journal of Clinical Investigation. 2006; 116: 2226-36.

41. Ueland T, Espevik T, Kjekshus J, Gullestad L, Omland T, Squire IB, Frøland SS, Mollnes

TE, Dickstein K, Aukrust P. Mannose binding lectin and soluble Toll-like receptor 2 in heart failure following acute myocardial infarction. Journal of Cardiac Failure. 2006; 12: 659-63.

Wiencke K42. , Boberg KM, Donaldson P, Harbo H, Ling V, Schrumpf E, Spurkland A. No major effect of the CD28/CTLA4/ICOS gene region on susceptibility to primary sclerosing cholangitis. Scandinavian Journal of Gastroenterology. 2006; 41: 586-91.

43. Wælgaard L, Pharo A, Tønnessen TI, Mollnes TE. Microdialysis for monitoring inflammation:

efficient recovery of cytokines and anaphylotoxins provided optimal catheter pore size and fluid velocity conditions. Scandinavian Journal of Immunology. 2006; 64: 345-52.

44. Xia J, Siegel M, Bergseng E, Sollid LM, Khosla C. Inhibition of HLA-DQ2-mediated antigen

presentation by analogues of a high affinity 33-residue peptide from alpha2-gliadin. Journal of the American Chemical Society. 2006; 128: 1859-67.

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Results of our research 2006

ACKNOWLEDGEMENTS The research at IMMI during 2006 continued to be extensive, as is apparent from the list of publications. A major reason for this has been the extensive collaborations between the different research groups at the institute and with other national and international groups and colleagues, which we highly acknowledge. Another major reason for the extensive research has been the support we have received from Rikshospitalet-Radiumhospitalet Medical Center and the University of Oslo, both by covering several positions and part of the running costs, and through various special allocations and grants for our research. In 2006 this also included significant financial support from Helse Sør, the national health region to which Rikshospitalet-Radiumhospitalet Medical Center belongs. Furthermore, our research depends heavily upon support from various other grants. Space does not allow us to mention all sources of support. We would like, however, to mention the extensive support from the Research Council of Norway, The Norwegian Cancer Society, The Norwegian Diabetes Society, National Institutes of Health (US), The Norwegian Council on Cardiovascular Disease, The Norwegian Foundation for Health and Rehabilitation, Medinnova, Juvenile Diabetes Foundation International, The Norwegian National Cancer Research Initiative, Gidske og Peter Jacob Sørensen's Foundation, Mr. and Mrs., Asbjørn Sognnes' Estate, The Anders Jahre Medical research Fund, The European Commission (EU), The Odd Fellow Order of Norway, The Multiple Myeloma Research Foundation (US), The Norwegian Women's Public Health Association, The Novo Nordisk Foundation, The Memorial Fund of Helga Jonsdottir and Sigurdlidi Krisjansson and The Human Frontier Science Program. To all of those who have supported our research we express our sincere gratitude.

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