Anästhesiologie
Tumor Immunology Group (TIG)
Contact/Address:
Prof. Dr. med. Jochen Greiner
Tumor-Immunology-Group (TIG)
Department of Internal Medicine III
University Hospital of Ulm
Albert-Einstein-Allee 23
89081 Ulm
Germany
E-Mail: jochen.greiner[at]uniklinik-ulm.de
Phone: | 0049+(0)731-500-45709 |
Head
Prof. Dr. med. Jochen Greiner
Project Leaders
Dr. med. Susanne Hofmann
M. Sc. Vanessa Schneider
Dr. med. Anna Babiak
Dr. med. Jacqueline Schnell
Zhang Lu
Laboratory Technicians
Marlies Götz
Cornelia Herbst
Anita Szmaragowska
Medical students
Stephanie Egenrieder
Adrian Hack
Melanie Munz
Simon Mannes
Dorothee Schönsteiner
Maximilian Steinhauser
Daria Ushmorova
Myriam Weisschuh
Anne-Katrin Wolf
Research Fields
Our research focuses on the definition of new targets for immunotherapy and on the development of vaccines for the clinical treatment of leukemia, lung cancer, renal cell carcinoma, prostate cancer and other solid tumors. Our activities include in vitro T cell assays, preclinical studies and clinical trials.
- Figure 1: Overview over the activities of the tumorimmunology group in Ulm
Identification and characterization of immunogenic targets in leukemias and solid tumors
Tumor cells can be efficiently killed by specific T cells of the immune system. Targeted immunotherapies require the identification and characterization of appropriate antigen structures. To date, a huge number of tumor-associated antigens (TAAs) have been identified and several attempts try to target these antigens to reduce tumor load or to prevent relapse in solid tumor like lung cancer, prostate cancer or renal cell carcinoma. Several enchanting immunological and even clinical responses have prompted to implement vaccination strategies to hematological malignancies like acute or chronic myeloid leukemia (AML/CML), myelodysplastic syndrome (MDS), multiple myeloma (MM) and also lymphoma. Several immunogenic antigens associated with leukemias (LAAs) have been identified in the last years in patients with hematological malignancies like BAGE, BCR-ABL, BCL-2, OFA-iLRP, FLT3-ITD, G250, hTERT, PRAME, proteinase 3, RHAMM, survivin and WT-1. Cellular and as well as humoral immune responses against these LAAs were characterized and clinical trails targeting these antigens were performed.
- Figure 2: Leukemia-associated antigens have been used in clinical trials for myeloid diseases (Aus Hofmann/Greiner, Current Cancer Drug Targets 2011).
Tumor antigens and immunological targets in solid tumors (lung cancer, renal cell carcinoma, prostate cancer)
New therapeutic strategies of lung cancer with encouraging results have been investigated including immunotherapeutic approaches, e.g. against MAGE-A3 and hTERT. Thus, tumor vaccination seems to be a promising strategy especially in situations of reduced tumor load, e.g. in maintenance therapy. In this work, we address the question, whether there are further interesting epitopes beside the antigens MAGE-A3 and hTERT inducing intensive immune reactions in a high frequency in lung cancer and are therefore eligible for immunotherapeutic approaches.
Immune reactions of CD8+ T cells were measured in ELISPOT assays for interferon gamma and granzyme B. Moreover, tetramer assays and chromium release assays were performed. Epitopes were tested derived from the lung-cancer associated antigens MAGE-A3 and hTERT, but also from the antigens RHAMM, Survivin, WT-1, PRAME, HER2 and G250 known from other tumor entities and from novel antigen structures, like Aurorakinases A and B.Most frequent responses could be found against PRAME (67%), hTERT (60%), G250 (60%) and RHAMM (40%). Lower frequency was measured for Survivin- (27%), WT-1- (27%), the two MAGE-A3- (27 and 20%) and Her2- (15%) derived peptides. Specific T cell responses could be also detected against Aurorakinases A and B. The novel peptides AuraA01 and AuraB01 showed specific T cell responses in 33% and 40% of patients respectively (Babiak/Greiner, ASCO 2011)
- Figure 3a: Immune responses of a patient with metastatic NSCLC against the antigen hTERT and with lower reactions against PRAME and MAGE (Babiak/Greiner, ASCO 2011).
Immunotargets in renal cell carcinoma and prostate cancer:
In renal cell carcinoma (RCC) T cell immune responses were also detected against different tumor antigens. The antigen RHAMM showed a high frequency of strong immune responses in patients with metastatic renal cell carcinoma (Unpublished). Therefore, the antigen RHAMM seems to be also a favourable candidate for immunotherapeutic approaches in RCC. Moreover, RHAMM expression was also detected by our group in in prostate cancer. At the protein level, RHAMM was highest in metastatic tissue samples and significantly higher in neoplastic localized disease compared with benign tissue. High RHAMM expression was associated with clinical parameters known to be linked to better clinical outcome. Summarizing, the overexpression of RHAMM at the protein level was found in both localized prostate cancer and metastatic sites. Whereas higher protein levels in primary tumors are associated with better clinical outcome, highest levels were found in lymph node metastases, an advanced stage of disease (Gust et al., Neoplasia 2009). This illustrates that RHAMM might also be a target suitable for monoclonal antibody therapy in advanced prostate cancer.
Functional role of leukemia-associated antigens (LAAs) and correlation with overall survival
- Figure 3b: LAAs are involved in different critical mechanisms relevant for cell growth. LAAs have a dual role: induction of cell proliferation but also of immune responses (Greiner et al., Clin Cancer Res 2008).
LAAs induce specific T-cell responses and are involved in crucial mechanisms for cell growth of leukemic cells. Because of this dual role, LAAs constitute exquisite target structures for targeted therapies.
Coexpression of Leukemia-associated antigens is associated with a better clinical outcome in AML patients (Figure 4a+b)
- Figure 4a: Co-expression in 116 AML patients of RHAMM, PRAME and G250 provided a favorable prognostic effect on the overall survival in AML maybe due to immune reactions (Greiner et al., Blood 2006).
- Figure 4b: Overall survival compared to mRNA expression of TAA (RHAMM, SSX2IP, Survivin). Patients with co-expression of these LAAs showed a better OS than patients without expression of these antigens (Guinn, Greiner et al., Blood 2009).
Clinical peptide vaccination trials
a) In several hematological malignancies (AML, MDS, CLL, MM)
RHAMM is an immunogenic antigen that is strongly expressed in several hematological malignancies and induces humoral and cellular immune responses. We initiated a phase I/II RHAMM-R3 peptide vaccination for patients with AML, MDS, MM and CLL overexpressing RHAMM. In this clinical study, patients with RHAMM expression but with a limited tumor load or a minimal residual disease were included. 26 patients were enrolled. The first 16 patients were vaccinated with 300 µg and further patients with 1000 µg R3 peptide.
We performed two RHAMM-R3 peptide vaccination trials using 300µg and 1000µg for patients with AML, MDS and MM overexpressing RHAMM. In the 300µg cohort we detected specific immune responses in 7/10 patients and also positive clinical effects in 5/10 patients. In the second cohort of nine patients with AML, MDS and MM vaccinated with a higher peptide dose of 1000 µg RHAMM-R3 peptide we detected specific immune responses in a lower frequency (33%) in contrast to patients in the 300µg cohort. Similar mild toxicity of both cohorts was found, only mild drug-related adverse events were observed such as erythema and induration of the skin. Nevertheless, the patients in the 300µg cohort showed a higher frequency of positive immunological clinical effects. Higher doses of peptide application might induce immune tolerance (Schmitt/Greiner Blood 2008; Greiner et al., Haematologica 2010).
Taken together, RHAMM-R3 peptide vaccination induced both immunological and clinical responses using lower and higher peptide doses. However, higher doses of peptide do not improve the frequency and intensity of immune responses in this clinical trial and might induce immune tolerance. Serological analysis and profiling is ongoing.
- Figure 5a: Synopsis of immunological and clinical data of the first 10 patients vaccinated with RHAMM 300 µg/shot (Schmitt/Greiner, Blood 2008)
- Figure 5b: Increase of RHAMM-specific T cells and decrease of Tregs of a patient with MDS vaccinated with RHAMM 1000 µg/shot after vaccination (Greiner et al., Haematologica 2010)
A further peptide vaccination trial using RHAMM-R3 peptide for patients with AML, MM, MDS is under approval and will start in 10/2011.
b) Against viral infections (CMV)
For patients after allogeneic stem cell transplantation we have initiated a peptide vaccination trial against viral CMV infection. The trial will start in 07/2011. The two most important inclusion criteria are:
• Risk constellation donor CMV negative, recipient CMV positive
(preemptive use) or
• Diagnosis of CMV infection/reactivation after allogeneic transplantation
Regulation and modulation of the immune system and immunogenicity of tumor and leukemic stem cells
We are investigating:
⇨ Selection and immunogenicity of tumor/leukemic stem cells
- Figure 6: Separation of CD34+CD38- leukemic stem cells
Frequency of Tregs in different solid tumors
⇨ Frequency of Tregs in different solid tumors
⇨ Avidity of CD8 specific T cells
⇨ Immunological mechanisms of lenalidomide and other new immunogenic drugs
⇨ Immunogenicity of modified peptides
Several immunotherapeutic Pharma-sponsored trials
Our group takes part in several immunotherapeutic clinical PST targeting tumor-associated antigens for patients with leukemias and solid tumors (see www.uniklinik-ulm.de/struktur/kliniken/innere-medizin/klinik-fuer-innere-medizin-iii.html)
Examples:
- WT-1 Vaccination in AML
- MAGE-A3 immunotherapy in lung cancer
- hTERT vaccine in lung cancer
- multi-peptide in renal cell carcinoma
Techniques
- SEREX
- RT-PCR techniques (including the quantitative light-cycler technology)
- DNA-cloning
- DNA-sequencing
- Western Blot
- Immunocytology
- Seven-color FACS analysis of surface and intracellular makers
- ELISA
- ELISPOT
- T-cell cloning, Mixed lymphocyte peptide culture (MLPC)
- 3-H proliferation assay
- 5 1-Cr release assay
- CHIP-Arrays (in Cooperation)
Grants/Funding
Deutsche Forschungsgemeinschaft (DFG), “Immunogenität mutations-spezifischer Peptidsequenzen bei der akuten myeloischen Leukämie“ Beginn ab 02/2011, 3 Jahre, Antragsteller: Prof. Dr. J. Greiner, Dr. S. Hofmann
Carreras Career Award 2010 der Deutschen José Carreras Leukämie-Stiftung e.V. „Weiterentwicklung immunologischer Behandlungsansätze für eine zielgerichtete Immuntherapie von Leukämien“. Beginn ab 09/2010. Antragsteller: Prof. Dr. J. Greiner
Deutsche José Carreras Leukämie-Stiftung e.V. „Vakzine-verstärkte Spenderlymphozyten“. Laufzeit 10/2010-09/2012. Antragsteller: Prof. Dr. J. Greiner, Prof. Dr. M. Schmitt, Dr. M. Wiesneth, Prof. Dr. H. Schrezenmeier
BMBF-Förderung des Antrages „Präventive und therapeutische Peptid-Vakzinierung gegen HCMV“ im Rahmen des Aufbaus einer Forschergruppe in der klinischen Infektiologie „Erkennung, Vermeidung und Behandlung von Infektionen des immunkompromittierten Patienten“. Projektleiter seit 01/2009: Prof. Dr. J. Greiner, Sprecher der Forschergruppe: Prof. Dr. P. Kern
Deutsche José Carreras Leukämie-Stiftung e.V. „Einfluss Leukämie-assoziierter Antigene auf die Zellproliferation myeloischer Blasten und auf das Überleben von Patienten mit akuter myeloischer Leukämie“. Beginn ab 01/2009. Antragsteller: Prof. Dr. J. Greiner, PD Dr. L. Bullinger
Else Kröner-Fresenius-Stiftung „Peptid-Vakzinierung für Patienten mit Chronischer Lymphatischer Leukämie“. Laufzeit 07/2007-06/2009. Antragsteller: Prof. Dr. M. Schmitt, PD Dr. J. Greiner, Dr. K. Giannopoulos
Deutsche José Carreras Leukämie-Stiftung e.V. „Entwicklung einer Polyvalenten Peptidvakzinierung für Patienten mit malignen hämatologischen Erkrankungen“. Laufzeit 07/2007-06/2009. Antragsteller: PD Dr. J. Greiner, Prof. Dr. M. Schmitt, Dr. M. Wiesneth, Prof. Dr. H. Schrezenmeier
Deutsche Forschungsgemeinschaft (DFG) „Rolle des Rezeptors für Hyaluronsäure-vermittelte Motilität (RHAMM/CD168) bei der Proliferation myeloischer Blasten“. Antragsteller: Dr. J. Greiner, PD Dr. M. Schmitt. Beginn der Förderung bis 07/2007
Deutsche José Carreras Leukämie-Stiftung e.V. "Charakterisierung von Antigenen bei der akuten myeloischen Leukämie". Zelluläre Assays für bereits definierte Leukämie-Antigene. Laufzeit bis einschl. 07/2005. Antragsteller: Dr. M. Schmitt, Dr. J. Greiner, Dr. M. Wiesneth, Prof. Dr. H. Döhner
Anschubfinanzierung ”Baustein-Förderung” der Universität Ulm. ”Nachweis einer spezifischen zellulären Immunantwort gegen die neu identifizierten tumor-assoziierten Antigene RHAMM, MAZ und MPP11 bei leukämischen Blasten”, Laufzeit 01/2002 bis 04/2004. Antragsteller: Dr. J. Greiner.
NOVARTIS-Stiftung, Laufzeit: 2001-2003. „Charakterisierung von immunogenen Leukämie-assoziierten Antigenen bei myeloischer Leukämien“. Antragsteller: Dr. M. Schmitt, Dr. J. Greiner.
10 Representative Publications
1. Hofmann S, Babiak A, Greiner J. Immunotherapy for Myeloproliferative Neoplasms (MPN). Curr Cancer Drug Targets 11: 72-84, 2011
IF: 5,129
2. Greiner J, Schmitt A, Giannopoulos K, Rojewski MT, Goetz M, Funk I, Ringhoffer M, Bunjes D, Hofmann S, Ritter G, Döhner H, Schmitt M. High dose RHAMM-R3 peptide vaccination for patients with acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and multiple myeloma (MM). Haematologica 95:1191-1197, 2010.
IF: 5,516
3. Guinn B*, Greiner J*, Schmitt M, and Mills K (*contributed equally). Elevated expression of the leukaemia associated antigen SSX2IP predicts good survival in acute myeloid leukaemia patients who lack detectable cytogenetic rearrangements. Blood 113: 1203-4, 2009
IF: 10,896
4. Greiner J, Guinn B, Döhner H, Bullinger L, Schmitt M. Leukemia-associated antigens (LAAs) are critical for the proliferation of acute myeloid leukemia cells. Clin Cancer Res 14: 1-6, 2008
IF: 6,250
5. Schmitt M, Schmitt A, Rojewski MT, Chen J, Giannopoulos K, Fei F, Yu Y, Götz M,Heyduk M, Ritter G, Speiser D, Gnjatic S, Guillaume P, Ringhoffer M, Schlenk RF, Liebisch P, Bunjes D, Shiku H, Döhner H, Greiner J. RHAMM-R3 peptide vaccination in patients with acute myeloid leukemia, myelodysplastic syndrome and multiple myeloma elicits immunological and clinical responses. Blood 111: 1357-1365, 2008
IF: 10,896
6. Metaxas Y, Spyridonidis A, Bertz H, Finke J, Greiner J. Donor derived mucosal epithelial cells after human hematopoietic cell transplantation are not derived from the CD34 positive fraction of the graft. Leukemia 21: 2214-16, 2007
IF: 6,924
7. Greiner J, Döhner H, Schmitt M. Cancer vaccines for patients with acute myeloid leukemia--definition of leukemia-associated antigens and current clinical protocols targeting these antigens. Haematologica 91: 1653-61, 2006
IF: 5,516
8. Greiner J*, Schmitt M*, Li Li, Giannopoulos K, Bösch K, Schmitt A, Döhner K, Schlenk RF, Pollack JR, Döhner H, Bullinger L (*contributed equally). Expression of tumor-associated antigens in acute myeloid leukemia: implications for specific immunotherapeutic approaches. Blood 108: 4109-17, 2006
IF: 10,896
9. Greiner J, Li L, Ringhoffer M, Barth T, Wiesneth M, Döhner H, Schmitt M. Identification and characterization of epitopes of the receptor for hyaluronic acid mediated motility (RHAMM/CD168) recognized by CD8 positive T cells of HLA-A2 positive patients with acute myeloid leukemia. Blood 106: 938-45, 2005
IF: 10,896
10. Greiner J, Ringhoffer M, Taniguchi M, Schmitt A, Shiku H, Döhner H, Schmitt M. mRNA expression for leukemia-associated antigens in patients with acute myeloid leukemia for the development of specific immunotherapies. Int J Cancer 108: 704-11, 2004
IF: 4,555
Collaborations
(in alphabetical order)
Thomas Barth, University of Ulm, Germany
Lars Bullinger, University of Ulm, Germany
Christian Buske, University of Ulm, Germany
Daniela Cilloni, University of Turino, Italy
Anna Dmoszynska, University of Lublin, Poland
Krzysztof Giannopoulos, University of Lublin, Poland
Barbara Guinn, University of Southampton, U.K.
Magdalena Hagn, University of Ulm, Germany
Daniel Mertens, University of Ulm, Germany
Lloyd L. Old, Ludwig Institute for Cancer Research, New York, USA
Jacek Rolinski, University of Lublin, Poland
Reinhold Schirmbeck, University of Ulm, Germany
Michael Schmitt, University of Rostock, Germany
Hiroshi Shiku, Mie University, Tsu, Japan
Markus Wiesneth, University of Ulm, Germany
Dominik Wolf, University of Innsbruck, Austria
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