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:
Fax:

0049+(0)731-500-45709
0049+(0)731-500-45750

Project Leaders

Prof. Dr. med. Jochen Greiner

Dr. med. Susanne Hofmann

Dr. med. Anna Babiak

Laboratory Technicians

Marlies Götz

Cornelia Herbst

Anita Szmaragowska

Medical students

Stephanie Egenrieder

Adrain Hack

Marta Heyduk

Lena Kienle

Simon Mannes

Dorothee Sperlich

Maximilian Steinhauser

Daria Ushmorova

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 and urological 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

1. 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: Several Leukemia-associated antigens have been identified in different hematological diseases

2. Functional role of leukemia-associated antigens (LAAs) and correlation with overall survival

: Figure 3: 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)

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).

3. Clinical peptide vaccination trials

- 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 were included with RHAMM expression but with a limited tumor load or a minimal residual disease. 26 patients were enrolled. The first 16 patients were vaccinated with 300 mcg and further patients with 1000 mcg 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 in 7/10 patients specific immune responses 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.

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.

: Figure 5: Synopsis of immunological and clinical data of the first 10 patients vaccinated with RHAMM 300 µg/shot (Schmitt/Greiner, Blood 2008)

A further peptide vaccination trial using RHAMM-R3 peptide for patients with AML, MM, MDS is under approval and will start at 06/2010

- Against viral infections (CMV)

For patients after allogeneic stem cell transplantation we have initiated a peptide vaccination trial against a viral CMV infection. The trial will start at 03/2010. 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

Primary Aims:

• Safety and feasibility of vaccination with CMV-specific peptide after allogeneic bone marrow or peripheral stem cell transplantation

Secondary Aims:

• Induction of a CMV-specific T-cell response

• Generation of a enduring, protective immune response against CMV and thereby reduction of the CMV-reactivation in patients after allogeneic bone marrow or peripheral stem cell transplantation

4. Tumor antigens and immunological targets in solid tumors (lung cancer, renal cell carcinoma, prostate cancer)

Aims are:

⇨ Investigation of specific T cell responses against known or new LAAs in NSCLC and SCLC

⇨ Examination of specific T cell responses against LAAs by Tetramer arrays in different solid tumors (Cooperation with B. Guinn, Southampton)

⇨ Correlation of survival and RHAMM expression in RCC and prostate cancer (Gust et al., Neoplasia 2009)

5. Regulation and modulation of the immune system

We are investigating:

⇨ Frequency of Tregs

⇨ Avidity of CD8 specific T cells

⇨ Immunological mechanisms of lenalidomide

⇨ Immunogenicity of modified peptides

6. 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)

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

Grants/Funding

NOVARTIS-Stiftung, Laufzeit: 2001-2003. „Charakterisierung von immunogenen Leukämie-assoziierten Antigenen bei myeloischer Leukämien“. Antragsteller: Dr. M. Schmitt, Dr. J. Greiner.

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.

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,

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. „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

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

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

Representative Publications

1. 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, in press, 2010.

IF: 5,516

2. 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

3. 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

4. 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

5. 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

6. 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

7. 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

8. 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

9. 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 immuno¬therapies. Int J Cancer 108: 704-11, 2004

IF: 4,555

10. Greiner J, Ringhoffer M, Taniguchi M, Hauser T, Schmitt A, Döhner H, Schmitt M.

Characterization of several leukemia-associated antigens inducing humoral immune responses in acute and chronic myeloid leukemia (AML/CML). Int J Cancer 106: 224-31, 2003

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