Clinical Cancer Research CTRC-AACR San Antonio Breast Cancer Symposium Infection and Cancer: Biology, Therapeutics, and Prevention
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Müller, M. R.
Right arrow Articles by Brossart, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Müller, M. R.
Right arrow Articles by Brossart, P.
Clinical Cancer Research Vol. 9, 3448-3453, August 2003
© 2003 American Association for Cancer Research

Experimental Therapeutics, Preclinical Pharmacology

Expression of Her-2/neu on Acute Lymphoblastic Leukemias

Implications for the Development of Immunotherapeutic Approaches1

Martin R. Müller2, Frank Grünebach2, Katrin Kayser, Wichard Vogel, Alessio Nencioni, Wolfram Brugger, Lothar Kanz and Peter Brossart3

Department of Hematology, Oncology, and Immunology, University of Tübingen, D-72076 Tübingen, Germany


   ABSTRACT
Top
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 
Her-2/neu is a tumor-associated antigen that is expressed on several adenocarcinomas and correlates with poor prognosis. In a previous study (H. J. Bühring et al., Blood, 86: 1916–1923, 1995), it has been demonstrated that Her-2/neu expression can be detected on blast cells from patients with hematological malignancies including acute lymphoblastic leukemia (ALL). Here, we show that Her-2/neu-specific CTLs induced in vitro using peptide-pulsed dendritic cells efficiently lyse primary ALL blasts constitutively expressing both Her-2/neu and human leukocyte antigen A2 in an antigen-specific and MHC-restricted manner. Furthermore, we analyzed the feasibility of this approach in an autologous setting and induced Her-2/neu-specific CTLs using dendritic cells generated from peripheral blood mononuclear cells from an ALL patient that were pulsed with peptides or transfected with in vitro-transcribed Her-2/neu mRNA. Our data demonstrate that Her-2/neu could be used as a potential target for the application of Her-2/neu-directed treatment strategies in ALL including vaccination approaches.


   INTRODUCTION
Top
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 
Her-2/neu is a Mr 185,000 transmembrane protein with tyrosine kinase activity that is a member of the epidermal growth factor receptor family (1) . Overexpression of Her-2/neu has important biological and therapeutic implications. In preclinical studies, overexpression of Her-2/neu was found to contribute to the oncogenic transformation and metastatic potential of tumor cells (2 , 3) . In breast cancer, Her-2/neu overexpression correlates with poor prognosis and is associated with more aggressive tumors (2) . Her-2/neu is overexpressed in 20–30% of human breast cancers and can be found on several adenocarcinomas such as ovarian, colorectal, or RCCs4 (2, 3, 4) . Her-2/neu protein currently represents a viable therapeutic target for the development of novel treatment strategies using mAbs or tyrosine kinase inhibitors that have been shown to be effective in the treatment of malignant diseases (3 , 5, 6, 7) . Furthermore, in recent vaccination studies, DCs presenting T-cell epitopes derived from the Her-2/neu oncogene were demonstrated to induce antigen-specific CTLs in cancer patients that are able to recognize tumor cells expressing Her-2/neu (8) . In a previous report (9) , it was found that Her-2/neu is expressed in some hematological malignancies including ALL. To further extend the possible use of Her-2/neu-derived T-cell epitopes in immunotherapeutic approaches, we analyzed the presentation of antigenic peptides by these malignant cells using in vitro-induced Her-2/neu-specific CTLs. CTLs generated from healthy donors by primary in vitro immunization recognized malignant cells from patients with ALL in an antigen-specific manner. Furthermore, we show that Her-2/neu-specific CTLs able to lyse autologous malignant cells can be induced in patients with ALL using peptide-pulsed or RNA-transfected DCs, indicating that functional DCs and CTLs can be generated in these patients. Our results demonstrate that Her-2/neu is a tumor-associated antigen expressed in ALL cells and contribute to the development of novel therapeutic strategies in ALL.


   MATERIALS AND METHODS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS AND DISCUSSION
 REFERENCES
 
Tumor Cell Lines.
Tumor cell lines used in the experiments were grown in RP10 medium (4) . The following tumor cells were used in the experiments: A498 (RCC; Her-2/neu+; HLA-A2+); Croft [EBV-immortalized B-cell line kindly donated by O. J. Finn (Pittsburgh, PA); Her-2/neu-; HLA-A2+]; SK-OV-3 (ovarian carcinoma cell line; Her-2/neu+; HLA-A3+; kindly provided by O. J. Finn); and K562 (chronic myelogeneous leukemia cell line).

Blasts from ALL Patients.
Blasts from seven ALL patients were used for the experiments: (a) patient 1, HLA-A2+, Her-2/neu+; (b) patient 2, HLA-A2+, Her-2/neu+; (c) patient 3, HLA-A2-, Her-2/neu+; (d) patient 4, HLA-A2+, Her-2/neu+; (e) patient 5, HLA-A2+, Her-2/neu+; (f) patient 6, HLA-A2-, Her-2/neu-; and (g) patient 7, HLA-A2+, Her-2/neu-. Blasts from patients with ALL were grown in RP10 medium for 24 h before they were used as target cells in a standard 51Cr release assay.

Generation of DCs.
Generation of DCs from adherent PBMNCs was performed as described previously (4 , 10) .

Cell Isolation.
B cells and CD34+ peripheral blood progenitor cells were isolated using the B Cell Isolation Kit (human) and the CD34+ Progenitor Cell Isolation Kit [human (Miltenyi Biotec, Bergisch Gladbach, Germany)] according to the manual provided by the manufacturer. Isolated cell populations were characterized by flow cytometry.

Immunostaining.
Cell immunostaining and detection of Her-2/neu expression were performed as described elsewhere (4 , 8 , 10 , 11) .

Induction of an Antigen-specific CTL Response using HLA-A2-restricted Synthetic Peptides and CTL Assay.
CTL induction with the Her-2/neu-derived peptides [E75, KIFGSLAFL (12) ; GP2, IISAVVGIL (13) ] and the 51Cr release assay for CTL activity were performed as described previously (4 , 10 , 14, 15, 16) . Briefly, DCs were pulsed with 50 µg/ml synthetic peptide for 2 h, washed, and incubated with 3 x 106 autologous PBMNCS in RP10 medium. After 7 days and 14 days of culture, cells were restimulated with autologous peptide-pulsed PBMNCS, and 1 ng/ml human recombinant IL-2 (R&D Systems, Wiesbaden, Germany) was added every second day after the first restimulation. The cytolytic activity of induced CTLs was analyzed on day 19 (day 5 after the last restimulation) in a standard 51Cr release assay. Antigen specificity of tumor cell lysis was further determined in a cold target inhibition assay (4) by analyzing the capacity of peptide-pulsed unlabeled Croft cells to block the lysis of tumor cells at a ratio of 20:1 (inhibitor:target ratio) and by blocking studies. For antibody blocking, target cells were incubated for 30 min with 20 µg/ml mAb BB7.2 recognizing the HLA-A2 molecule or isotype control antibody. For preincubation with IFN-{gamma}, blasts were cultured for 24 h in RP10 medium with 100 units/ml IFN-{gamma} (Genzyme, Neu-Isenburg, Germany) before the assay.

Generation of Her-2/neu mRNA by in Vitro Transcription.
For generation of Her-2/neu mRNA, plasmid pSPJC1 with an insert coding for Her-2/neu (kindly provided by Prof. A. Ulrich, Martinsried, Germany) which allows in vitro transcription under the control of a SP6 promoter, was used. The plasmid was linearized using NdeI (MBI Fermentas, St. Leon-Rot, Germany) and in vitro transcribed with SP6 Cap Scribe (Roche, Mannheim, Germany) according to the protocol provided by the manufacturer. Purification of in vitro transcripts was performed with RNeasy Mini anion-exchange spin columns (Qiagen, Hilden, Germany) according to the RNA cleanup protocol provided by the manufacturer. In vitro transcripts were routinely checked by formaldehyde/agarose gel electrophoresis for size and integrity and stored at -80°C in small aliquots.

Electroporation of DCs and CTL Induction.
Before electroporation on day 6, immature DCs were washed twice with serum-free X-VIVO 20 medium (BioWhittaker, Walkersville, MD) and resuspended to a final concentration of 2 x 107 cells/ml. Subsequently, 200 µl of the cell suspension were mixed with 5 µg of Her-2/neu mRNA and electroporated in a 4-mm cuvette using an Easyject Plus unit (Peqlab, Erlangen, Germany; Ref. 17 ). The physical parameters were voltage of 300 V, capacitance of 150 µF, resistance of 1540 {Omega}, and pulse time of 231 ms. After electroporation, the cells were immediately transferred into RP10 medium and returned to the incubator. After transfection, DCs were incubated for 24 h in RP10 medium containing 10 ng/ml TNF-{alpha} for maturation of DCs. For CTL induction, 5 x 105 DCs (transfected with Her-2/neu RNA) were coincubated with 2.5 x 106 autologous PBMNCS in RP10 medium. Human recombinant IL-2 (1 ng/ml; Genzyme) was added on days 7, 9, and 11. The cytolytic activity of induced CTLs was analyzed on day 11 after CTL induction in a standard 51Cr release assay.


   RESULTS AND DISCUSSION
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
REFERENCES
 
Her-2/neu Expression on Primary ALL Blasts.
We analyzed the expression of Her-2/neu on primary blasts from patients with ALL using a Her-2/neu-specific monoclonal antibody. In 5 of 33 tested patients with ALL, expression of Her-2/neu was detected. Interestingly, all five patients had a poor outcome with early relapse or resistance to standard treatment. As demonstrated in Fig. 1ACitation , ALL blasts were characterized by high levels of CD10, CD19, CD34, and HLA-DR expresssion. In Fig. 1BCitation , the level of Her-2/neu expression on ALL blasts was assessed by analyzing its presence on CD10+ and CD19+ lymphocytes. Table 1Citation demonstrates the range of Her-2/neu expression on blasts of all five Her-2/neu+ patients displayed as the mean fluorescence intensity as assessed by flow cytometry.



View larger version (51K):
[in this window]
[in a new window]
  		Fig. 1. Flow cytometric analysis of Her-2/neu expression on ALL blasts. Blast cells were characterized by immunostaining for typical ALL markers such as CD34, HLA-DR, CD10, and CD19 as demonstrated for a representative patient (patient 1, gated on the lymphocyte population; A). The expression of Her-2/neu on CD10+ and CD19+ ALL blasts is shown in B for two Her-2/neu+ patients (patients 3 and 5) and two Her-2/neu- patients (patients 6 and 7).

 

View this table:
[in this window]
[in a new window]
  		Table 1 Mean fluorescence intensity of Her-2/neu + ALL blasts

 
Her-2/neu-specific CTLs Generated from PBMNCs of Healthy Donors Recognize Primary ALL Blasts.
To analyze the presentation of Her-2/neu antigenic peptides by primary ALL cells, we induced Her-2/neu-specific CTLs in vitro using peptide-pulsed DCs generated from PBMNCS of HLA-A2+ healthy donors as antigen-presenting cells (4 , 10) . The two recently described HLA-A2-binding peptides E75 (12) and GP2 (13) were used for CTL induction in vitro (4) . Croft cells (HLA-A2+/Her-2/neu-) were lysed when they were pulsed with the cognate peptide used for CTL induction but were spared when they were pulsed with an irrelevant peptide or left untreated. As shown in Fig. 2ACitation , CTL lines obtained after two weekly restimulations demonstrated peptide-specific killing. Both CTL lines were able to efficiently lyse A498 cells (HLA-A2+/Her-2/neu+), whereas there was no lysis of the ovarian cancer SK-OV-3 cells (Her-2/neu+/HLA-A3+). There was no killing of K562 cells (Fig. 2B)Citation , demonstrating that the cytotoxic activity was not mediated by NK cells. The cytolytic avtivity against RCC line A498 could be blocked entirely by using a monoclonal antibody against HLA-A2. These results demonstrate that the presentation of Her-2/neu peptides in the context of HLA-A2 molecules on the target cells is necessary for the efficient lysis of tumor cells and confirm the antigen specificity and MHC restriction of the induced CTLs. As shown in Fig. 2CCitation , the Her-2/neu-specific CTLs did not recognize autologous nonmalignant B cells, DCs, bone marrow, and mobilized CD34+ hematopoietic progenitor cells from a healthy HLA-A2+ donor, whereas there was a significant lysis of leukemic blasts from a HLA-A2+ and Her-2/neu+ ALL patient. These results demonstrate that the induced cytolytic activity is directed specifically against the Her-2/neu-expressing ALL blasts.



View larger version (28K):
[in this window]
[in a new window]
  		Fig. 2. Cytotoxic specificity of CTLs induced with Her-2/neu peptide-pulsed DCs from healthy donors. DCs were generated from adherent PBMNCs grown for 7 days in RP10 medium supplemented with GM-CSF, IL-4, and TNF-{alpha} and pulsed with the synthetic Her-2/neu-derived peptides E75 (left column) or GP2 (right column) and used as antigen-presenting cells to induce a CTL response in vitro. The cytotoxic activity of induced Her-2/neu-specific CTLs (CTL E75 and CTL GP2) was determined in a standard 51Cr release assay after two weekly restimulations. Targets used were the EBV-immortalized B-cell line Croft (Her-2/neu-, HLA-A2+) pulsed with the Her-2/neu-derived peptides E75 and GP2, the RCC cell line A498 (Her-2/neu+, HLA-A2+), and the ovarian carcinoma cell line SK-OV-3 [Her-2/neu+, HLA-A2- (A)]. To rule out NK-mediated cytotoxicity, K562 cells were also included as target cells. A498 cells were incubated with a HLA-A2-specific mAb (BB7.2) before the assay and used as target cells to show the HLA-A2 restriction of the CTLs. A498 cells incubated with mouse IgG were included as an isotype control (B). The cytotoxic activity of the two CTL lines was further tested against ALL blasts from patient 5 (Her-2/neu+, HLA-A2+) as well as against nonmalignant B cells, DCs, bone marrow and CD34+ peripheral hematopoietic progenitor cells (all HLA-A2+) (C).

 
In the next set of experiments, we analyzed the ability of the in vitro-induced CTLs to lyse primary leukemic cells from patients with ALL. Flow cytometric analysis revealed Her-2/neu expression on blasts obtained from five patients with ALL (Fig. 1Citation and Table 1Citation ). As shown in Fig. 3ACitation , Her-2/neu peptide-specific CTLs recognized primary Her-2/neu-expressing ALL blasts obtained from three HLA-A2+ patients (patients 1, 2, and 4), suggesting that Her-2/neu peptides are presented by these leukemic cells. In contrast, there was no lysis of cells missing the corresponding HLA haplotype (HLA-A2-, patient 3). The antigen specificity mediated by the in vitro-induced CTL lines was further confirmed in a cold target inhibition assay (Fig. 3B)Citation . The lysis of the ALL cells (patient 1) could be blocked by the addition of unlabeled "cold" Croft cells pulsed with the cognate peptide, whereas cells pulsed with an irrelevant peptide showed no effect. In our experiments, we observed a diminished lysis of primary ALL blasts from patient 2 that was not due to a decreased expression of Her-2/neu protein on the ALL blasts (Fig. 4A)Citation . We therefore analyzed the expression of HLA class I molecules on the surface of these cells, and it was reduced as compared with the blasts from the other patients and could be up-regulated by incubation with IFN-{gamma}. In line with these results, the lysis of the ALL blasts of patient 2 by the CTLs could be increased upon treatment of the target cells with IFN-{gamma} (Fig. 4B)Citation .



View larger version (25K):
[in this window]
[in a new window]
  		Fig. 3. Lysis of ALL blasts by Her-2/neu-specific CTLs induced with peptide-pulsed DCs from healthy donors. To analyze the ability of Her-2/neu-specific CTLs to lyse primary ALL blasts, ALL cells from four different ALL patients were used as targets in a standard 51Cr release assay [patient 1, Her-2/neu+, HLA-A2+; patient 2, Her-2/neu+, HLA-A2+; patient 3, Her-2/neu+, HLA-A2-; patient 4, Her-2/neu+, HLA-A2+ (A)]. Cytolytic activity of CTLs could be inhibited by cold (unlabeled) Croft cells pulsed with the cognate peptide but not by cold (unlabeled) Croft cells pulsed with an irrelevant peptide (B).

 


View larger version (19K):
[in this window]
[in a new window]
  		Fig. 4. Influence of IFN-{gamma} on the lysis of ALL blasts. The level of Her-2/neu protein expression on ALL blasts from patient 2 was assessed by immunostaining using a phycoerythrin-conjugated antibody (A). To test the possible influence of IFN-{gamma} on the cytolytic activity of the CTLs, blasts from patient 2 were preincubated for 24 h with IFN-{gamma} and used as targets in a standard 51Cr release assay. CTL-E75 cells were used as effector cells (B).

 
Her-2/neu-specific CTLs Can Be Induced using PBMNCs from ALL Patients.
Next we analyzed the induction of Her-2/neu-specific CTLs in an autologous setting using PBMNCS from a patient (patient 5) with a newly diagnosed Her-2/neu+ ALL. DCs were generated from peripheral blood monocytes of this patient and pulsed with the GP2 and E75 peptides. In addition, these DCs were transfected with an in vitro-transcribed Her-2/neu mRNA and used as antigen-presenting cells for CTL induction. The cytotoxic activity of the T cells generated with RNA-transfected DCs was determined on day 11, whereas the peptide-induced CTLs were analyzed on day 19 after additional weekly restimulation with peptide-pulsed autologous PBMNCS.

As demonstrated in Fig. 5ACitation , CTLs generated with Her-2/neu RNA-transfected DCs lysed HLA-A2-matched allogeneic tumor cells and autologous malignant ALL cells. There was no significant cytolytic activity directed against K562 cells, demonstrating that the induced response was not mediated by NK cells. Interestingly, the CTLs induced with Her-2/neu RNA-transfected DCs also recognized targets pulsed with the E75 or GP2 peptides, demonstrating that these two peptides are presented upon transfection of DCs with Her-2/neu RNA and contribute to the observed cytolytic activity.



View larger version (19K):
[in this window]
[in a new window]
  		Fig. 5. Induction of Her-2/neu-specific CTLs with RNA-transfected or peptide-pulsed DCs from ALL patients. DCs were generated using adherent PBMNCs from a patient with a newly diagnosed ALL. Autologous DCs were cultured in RP10 medium supplemented with GM-CSF and IL-4 and electroporated with Her-2/neu mRNA using an Easyject Plus unit on day 6 after induction. After an additional 24 h in RP10 medium supplemented with GM-CSF, IL-4, and TNF-{alpha}, DCs were used for CTL induction. Cytolytic activity was determined by a standard 51Cr release assay on day 11 after CTL induction (A). DCs used for peptide pulsing (B) were prepared as described in the previous experiments. Cytolytic activity was analyzed by a standard 51Cr release assay after two weekly restimulations with peptide-pulsed autologous PBMNCs. Target cells used in the assays were autologous ALL blasts (Her-2/neu+, HLA-A2+), the EBV-immortalized B-cell line Croft (Her-2/neu-, HLA-A2+) pulsed with the Her-2/neu-derived peptides E75 or GP2 or with the irrelevant HIV peptide, the RCC cell line A498 (Her-2/neu+, HLA-A2+), and the ovarian cancer carcinoma line SK-OV-3 (Her-2/neu+, HLA-A2-). To assess for potential NK cell-mediated cytotoxicity, the K562 cell line was included.

 
Lysis of autologous malignant cells was also detected when CTLs generated with peptide-pulsed DCs were applied in the assay (Fig. 5B)Citation . However, these CTLs required additional restimulation, indicating that transfection of DCs with RNA derived from Her-2/neu is a more potent approach to elicit tumor-specific CTLs.

In our study, we demonstrate that the tumor-associated antigen Her-2/neu is expressed in ALL and that primary blasts from patients with ALL can be lysed by CTLs specific for Her-2/neu-derived peptides. In a previous report, it was shown that Her-2/neu is expressed in a subset of hematological malignancies, especially in CD10+/CD19+ common-ALL (in 9 of 19 patients; Ref. 9 ) indicating that treatment strategies targeting Her-2/neu could be evaluated in this subpopulation of patients. Recent developments in the treatment of malignant diseases using mAbs, tyrosine kinase inhibitors, or cellular immunotherapies have revealed Her-2/neu as an important therapeutic target (3 , 5, 6, 7, 8) .

In contrast to chronic myelogenous leukemia, where remissions of the disease can be induced in 60–80% of patients after donor lymphocyte infusions, such clinical and immunological effects are rare in ALL patients due to the induction of a graft-versus-leukemia effect (18 , 19) . This might be due to a reduced induction of ALL-specific T cells after donor lymphocyte infusions or might result from inefficient recognition of ALL blasts by the CTLs. However, our data indicate that ALL-specific T cells can be generated in the peripheral blood of patients with ALL in vitro and that these CTLs are able to efficiently lyse primary autologous leukemia cells. Thus, enhanced antigen presentation by a powerful antigen-presenting cell (such as DCs) might induce a strong T-cell response capable of mediating the lysis of ALL blasts, which could result in clinical remission of the leukemic disease.

In a Phase I study using DCs pulsed with HLA-A2-binding peptides derived from Her-2/neu or MUC1 tumor antigens (8) , we were recently able to induce peptide-specific CTLs in patients with metastatic breast and ovarian cancers in vivo without any side effects or autoimmune reactions, demonstrating that these peptides can be safely and efficiently used in clinical studies.

In conclusion, our results suggest that therapies targeting Her-2/neu, including vaccination studies with peptide-pulsed DCs, could be applied in ALL.


   ACKNOWLEDGMENTS
 
We thank Bruni Schuster and Sylphia Stephan for excellent technical assistance.


   FOOTNOTES
 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 Supported in part by grants from the Deutsche Forschungsgemeinschaft (SFB 510), Deutsche Krebshilfe, and the Fortuene Program of the University of Tübingen. Back

2 Both authors contributed equally to this work. Back

3 To whom requests for reprints should be addressed, at Department of Hematology, Oncology, and Immunology, University of Tübingen, Otfried-Müller-Strasse 10, D-72076 Tübingen, Germany. Phone: 49-7071-2982726; Fax: 49-7071-293671; E-mail: peter.brossart{at}med.uni-tuebingen.de Back

4 The abbreviations used are: RCC, renal cell carcinoma; mAb, monoclonal antibody; DC, dendritic cell; ALL, acute lymphoblastic leukemia; HLA, human leukocyte antigen; PBMNC, peripheral blood mononuclear cell; NK, natural killer; IL, interleukin; TNF, tumor necrosis factor; GM-CSF, granulocyte macrophage colony-stimulating factor. Back

Received 11/18/02; revised 4/ 9/03; accepted 4/10/03.


   REFERENCES
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 

  1. Yamamoto T., Ikawa S., Akiyama T., Semba K., Nomura N., Miyajima N., Saito T., Toyoshima K. Similarity of protein encoded by the human c-erb-B-2 gene to epidermal growth factor receptor. Nature (Lond.), 319: 230-234, 1986.[CrossRef][Medline]
  2. Slamon D. J., Godolphin W., Jones L. A., Holt J. A., Wong S. G., Keith D. E., Levin W. J., Stuart S. G., Udove J., Ullrich A. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science (Wash. DC), 244: 707-712, 1989.[Abstract/Free Full Text]
  3. Wang S. C., Hung M. C. HER2 overexpression and cancer targeting. Semin. Oncol., 28 (Suppl. 16): 115-124, 2001.[CrossRef][Medline]
  4. Brossart P., Stuhler G., Flad T., Stevanovic S., Rammensee H. G., Kanz L., Brugger W. Her-2/neu-derived peptides are tumor-associated antigens expressed by human renal cell and colon carcinoma lines and are recognized by in vitro induced specific cytotoxic T lymphocytes. Cancer Res., 58: 732-736, 1998.[Abstract/Free Full Text]
  5. Yip Y. L., Ward R. L. Anti-ErbB-2 monoclonal antibodies and ErbB-2-directed vaccines. Cancer Immunol. Immunother., 50: 569-587, 2002.[CrossRef][Medline]
  6. Moasser M. M., Basso A., Averbuch S. D., Rosen N. The tyrosine kinase inhibitor ZD1839 ("Iressa") inhibits HER2-driven signaling and suppresses the growth of HER2-overexpressing tumor cells. Cancer Res., 61: 7184-7188, 2001.[Abstract/Free Full Text]
  7. Sawyers C. L. Rational therapeutic intervention in cancer: kinases as drug targets. Curr. Opin. Genet. Dev., 12: 111-115, 2002.[CrossRef][Medline]
  8. Brossart P., Wirths S., Stuhler G., Reichardt V. L., Kanz L., Brugger W. Induction of cytotoxic T-lymphocyte responses in vivo after vaccinations with peptide-pulsed dendritic cells. Blood, 96: 3102-3108, 2000.[Abstract/Free Full Text]
  9. Bühring H. J., Sures I., Jallal B., Weiss F. U., Busch F. W., Ludwig W. D., Handgretinger R., Waller H. D., Ullrich A. The receptor tyrosine kinase p185HER2 is expressed on a subset of B-lymphoid blasts from patients with acute lymphoblastic leukemia and chronic myelogeneous leukemia. Blood, 86: 1916-1923, 1995.[Abstract/Free Full Text]
  10. Brossart P., Heinrich K. S., Stuhler G., Behnke L., Reichardt V. L., Stevanovic S., Muhm A., Rammensee H. G., Kanz L., Brugger W. Identification of HLA-A2-restricted T-cell epitopes derived from the MUC1 tumor antigen for broadly applicable vaccine therapies. Blood, 93: 4309-4317, 1999.[Abstract/Free Full Text]
  11. Bühring H. J., Seiffert M., Marxer A., Weiss B., Faul C., Kanz L., Brugger W. AC133 antigen expression is not restricted to acute myeloid leukemia blasts but is also found on acute lymphoid leukemia blasts and on a subset of CD34+ B-cell precursors. Blood, 94: 832-833, 1999.[Free Full Text]
  12. Fisk B., Blevins T. L., Wharton J. T., Ioannides C. G. Identification of an immunodominant peptide of HER-2/neu protooncogene recognized by ovarian tumor-specific cytotoxic T lymphocyte lines. J. Exp. Med., 181: 2109-2717, 1995.[Abstract/Free Full Text]
  13. Peoples G. E., Goedegebuure P. S., Smith R., Linehan D. C., Yoshino I., Eberlein T. J. Breast and ovarian cancer specific cytotoxic T lymphocytes recognize the same HER-2/-neu derived peptide. Proc. Natl. Acad. Sci. USA, 92: 432-436, 1995.[Abstract/Free Full Text]
  14. Brossart P., Zobywalski A., Grunebach F., Behnke L., Stuhler G., Reichardt V. L., Kanz L., Brugger W. Tumor necrosis factor {alpha} and CD40 ligand antagonize the inhibitory effects of interleukin 10 on T-cell stimulatory capacity of dendritic cells. Cancer Res., 60: 4485-4492, 2000.[Abstract/Free Full Text]
  15. Brossart P., Bevan M. J. Selective activation of Fas/Fas ligand-mediated cytotoxicity by a self peptide. J. Exp. Med., 183: 2449-2458, 1996.[Abstract/Free Full Text]
  16. Brossart P., Schneider A., Dill P., Schammann T., Grünebach F., Wirths S., Kanz L., Brugger W. The epithelial mucin MUC1 is expressed in hematological malignancies and is recognized by MUC1 specific cytotoxic T-lymphocytes. Cancer Res., 61: 6846-6850, 2001.[Abstract/Free Full Text]
  17. Grünebach F., Müller M. R., Nencioni A., Brossart P. Delivery of tumor derived RNA for the induction of cytotoxic T-lymphocytes. Gene Ther., 10: 367-374, 2003.[CrossRef][Medline]
  18. Van Rhee F., Kolb H. J. Donor leukocyte transfusions for leukemic relapse. Curr. Opin. Hematol., 2: 423-430, 1995.[Medline]
  19. Luznik L., Fuchs E. J. Donor lymphocyte infusions to treat hematologic malignancies in relapse after allogeneic blood or marrow transplantation. Cancer Control, 9: 123-137, 2002.[Medline]



This article has been cited by other articles:


Home page
 Clin. Cancer Res.Home page
C. N. Boss, F. Grunebach, K. Brauer, M. Hantschel, V. Mirakaj, T. Weinschenk, S. Stevanovic, H.-G. Rammensee, and P. Brossart
Identification and Characterization of T-Cell Epitopes Deduced from RGS5, a Novel Broadly Expressed Tumor Antigen
Clin. Cancer Res., June 1, 2007; 13(11): 3347 - 3355.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
S. Maia, W. N. Haining, S. Ansen, Z. Xia, S. A. Armstrong, N. P. Seth, P. Ghia, M. L. den Boer, R. Pieters, S. E. Sallan, et al.
Gene Expression Profiling Identifies BAX-{delta} as a Novel Tumor Antigen in Acute Lymphoblastic Leukemia
Cancer Res., November 1, 2005; 65(21): 10050 - 10058.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
M. R. Muller, G. Tsakou, F. Grunebach, S. M. Schmidt, and P. Brossart
Induction of chronic lymphocytic leukemia (CLL)-specific CD4- and CD8-mediated T-cell responses using RNA-transfected dendritic cells
Blood, March 1, 2004; 103(5): 1763 - 1769.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Müller, M. R.
Right arrow Articles by Brossart, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Müller, M. R.
Right arrow Articles by Brossart, P.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online