Publications by authors named "Peter I Schrier"

8 Publications

  • Page 1 of 1

Detection and functional analysis of CD8+ T cells specific for PRAME: a target for T-cell therapy.

Clin Cancer Res 2006 May;12(10):3130-6

Department of Clinical Oncology, Leiden University Medical Center, Leiden, Netherlands.

Purpose: Preferentially expressed antigen on melanomas (PRAME) is an interesting antigen for T-cell therapy because it is frequently expressed in melanomas (95%) and other tumor types. Moreover, due to its role in oncogenic transformation, PRAME-negative tumor cells are not expected to easily arise and escape from T-cell immunity. The purpose of this study is to investigate the usefulness of PRAME as target for anticancer T-cell therapies.

Experimental Design: HLA-A*0201-subtyped healthy individuals and advanced melanoma patients were screened for CD8+ T cells directed against previously identified HLA-A*0201-binding PRAME peptides by IFN-gamma enzyme-linked immunosorbent spot assays and tetramer staining. PRAME-specific T-cell clones were isolated and tested for recognition of melanoma and acute lymphoid leukemia (ALL) cell lines. PRAME mRNA expression was determined by quantitative real-time reverse transcription-PCR.

Results: In 30% to 40% of healthy individuals and patients, PRA(100-108)-specific CD8+ T cells were detected both after in vitro stimulation and directly ex vivo after isolation by magnetic microbeads. Although CD45RA- memory PRA(100-108)-specific T cells were found in some individuals, the majority of PRA(100-108)-tetramer+ T cells expressed CD45RA, suggesting a naive phenotype. PRA(100-108)-tetramer+ T-cell clones were shown to recognize and lyse HLA-A*0201+ and PRAME+ melanoma but not ALL cell lines. Quantitative real-time reverse transcription-PCR showed significantly lower PRAME mRNA levels in ALL than in melanoma cell lines, suggesting that PRAME expression in ALL is below the recognition threshold of our PRA(100-108)-tetramer+ T cells.

Conclusion: These data support the usefulness of PRAME and in particular the PRA(100-108) epitope as target for T-cell therapy of PRAME-overexpressing cancers.
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http://dx.doi.org/10.1158/1078-0432.CCR-05-2578DOI Listing
May 2006

c-Myc is able to sensitize human melanoma cells to diverse apoptotic triggers.

Melanoma Res 2004 Feb;14(1):3-12

Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.

Compared with other types of tumours, malignant melanomas are highly refractory to radio- or chemotherapy. To support the search for possible sensitizers, we explored the effects of the cellular oncoproteins c-Myc and N-Ras, which can decrease the clonogenic potential of irradiated p53-negative IGR39D melanoma cells. Using stable transfectants of this cell line, we showed that mutant N-Ras decreased the proliferation rate by inducing a prolonged cell cycle arrest. In contrast, c-Myc made these melanoma cells more prone to radiation-induced cell death. Membrane blebbing, the formation of apoptotic bodies and caspase activation, as measured by cleavage of Asp-Glu-Val-Asp (DEVD) substrate and poly(ADP-ribose) polymerase (PARP), indicate that these cells die by an apoptotic process. c-Myc also sensitized these p53-deficient melanoma cells to treatment with various cytotoxic drugs and heat shock. Similar results were obtained in inducible c-Myc models of IGR39D and in another melanoma cell line, 9007, which expresses functional p53. Together, these findings indicate that c-Myc is capable of sensitizing typically resistant tumour cells and that this occurs irrespective of the functional status of the p53 protein. Our results should facilitate the identification of factors that can be exploited for the treatment of aggressive cancers.
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http://dx.doi.org/10.1097/00008390-200402000-00002DOI Listing
February 2004

Analysis of T-cell responses in metastatic melanoma patients vaccinated with dendritic cells pulsed with tumor lysates.

Cancer Immunol Immunother 2004 Aug 3;53(8):715-22. Epub 2004 Mar 3.

Department of Clinical Oncology, Leiden University Medical Center, The Netherlands.

In melanoma patients, CD8+ cytotoxic T cells have been found recognizing self-proteins of which the expression is restricted to the melanocytic lineage. These melanocyte differentiation antigens are expressed in normal melanocytes as well as in 80-100% of primary and metastatic melanoma. In this report, six HLA-A*0201-subtyped metastatic melanoma patients vaccinated with dendritic cells (DCs) pulsed with autologous tumor lysates and keyhole limpet hemocyanin (KLH) were screened for the presence of CD8+ T cells specific for three HLA-A*0201-binding peptides derived from the melanosomal antigens MART-1/Melan-A, gp100, and tyrosinase. For this purpose, nonstimulated as well as in vitro peptide-stimulated peripheral blood mononuclear cells (PBMCs) were tested for peptide-specific IFN-gamma release by enzyme-linked immunosorbent spot (ELISpot) assays. Furthermore, expression of the melanosomal antigens MART-1/Melan-A, gp100, and tyrosinase in tumor lesions was analyzed by immunohistochemistry before and after vaccination. We also used the ELISpot technique to investigate whether KLH-specific T cells were induced and whether these cells released type 1 (IFN-gamma) and/or type 2 (IL-13) cytokines. Our data show induction of CD8+ T cells specific for the melanosomal peptides MART-1/Melan-A(27-35) or tyrosinase(1-9), as well as IFN-gamma-releasing KLH-specific T cells, in two of six vaccinated melanoma patients, but do not support an association between the induction of these T cells and clinical responses.
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http://dx.doi.org/10.1007/s00262-004-0514-zDOI Listing
August 2004

High intrinsic apoptosis, but not radiation-induced apoptosis, predicts better survival in rectal carcinoma patients.

Int J Radiat Oncol Biol Phys 2003 Oct;57(2):434-43

Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.

Purpose: An important feature of malignant tumors is the disturbance in the balance between proliferation and cell death. We evaluated the relevance of intrinsic and radiation-induced apoptosis and proliferation for prognosis in rectal cancer patients.

Methods And Materials: Patients were selected from a study that randomized for preoperative radiotherapy (RT). Apoptosis and proliferation were scored using specific antibodies in immunohistochemistry. The number of positive cells per square millimeter of carcinoma cells was determined in 98 randomly selected tumors, of which 45 had been irradiated. For the survival analyses, a cohort of 104 patients without positive circumferential resection margins was selected.

Results: In nonirradiated patients, high levels of intrinsic apoptosis correlated with better local control (p = 0.04) and better cancer-specific survival (p = 0.02). RT increased the median amount of apoptosis from 10.8 to 21.5 cells/mm(2) (p = 0.004), but this was not predictive for survival. The amount of proliferative cells was not altered after RT and had no influence on prognosis.

Conclusions: Intrinsic apoptosis correlated with both local control and cancer-specific survival, but proliferation was not predictive for prognosis. However, although RT increased apoptosis, its prognostic value was lost after RT. This is possibly because in rectal cancer, the proliferative status of tumors is always high and the aggressiveness of the tumor is determined by the number of "spontaneous" apoptotic tumor cells.
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http://dx.doi.org/10.1016/s0360-3016(03)00580-7DOI Listing
October 2003

CD4+ Th2 cell recognition of HLA-DR-restricted epitopes derived from CAMEL: a tumor antigen translated in an alternative open reading frame.

J Immunol 2003 Feb;170(3):1490-7

Department of Clinical Oncology, Leiden University Medical Center, The Netherlands.

Tumor Ag NY-ESO-1 is an attractive target for immunotherapy of cancer, since both CD8(+) CTL and CD4(+) Th cells against NY-ESO-1 have been described. Moreover, NY-ESO-1 as well as the highly homologous tumor Ag LAGE-1 are broadly expressed in various tumor types. Interestingly, the NY-ESO-1 and LAGE-1 genes also encode for proteins translated in an alternative open reading frame. These alternatively translated NY-ESO-ORF2 and CAMEL proteins, derived from the NY-ESO-1 and LAGE-1 genes, respectively, have been demonstrated to be immunogenic, since CTL specific for these proteins have been isolated from melanoma patients. In this study a panel of advanced melanoma patients was screened for the presence of Th cells specific for the alternatively translated tumor Ags NY-ESO-ORF2 and CAMEL. PBMC of melanoma patients were stimulated for 4 days with mixes of overlapping peptides covering the entire NY-ESO-ORF2 and CAMEL protein sequences and were tested for the release of type 1 (IFN-gamma) and type 2 (IL-13) cytokines in ELISPOT assays. In three of 15 patients, T cells specific for two CAMEL peptides (CAMEL(71-92) and CAMEL(81-102)) could be detected. From one of these patients, CD4(+) T cell clones specific for CAMEL(81-102) could be generated. These clones recognized a naturally processed epitope presented in both HLA-DR11 and HLA-DR12 and produced high levels of IL-4, IL-5, and IL-13. In conclusion, this study shows the presence of Th cells specific for the alternatively translated tumor Ag CAMEL in melanoma patients and is the first report that describes the isolation of tumor Ag-specific CD4(+) Th 2 clones.
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http://dx.doi.org/10.4049/jimmunol.170.3.1490DOI Listing
February 2003

Stretch-induced paracrine hypertrophic stimuli increase TGF-beta1 expression in cardiomyocytes.

Mol Cell Biochem 2002 Jul;236(1-2):147-53

Departments of Cardiology, Leiden University Medical Center, The Netherlands.

Cardiac hypertrophy refers to the abnormal growth of cardiomyocytes, and is often caused by valvular heart disease and hypertension. It involves the activation of growth, including increased protein synthesis and changes in gene expression. Transforming growth factor-beta1 (TGF-beta1) may play a central role in protecting the heart during the hypertrophic response by helping to restore normal functions of the affected myocardium. We tested the hypothesis that cardiomyocytes respond to stretch-induced paracrine hypertrophic stimuli with increased expression of TGF-beta1. To that purpose, we investigated whether angiotensin II (All), endothelin- I (ET-1) and TGF-beta, secreted by stretched cardiac and vascular cells, are involved in the paracrine mechanisms of stretch-induced changes of TGF-beta1 mRNA expression in stationary (i.e. non-stretched) cardiomyocytes. Our results indicated that TGF-beta1 mRNA expression in stationary cardiomyocytes was increased by AII release from cardiomyocytes that had been stretched for 30-60 min. Furthermore, it is likely that ET-1 and TGF-beta were released by stretched cardiac fibroblasts and endothelial cells to induce TGF-beta1 mRNA expression in stationary cardiomyocytes. Stretched vascular smooth muscle cells did not influence TGF-beta1 mRNA expression in stationary cardiomyocytes. These results indicate that AII, ET-I and TGF-beta, released by cardiac cell types, act as paracrine mediators of TGF-beta1 mRNA expression in cardiomyocytes. Therefore, we conclude that in stretched myocardium the cardiomyocytes, cardiac fibroblasts and endothelial cells take part in intercellular interactions contributing to cardiomyocyte hypertrophy.
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http://dx.doi.org/10.1023/a:1016138813353DOI Listing
July 2002

TCR reconstitution in Jurkat reporter cells facilitates the identification of novel tumor antigens by cDNA expression cloning.

Int J Cancer 2002 May;99(1):7-13

Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.

The identification of novel tumor antigens is of extreme importance for effective immunotherapy against cancer. A major obstacle in this field is the limited life span of tumor-specific cytotoxic T lymphocytes (CTLs) in vitro. Therefore we searched for a method to isolate the tumor specificity of these CTLs, i.e., their T-cell receptors (TCRs) and transfer it to an immortalized T-cell line. For this purpose, a TCR-negative Jurkat T-cell line was equipped with a nuclear factor of activated T cells (NFAT)-luciferase reporter construct to allow measurement of TCR-mediated activation. To establish the feasibility of this tumor-specific TCR transduction, we cloned the TCR genes of a known T-cell clone specific for the tumor antigen CAMEL (CTL-recognized antigen on melanoma) into a retroviral construct. Jurkat reporter cells transduced with this construct, Jrt-TCRalpha3beta5, were tested for their reactivity against CAMEL-expressing melanoma cells, peptide-loaded T2 cells and CAMEL-transfected COS-1 cells. The melanoma cell lines were poorly recognized, but peptide-pulsed and -transfected cells effectively stimulated NFAT signaling. The activation of TCR(+) Jurkat reporter cells was shown to be dependent on the antigen density on the target cells and the expression level of the coreceptor CD8 on the Jurkat cells. To verify the benefit of this TCR reconstitution method for identification of novel antigens, pools of the cDNA library from which CAMEL was originally cloned were transfected in COS-1 cells and screened with Jrt-TCRalpha3beta5. Identical cDNA pools were found that were positive with these cells and with the CAMEL-specific CTL clone. Our results illustrate that TCR-reconstituted Jurkat reporter cells are a useful tool in the identification of novel tumor antigens by cDNA expression cloning.
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http://dx.doi.org/10.1002/ijc.10317DOI Listing
May 2002

p53 expression in human rectal tissue after radiotherapy: upregulation in normal mucosa versus functional loss in rectal carcinomas.

Int J Radiat Oncol Biol Phys 2002 Mar;52(3):720-8

Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.

Purpose: In vitro, ionizing radiation of epithelial cells leads to upregulation of wild-type p53 and subsequent induction of p21(waf1). The effect of radiotherapy (RT) on the expression of these proteins in patients is unknown. We assessed the influence of RT on the expression of p53 and p21(waf1) in normal mucosa and rectal carcinomas in vivo.

Methods: Tumor and normal tissue samples were derived from rectal cancer patients randomized in a clinical trial in which the value of preoperative RT was evaluated. p53 and p21(waf1) expression was determined in 51 irradiated and 52 nonirradiated patients using immunohistochemistry.

Results: In normal mucosa, both p53 and p21(waf1) were strongly upregulated after RT compared with the expression in unirradiated normal tissue (p <0.001). In tumor cells, no significant difference in the expression of p53 or p21(waf1) was found in the irradiated vs. nonirradiated group. In the few rectal tumors with wild-type p53, induction of p53 after RT did not necessarily lead to upregulation of p21(waf1).

Conclusion: These findings demonstrate that in normal mucosa, a functional p53-p21(waf1) pathway is present, whereas in tumor cells it is defective in almost all cases because of either p53 mutation or down- or upstream disruption in tumors with wild-type p53. Therefore, we believe that the role of p53 expression as a single prognostic marker in rectal cancer needs reconsideration.
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http://dx.doi.org/10.1016/s0360-3016(01)02674-8DOI Listing
March 2002
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