Expression of serologically identified tumor antigens in acute leukemias

doi:10.1016/S0145-2126(02)00230-8

Leukemia Research

Volume 27, Issue 7, July 2003, Pages 655-660

https://doi.org/10.1016/S0145-2126(02)00230-8 Get rights and content

Abstract

Cancer/testis antigens (CTA) are an expanding family of immunogenic proteins selectively expressed in human neoplasms. As little is known about the expression of serologically identified CTA in leukemias so far, we investigated the expression of 5 CT genes (SSX-1, HOM-MEL-40/SSX-2, HOM-TES-14/SCP-1, SCP-3 and NY-ESO-1) in leukemic blood samples obtained from patients with either acute lymphatic leukemias (ALL) or myelocytic leukemia (AML). RT-PCR-analyses showed no expression of any of the CT-genes in the leukemia samples of 19 patients with AML, whereas frequent expression was found in ALL. In the 17 ALL cases studied, SCP3a, SSX-1, HOM-MEL-40/SXX-2 and HOM-TES-14/SCP-1 were expressed in 47, 29, 29 and 12%, respectively, whereas no case was positive for NY-ESO-1. 65% of patients with ALL showed expression of at least one, 41% of two or more of the five CT-genes investigated. We conclude that a majority of the ALLs might be amenable for specific immunotherapeutic interventions. However, the identification of additional antigens with a frequent expression in leukemias is warranted to allow the development of widely applicable polyvalent leukemia vaccines.

Introduction

Although a variety of therapeutic strategies, such as allogeneic and autologous bone marrow transplantation combined with high-dose chemotherapy, have been developed, only approximately 30–40% of adult patients with acute lymphatic leukemias (ALL) are cured. In between the different types of ALLs, the range of the percentage of cured patients varies from 0% for certain bcr/abl-positive subtypes to 60–70% for patients with B-ALL [1], [2]. As in solid tumors (e.g. malignant melanoma) antigen specific immunotherapies are being pursued with promising results [3], [4], similar innovative strategies are required for leukemia.

However, a prerequisite for the implementation of such specific therapeutic strategies is the identification and availability of molecularly defined antigens that are either exclusively or preferentially expressed in malignant tissue. A new and expanding class of tumor antigens, known as cancer/testis (CT) antigens, has recently been identified by expression cloning using spontaneous immune reponses from cancer patients as screening probes [5]. CT genes are testis-specific genes, which are normally silenced in adult somatic tissues but activated aberrantly in various cancer cells [6]. Members of the MAGE [7], BAGE [8] and GAGE family [9] identified by tumor cell reactive T lymphocytes as well as antigens identified serologically using the SEREX-approach [10], [11] including SSX family members [12], [13], [14] NY-Eso-1 [15] and HOM-Tes-14/SCP-1 [16] belong to this class. The tumor restricted expression pattern of these antigens makes them to interesting targets for immunotherapeutic approaches.

Whereas the expression of MAGE antigens in leukemia has been previously described, only limited information is available about the expression of serologically identified CT antigens in acute leukemia. We therefore investigated the composite expression of several serologically defined CT genes in acute leukemia including AML and ALL.

Section snippets

Patient material

This study had been approved by the local ethical review board (“Ethikkommission der Ärztekammer des Saarlandes”). Recombinant DNA work was done with the official permission and in accordance with the rules of the state government of Saarland. Patients that had been diagnosed at the Departments of Hematology and Pediatrics, University of Saarland according to routine clinical standards. French–American–British-classification were used for the subtype determination of the AML samples. Patients

Study population and validity of the experimental approach

In total, 36 leukemia specimens (17 ALL, 19 AML) were investigated for the expression of the five CT genes SSX-1, HOM-MEL-40/SSX-2, HOM-Tes-14/SCP-1, SCP-3a and NY-ESO-1. Only cDNA preparations that had been assessed for cDNA integrity by amplification of an 800 bp β-actin product were included into this study. To exclude false positive results due to small amounts of contaminating genomic DNA in the RNA preparation, the individual primer sets were chosen to correspond to sequences located in

Discussion

We report that a high percentage of patients with acute lymphatic leukemia express antigens encoded by members of the CT gene family. The frequency of expression of individual CT genes ranged from 0% for NY-ESO-1 to 47% for SCP-3a. Nearly 2/3 of the specimens obtained from ALL patients express at least one of the five CT genes investigated in this study. In contrast, we detected no expression of any of the CT genes in neoplastic cells obtained from AML patients. Interestingly, the same type of

Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (SA 776/2-1 to US). Conception and design by Ö. Türeci, M. Pfreundschuh and U. Sahin; analysis and interpretation of data by P. Niemeyer, Ö. Türeci, T. Eberle, M. Pfreundschuh and U. Sahin; drafting the article by P. Niemeyer; critical revision of the article for important intellectual content by Ö. Türeci, N. Graf, M. Pfreundschuh and U. Sahin; final approval by P. Niemeyer, M. Pfreundschuh and U. Sahin; provision of study

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Consistently, SCP3 is preferentially expressed in multiple cancer cells. For instance, human acute lymphatic leukemias show frequent expression of SCP3 (47%) [9]. Recently, we also have demonstrated that SCP3 is expressed in human cervical cancer cells [10].
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Promoter methylation has been established as one important factor regulating the expression of CTA genes [17], and several studies showed that in vitro treatment with DNA demethylating agents results in an enhanced expression of CTAs in tumor cell lines [18–22]. In contrast to solid tumors, prototypical members of the CTA family such as NY-ESO-1 or MAGEA1 are not expressed in myeloid leukemias [23–25]. One study explored the effect of the demethylating agent DAC on CTA expression in primary leukemic cells in vivo[26].
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Due to the very restricted expression pattern of CT antigens we considered it very unlikely that the strong and commonly observed antibody responses in our patients were simply part of a general autoimmune phenomenon in patients who had undergone alloSCT. Nevertheless, we analyzed post-alloSCT sera from 40 patients with acute myeloid leukemia (AML), which is considered an entity lacking expression of most CT antigens,20,27,28 for CT antigen–specific antibodies. All 40 patients with AML had experienced full engraftment and complete immune reconstitution at the time the sera were collected.
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Expression of serologically identified tumor antigens in acute leukemias

Abstract

Introduction

Section snippets

Patient material

Study population and validity of the experimental approach

Discussion

Acknowledgements

Immunity

Akute lymphatische Leukämie des Erwachsenen

Internist

Treatment of adult acute lymphoblastic leukemia

Schweiz Rundsch Med Prax

Tumor regressions observed in patients with metastatic melanoma treated with an antigenic peptide encoded by gene MAGE-3 and presented by HLA-A1

Int J Cancer

Identification of NY-ESO-1 Epitopes presented by human histocompatibility Antigen (HLA)-DRB4*0101-0103 and recognized by CD4+ T Lymphocytes of patients with NY-ESO-1 expressing melanoma

J. Exp. Med.

New paths in human cancer serology

J Exp Med

Cancer-testis antigens, targets for cancer immunotherapy

Cancer J Sci Am

A gene encoding an antigen recognized by cytolytic T-lymphocytes on a human melanoma

Science

A new family of genes coding for an antigen recognized by autologous cytolytic T lymphocytes on a human melanoma

J. Exp. Med.

Human neoplasms elicit multiple immune response in the autologous host

Proc. Natl. Acad. Sci. USA

Serological analysis of human tumor antigens: molecular definition and implications

Molec. Med. Today

The SSX-2 gene, which is involved in the t (X;18) translocation of synovial sarcomas, codes for the human tumor antigen HOM-MEL-40

Cancer Res.

Expression of SSX genes in human tumors

Int. J. Cancer

SSX: a multigene family with several members transcribed in normal testis and human cancer

Int. J. Cancer