Identification Of Tumor-specific Antigens Research Articles

EXTH-77. EXTENDED HALF-LIFE IL-2 SYNERGIZES WITH Α-PD-1 FOR TREATMENT OF PRECLINICAL MURINE GLIOBLASTOMA MODELS

Abstract α-PD-1 checkpoint blockade is a promising immunotherapeutic approach for many solid tumors; however, this strategy is minimally effective for GBM patients as a monotherapy. One potential reason why α-PD-1 checkpoint blockade is ineffective is the tumor microenvironment and systemic derangements of the immune system which are prevalent in GBM patients. We have successfully modeled peripheral immune derangements in experimental murine GBM models. These include low peripheral blood CD4 T cell counts, reduced MHC class II expression on monocytes, and atrophy of primary immune organs in animals harboring gliomas. We therefore hypothesized that extended half-life IL-2, a potent cytokine which promotes the proliferation, differentiation, and killing activity of T cells, could overcome immune derangements and could potentially synergize with α-PD-1 checkpoint blockade therapy. In cohorts of C57Bl/6 mice, we supplemented α-PD-1 checkpoint blockade with multiple bio-engineered extended half-life IL-2 molecules to treat GL261 and CT2A orthotopic glioma models which are normally resistant to monotherapy. We observe a statistically significant increase in survival in GL261-bearing animals receiving the combination of extended half-life IL-2 reagents and αPD-1 therapy relative to control-treated mice. Some animals receiving this combination therapy had a reduction in tumor burden to levels below detection. These long-term survivors underwent a rechallenge experiment with a second inoculation of GL261-Luc and effectively cleared tumor without further therapeutic intervention. This intervention causes a significant shift in the immune profile, including the formation of mature T cell responses, as well as restoring the diminished peripheral immune cell counts in GL261-bearing animals. Finally, combining extended half-life IL-2 with α-PD-1 checkpoint blockade is effective independently of the CD8 T cell response. Together, these data suggest that combination immunotherapies employing extended half-life IL-2 fusion proteins are translational to the clinic and do not require the identification of tumor specific antigens for efficacy

Biomaterial-Based In Situ Cancer Vaccines.

Cancer immunotherapies have reshaped the paradigm for cancer treatment over the past decade. Among them, therapeutic cancer vaccines that aim to modulate antigen-presenting cells and subsequent T cell priming processes are among the first FDA-approved cancer immunotherapies. However, despite showing benign safety profiles and the capability to generate antigen-specific humoral and cellular responses, cancer vaccines have been limited by the modest therapeutic efficacy, especially for immunologically cold solid tumors. One key challenge lies in the identification of tumor-specific antigens, which involves a costly and lengthy process of tumor cell isolation, DNA/RNA extraction, sequencing, mutation analysis, epitope prediction, peptide synthesis, and antigen screening. To address these issues, in situ cancer vaccines have been actively pursued to generate endogenous antigens directly from tumors and utilize the generated tumor antigens to elicit potent cytotoxic T lymphocyte (CTL) response. Biomaterials-based in situ cancer vaccines, in particular, have achieved significant progress by taking advantage of biomaterials that can synergize antigens and adjuvants, troubleshoot delivery issues, home, and manipulate immune cells in situ. This review will provide an overview of biomaterials-based in situ cancer vaccines, either living or artificial materials, under development or in the clinic, and discuss the design criteria for in situ cancer vaccines.

Retention time prediction boosts confidence in identification of tumorspecific antigens in HLA immunopeptidomics

Retention time prediction boosts confidence in identification of tumorspecific antigens in HLA immunopeptidomics

PODO447: a novel antibody to a tumor-restricted epitope on the cancer antigen podocalyxin

BackgroundThe success of new targeted cancer therapies has been dependent on the identification of tumor-specific antigens. Podocalyxin (Podxl) is upregulated on tumors with high metastatic index and its presence is...

Abstract B16: Identification of tumor-specific antigens shared by induced pluripotent stem cells

Abstract The development of cancer immunotherapeutics is limited by the tumor-specific antigen (TSA) discovery. Cancer cells and pluripotent stem cells, i.e., embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), show a significant transcriptomic overlap. Indeed, when used as prophylactic vaccines in another study, iPSCs prevented tumor growth in mouse models of lung, breast, and skin cancer, suggesting that we could mine iPSCs for the discovery of TSAs. We use a recently developed proteogenomic approach combining RNA-sequencing and mass spectrometry to characterize the immunopeptidome (MHC class I-associated peptides) of iPSCs. Analyses of hitherto one human iPS cell line allowed us to identify 9 MHC class I-associated antigens that were not expressed by normal somatic tissues and thus are likely immunogenic. 50% of these antigens were highly expressed in different cancer types from The Cancer Genome Atlas, highlighting their potential for cancer immunotherapy. Expression of these TSAs resulted from aberrant expression of noncoding genomic regions and not from mutations, making them ideal candidates for antigen-based cancer vaccines. Our study indicates which stem cell features present in cancer cells might be harnessed for cancer immunotherapy. Moreover, we show for the first time that the immunopeptidome of iPSCs reflects the state of pluripotency. Cancer stem cells are known to drive disease relapse after treatment, while cancer circulating cells possessing pluripotent stem cell features are more efficient at producing metastases. These factors indicate that treatments targeted at stemness could improve clinical outcome, and that iPSC-informed TSAs hold promise for prophylactic and therapeutic cancer vaccines. Citation Format: Anca Apavaloaei, Marie-Pierre Hardy, Jean-David Larouche, Qingchuan Zhao, Krystel Vincent, Jean-Philippe Laverdure, Chantal Durette, Joel Lanoix, Pierre Thibault, Claude Perreault. Identification of tumor-specific antigens shared by induced pluripotent stem cells [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2019 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(3 Suppl):Abstract nr B16.

Abstract B045: Antigen-free cancer vaccine to treat poorly immunogenic tumors

Abstract Certain chemotherapeutic drugs can elicit immunogenic death of tumor cells and enhance antitumor immune responses. Here we explore whether immunogenic chemotherapy can be utilized for the development of antigen-free cancer vaccines, by combining it with peritumorally injected biomaterial scaffolds that recruit dendritic cells (DCs) for subsequent antigen presentation and T-cell priming. Pore-forming alginate gels containing granulocyte-macrophage colony-stimulating factor (GM-CSF) and a doxorubicin-iRGD conjugate were found to efficiently induce the apoptosis of 4T1 triple-negative breast cancer cells in vivo, while recruiting large numbers of DCs. The co-encapsulation of CpG oligodeoxynucleotides in the gel significantly enhanced the immunogenic death of 4T1 cells, increased systemic tumor-specific CD8+ T-cells and tumoral infiltration of CD8+ T-cells, repolarized tumor-associated macrophages towards an inflammatory M1-like phenotype, and resulted in significantly improved antitumor efficacy. This in situ antigen-free gel vaccine shows promise for the treatment of poorly immunogenic tumors, and more broadly, may serve as a facile platform to enable in situ personalized cancer vaccination without requiring identification of tumor-specific antigens and manufacturing of personalized vaccines. Citation Format: Miguel C. Sobral, Hua Wang, Alexander J. Najibi, Aileen Li, Catia S. Verbeke, David J. Mooney. Antigen-free cancer vaccine to treat poorly immunogenic tumors [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B045.

Proteogenomic Platform for Identification of Tumor Specific Antigens.

Although immune responses to “cancer neoantigens” have been known for decades, the first neoantigen vaccines emerged only very recently. Current developments in genomics and proteomics have enabled descriptions of tumor mutational landscapes, and the immunogenicity of corresponding neoantigens can now be predicted either in silico or in vitro. Cancer regression could be achieved via a combination of neoantigen vaccination and an appropriate immunology approach. Research in model organisms and the results of initial clinical trials of neoantigen vaccines have shown them to be effective. We aim to emphasize the importance of neoantigen vaccines in personalized cancer treatment and describe their preparation. We summarize mutations leading to expression of an immunogenic antigen necessary for vaccine development. The processes leading to activation of T-cell anticancer immunity in a patient are briefly introduced. We especially focus on the identification of high confidence neoantigens by next-generation sequencing (NGS) and mass spectrometry (MS), which is key element in the process of designing neoantigen vaccines. Briefly, we describe a proteogenomic platform for confident identification of mutant peptides in biological material. We mention the possibility of neoantigen quantification in biological material using mass spectrometry such as SRM (selected reaction monitoring) and SWATH (sequential windowed acquisition of all theoretical fragment ion spectra). Successful clinical studies demonstrating the potential of neoantigen vaccination in personalized cancer treatment are summarized at the end of the paper. Key words: vaccination - mass spectrometry - neoplasms - organ-specific neoantigen This work was supported by the project MEYS - NPS I - LO1413. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Accepted: 9. 7. 2018.

OR10 Tumor antigens presented by the HLA-A*02:01 of breast cancer patient-derived xenografts

Aim Breast cancer is the second leading cause of cancer death in women, yet only a few antigens are available for directing immune therapies to a patient’s breast tumor. In this study we establish a new system for identifying tumor-specific antigens presented by the class I HLA of breast tumors. Our approach relies on generating a breast cancer HLA peptide-ligand reference database coupled with isolation of peptides from the HLA of patient-derived breast cancer xenografts (PDX). This HLA PDX approach can rapidly identify targets for existing and emerging cancer immune therapies. Methods A tumor-specific HLA reference peptide ligand library was constructed using 3 breast epithelial tumor cell lines and 1 healthy breast epithelial line. Cells were transfected to secrete HLA-A∗02:01, secreted HLA was affinity purified, HLA-bound peptides were eluted, separated by HPLC, and HLA-A∗02:01 breast cancer peptides were analyzed by mass spectroscopy. To validate the peptides as relevant targets for human breast tumors in vivo, HLA was extracted from PDX-derived tumors and the presence of specific peptides were confirmed in these primary tumors. Results In 20 PDX tumor lines, 7 of the tumors were A∗02:01, half of which expressed detectable A∗02:01. When evaluated for candidate peptide:HLA expression, the HCI-015 triple negative PDX xenograft expressed high levels of the Macrophage Inhibitory Factor 19–27 (MIF) peptide, a peptide that is expressed solely in breast cancer cell lines and primary tumors but not healthy tissues. We are currently expanding HCI-015 PDX for additional HLA extractions. Conclusions These data suggest that PDX tumors can be used as surrogates to human tumors for the direct identification of tumor-specific antigens. The use of a peptide library generated from several breast cancer cell lines allows peptides to be identified with increased sensitivity. Furthermore, expression of the candidate MIF 19-27 peptide on HCI-015 provides compelling evidence that certain tumor-antigens are conserved among cell lines, tumors, and in vivo models of breast cancer. Such antigens provide a unique therapeutic window that can be exploited for targeted immunotherapies.

The clinical-stage TLR5 agonist, Entolimod, mediates antimetastatic activity by stimulating a CXCR3-dependent NK-DC-CD8+ T cell axis

Abstract Activation of an anticancer innate immune response is an attractive immunotherapeutic opportunity that is challenged by insufficient safety of systemically administered innate immune modulators. Unusual tissue specificity of expression of TLR5 determines a uniquely safe profile of cytokines induced by the sole TLR5 agonist flagellin. Entolimod, a pharmacologically optimized flagellin derivative, was initially developed to treat and prevent acute radiation syndrome with demonstrated efficacy in rodents and non-human primates and safety in human healthy volunteers. In addition, Entolimod demonstrated antitumor effects in mouse models of uveal melanoma, lymphoma, breast, and colorectal carcinoma. Entolimod’s mechanism of action involves activation of NF-κB-, AP-1-, and STAT-3-driven immunomodulatory pathways in hepatocytes that initiates a cascade of cell-cell signaling events that mobilize innate and adaptive immunity to the liver. This includes CXCR3-dependent blood-borne NK cell homing followed by DC activation and antitumor CD8+ T cell memory formation. These results define systemically administered TLR5 agonists as an organ-specific immunoadjuvant enabling efficient antitumor vaccination that does not depend on identification of tumor-specific antigens. Therefore, Entolimod has strong promise as a safe, effective and broadly applicable immunotherapeutic agent against liver metastases, which are currently a major cause of cancer-associated mortality. Recent completion of a phase I trial of Entolimod in patients with advanced metastatic solid tumors has opened the opportunity to test whether efficacy demonstrated in animal cancer models could be translated into immunotherapy of human tumors.

Abstract 1294: Identification of tumor-specific antigens associated with RET/PTC3 expression

Abstract Relocated in transformation/papillary thyroid carcinoma, RET/PTC3 (RP3) is a fusion oncogene that causes a form of papillary thyroid cancer (PTC). In addition to driving transformation, the constitutively active kinase precociously phosphorylates itself as well as other intracellular proteins, thereby providing tumor-specific targets for the adaptive immune system. PTCs are known to have the ability to escape the immune system driving a dysregulation of the activity of several cell populations involved in the immune response. Indeed we found that mice immunized with RP3+/MHC class II+ cells produce dramatically fewer IFNg-secreting CD4+ T cells compared to mice immunized with RP3-/MHC class II+ counterparts as measured in the spleens. We demonstrated the immunogenicity of RP3 with reactivity in both BALB/c and C57BL/6 mice to RP3/IEd- and RP3/CIITA-expressing cells. Also, we observed immunogenicity of RP3-derived phosphopeptides in ELISpot assays, supporting the hypothesis that the aberrant autophosphorylation of RP3 is a source of tumor-specific immunogenicity. Finally, utilizing a RP3-expressing vaccinia vector for immunization of C57BL/6 mice, we identified peptide sequences that appear to be immunogenic on the basis of unique conformation of the fusion protein that impacts antigen processing. Taken together these results could pave the way for better vaccine approaches without accompanying autoimmunity. Citation Format: Laura Sponton, Gabriela Cosma, Mark Mendonca, Mirella Giovarelli, Laurence Crane Eisenlohr. Identification of tumor-specific antigens associated with RET/PTC3 expression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1294. doi:10.1158/1538-7445.AM2015-1294

Targeting the tumor microenvironment to enhance antitumor immune responses.

The identification of tumor-specific antigens and the immune responses directed against them has instigated the development of therapies to enhance antitumor immune responses. Most of these cancer immunotherapies are administered systemically rather than directly to tumors. Nonetheless, numerous studies have demonstrated that intratumoral therapy is an attractive approach, both for immunization and immunomodulation purposes. Injection, recruitment and/or activation of antigen-presenting cells in the tumor nest have been extensively studied as strategies to cross-prime immune responses. Moreover, delivery of stimulatory cytokines, blockade of inhibitory cytokines and immune checkpoint blockade have been explored to restore immunological fitness at the tumor site. These tumor-targeted therapies have the potential to induce systemic immunity without the toxicity that is often associated with systemic treatments. We review the most promising intratumoral immunotherapies, how these affect systemic antitumor immunity such that disseminated tumor cells are eliminated, and which approaches have been proven successful in animal models and patients.

Targets for active immunotherapy against pediatric solid tumors.

The potential role of antibodies and T lymphocytes in the eradication of cancer has been demonstrated in numerous animal models and clinical trials. In the last decennia new strategies have been developed for the use of tumor-specific T cells and antibodies in cancer therapy. Effective anti-tumor immunotherapy requires the identification of suitable target antigens. The expression of tumor-specific antigens has been extensively studied for most types of adult tumors. Pediatric patients should be excellent candidates for immunotherapy since their immune system is more potent and flexible as compared to that of adults. So far, these patients do not benefit enough from the progresses in cancer immunotherapy, and one of the reasons is the paucity of tumor-specific antigens identified on pediatric tumors. In this review we discuss the current status of cancer immunotherapy in children, focusing on the identification of tumor-specific antigens on pediatric solid tumors.

Targeting epidermal growth factor receptor variant III: a novel strategy for the therapy of malignant glioma

Immunotherapy represents an exciting approach for the treatment of glioblastoma multiforme. The success of this approach depends on the identification of tumor-specific antigens that can then be exploited to target the tumor. One such antigen, the epidermal growth factor receptor variant III (EGFRvIII) is found in up to 50% of malignant gliomas. This molecule is an interesting vaccine candidate since its expression is tumor specific. Recent preclinical, as well as clinical, studies of EGFRvIII-based vaccines suggest a promising role for immunotherapy of malignant gliomas. This review focuses on vaccines that target EGFRvIII. We review preclinical and clinical data and emphasize the immunological mechanisms involved in its therapeutic effect.

Potential target antigens for immunotherapy in human pancreatic cancer

Background To be effective and selective, immunotherapy ideally targets specifically tumor cells and spares normal tissues. Identification of tumor specific antigens is a prerequisite to establish an effective immunotherapy. Still very little is known about the expression of tumor-related antigens in pancreatic neoplasms. Cancer Testis antigens (CT) are antigens shared by a variety of malignant tumors, but not by normal tissues with the exception of germ cells in testis. Restricted expression in neoplastic tissues and inherent immunogenic features make CT antigens ideal for use in immunotherapy. We analyzed the expression of a selected panel of nine CT antigens that have been proven to elicit an efficient immunogenic response in other malignancies. In addition we analyzed the expression of HERV-K-MEL, an immunogenic antigen of viral origin. Methods Pancreatic adenocarcinoma tumor samples ( n = 130) were obtained intraoperatively, control tissues ( n = 23) were collected from cadaveric donor and from patients with chronic pancreatitis. Tumor-associated antigen expression of MAGE-A1, MAGE-A3, MAGE-A4, MAGE-A10, LAGE-1, NY-ESO-1, SCP-1, SSX-2, SSX-4 and HERV-K-MEL was assessed by PCR. Sequencing of PCR products were performed to assess the expression of SSX-4 in neoplastic and normal pancreatic tissues. Results Three of 10 tested antigens were expressed in over 10% of malignant pancreatic tissue samples. SSX-4 was found positive in 30% of cases, SCP-1 in 19% and HERV-K-MEL in 23% of cases. No expression of CT antigens was found in non-malignant pancreatic tissue with the exception of SSX-4 and and SSX-2. Conclusions Fifty two percentage of the analyzed tissues expressed at least one CT antigen. The concomitant expression of SSX-4 in both malignant and non-malignant pancreatic tissue is a new finding which may raise concerns for immunotherapy. However, HERV-K-MEL is expressed with a relatively high prevalence and may be a candidate for specific immunotherapy in a large subgroup of pancreatic cancer patients. This study advocates the analysis of patients with regard to their immunogenic profile before the onset of antigen-specific immunotherapy.

SELECTIVE EXPRESSION OF TUMOR ASSOCIATED ANTIGENS BY SAMPLES DERIVED FROM HUMAN PANCREATIC CANCER

Background: To be effective and selective, immunotherapy ideally targets specifically tumor cells and spares normal tissues. Identification of tumor specific antigens is a prerequisite to establish an effective immunotherapy. Still very little is known about the expression of tumor-related antigens in pancreatic neoplasms. Cancer Testis antigens (CT) are antigens shared by a variety of malignant tumors, but not by normal tissues with the exception of germ cells in testis. Restricted expression in neoplastic tissues and inherent immunogenic features make CT antigens ideal for use in immunotherapy. We analysed the expression of a selected panel of ten CT antigens that have been proven to elicit an efficient immunogenic response in other malignancies. In addition we analyzed the expression of HERV-K-MEL, an immunogenic antigen of viral origin. Methods: Pancreatic adenocarcinoma tumor samples (n = 117) were obtained intraoperatively, control tissues (n = 5) were collected from cadaveric donor and from patients with chronic pancreatitis. Tumor-associated antigen expression of MAGE-A1, MAGE-A3, MAGE-A4, MAGE-A10, LAGE-1, NA-17A, NY-ESO-1, PRAME, SCP-1, SSX-2, SSX-4 and HERV-K-MEL was assessed by PCR. Sequencing of PCR products were performed to assess the expression of SSX-4 in neoplastic and normal pancreatic tissues. Results: SSX-4 was found positive in 33 % of cases, SCP-1 in 16% and HERV-K-MEL in 22% of cases. No expression of CT antigens was found in healthy pancreatic tissue with the exception of SSX-4 and, to a lesser extent, SSX-2. Conclusions: 51% of tissues analyzed expressed at least one CT antigen. The concomittant expression of SSX-4 in both malignant and normal pancreatic tissue is a new finding which may raise concerns for immunotherapy. However, HERV-K-MEL is expressed with a relatively high prevalence and may be a candidate for specific immunotherapy in a large subgroup of pancreatic cancer patients. This study advocates the analysis of patients with regard to their immunogenic profile before the onset of antigen-specific immunotherapy.

Protein Kinase C Blockade Inhibits Differentiation of Myeloid Blasts into Dendritic Cells by Calcium Ionophore A23187

Direct differentiation of myeloid leukemia blasts into antigen-presenting dendritic cells (DCs) for use as cellular vaccines is unique in that identification of tumor-specific antigens may not be necessary because the antigens should already be endogenously expressed. We hypothesized that signaling through protein kinase C (PKC) is required for differentiation of HL-60 promyeloblasts into DCs upon stimulation with calcium ionophore A23187. To demonstrate the inhibitory effect of PKC blockade, we pretreated HL-60 myeloid blasts with the protein kinase inhibitor bisindolylmaleimide I (Bis-1) for 24 hours and then treated the cells with calcium ionophore A23187 for an additional 24 hours. Controls consisted of HL-60 blasts treated with A23187, Bis-1 alone, or media. We noted that blasts cultured in media, Bis-1, or Bis-1 then A23187 did not develop the morphologic and phenotypic DC characteristics, up-regulate Rel B, or activate allogeneic T-cells. Our findings suggested that PKC blockade inhibits morphologic, phenotypic, and functional differentiation of HL-60 promyeloblasts into antigen-presenting DCs. Our findings supported the role of PKC as an obligatory pathway for calcium ionophore A23187-induced differentiation of HL-60 myeloblasts into antigen-presenting DCs.

Functional characterization of hepatoma-specific stem cell antigen-2.

Identification of tumor-specific antigens and genetic pathways may lead to potential diagnostic and therapeutic applications in cancer treatment. cDNA microarray has been used in cancer gene profiling, but the broad spectrum of data accruing and narrow signal-to-noise range of this technology have limited its use in rapid identification of highly differentially expressed tumor genes. Here, we used a modified suppression subtractive hybridization (SSH) method to isolate a small number of highly differentially expressed genes from murine hepatoma cells. For functional analysis of these hepatoma-specific genes, we employed the small interference RNA (siRNA)-mediated gene silencing method with lentiviral vectors, which have the advantages of high delivery efficiency and long lasting effect. Stem cell antigen-2 (Sca-2) was identified as one of the highest differentially expressed tumor antigens. Lentiviral siRNA successfully suppressed >90% of Sca-2 expression and the suppression lasted longer than 3 mo. Interestingly, inhibition of Sca-2 induced rapid hepatoma cell apoptosis, and the survival Sca-2-negative hepatoma cells exhibited high sensitivity to extrinsic tumor necrosis factor alpha (TNF-alpha) apoptosis signal but not intrinsic apoptosis signal. Analysis of TNF receptor 1 (TNFR1) by flow cytometry and Western blotting indicated that Sca-2 expression downregulated cell surface but not de novo synthesis of TNFR1 in the hepatoma cells. Together, our results suggested that Sca-2 was a signal transducer situated at the nexus of surface molecules regulating death receptor-mediated apoptosis. The technology illustrated that this method can deduce a small number of highly differentially expressed tumor genes that may have diagnostic and therapeutic potential.

P-473 Identification of tumor-specific antigens recognized by antibodies derived from tumor-infiltrating B lymphocytes in lung cancer patients

P-473 Identification of tumor-specific antigens recognized by antibodies derived from tumor-infiltrating B lymphocytes in lung cancer patients

Immunité anti-tumorale et thérapies cellulaires du cancer

The identification of tumor specific antigens has provided important advance in tumor immunology. It is now established that specific cytotoxic T lymphocytes (CTL) and natural killer cells infiltrate tumor tissues and are effector cells able to control tumor growth. However, such a natural antitumor immunity has limited effects in cancer patients. Failure of host defenses against tumor is consecutive to several mechanisms which are becoming targets to design new immunotherapeutic approaches. CTL are critical components of the immune response to human tumors and induction of strong CTL responses is the goal of most current vaccine strategies. Effectiveness of cytokine therapy, cancer vaccines and injection of cells improving cellular immunity have been established in tumor grafted murine models. Clinical trials are underway. To day, interest is particularly focused on cell therapy: injected cells are either "ready to use" effector cells (lymphocytes) or antigen presenting cells able to induce a protective immune reaction in vivo (dendritic cells). The challenge ahead lie in the careful optimization of the most promising strategies in clinical situation.

Serological detection of cutaneous T-cell lymphoma-associated antigens

Cutaneous T-cell lymphomas (CTCL) are a group of skin neoplasms that originate from T lymphocytes and are difficult to treat in advanced stages. The present study is aimed at the identification of tumor-specific antigens from a human testis cDNA library using human sera known as the SEREX (serological identification of recombinantly expressed genes) approach. A cDNA library from normal testicle tissue was prepared and approximately 2 million recombinants were screened with sera from Sézary Syndrome and Mycosis fungoides patients. A total of 28 positive clones belonging to 15 different genes/ORFs were identified, including five hitherto unknown sequences. Whereas control sera did not react with most clones, 11-71% sera from CTCL patients were reactive against the identified clones. Expression analysis on 28 normal control and 17 CTCL tissues by reverse transcription-PCR (RT-PCR) and Northern blotting revealed seven ubiquitously distributed antigens, six differentially expressed antigens (several normal tissues were positive), and two tumor-specific antigens that were expressed only in testis and tumor tissues: (i) A SCP-1-like sequence, which has already been detected in various tumors, has been found in one CTCL tumor and four sera of CTCL patients reacted with various SCP-1-like clones and (ii) a new sequence named cTAGE-1 (CTCL-associated antigen 1) was detected in 35% of CTCL tumor tissues and sera of 6/18 patients reacted with this clone. The present study unravels CTCL-associated antigens independent of the T-cell receptor. The SCP-1-like gene and cTAGE-1 were shown to be immunogenic and immunologically tumor-specific and may therefore be candidates for immunotherapy targeting CTCL.