NY-ESO-1 Research Articles

Relationship between circulating tumor-associated autoantibodies and clinical outcomes in advanced-stage NSCLC patients receiving PD-1/−L1 directed immune checkpoint inhibition.

BackgroundDurable tumor regressions are observed in a subset of advanced-stage non-small cell lung cancer (NSCLC) patients receiving PD-1/−L1 targeted immune checkpoint inhibitors (or ‘immunotherapy’) alone or in combination with chemotherapy. However, the majority of advanced NSCLC patients receiving these agents do not experience long-term disease control. Existing methods to identify patients most likely to gain clinical benefit from PD-1/−L1 immunotherapy have limitations, creating a need for improved methods to guide treatment selection, particularly for those likely to benefit from single-agent immunotherapy. Here, we describe the development of a series of novel assays for tumor-associated autoantibodies as part of an exploratory study intended to determine if these biomarkers have potential prognostic value in this setting. MethodA selection of recombinant tumor autoantigens previously characterized for their diagnostic utility were developed and preliminarily evaluated by this study. These include: Fumarate Dehydrogenase (FH), Hydroxysteroid 17-Beta Dehydrogenase 10 (HSD17B10), Inosine Monophosphate Dehydrogenase 2 (IMPDH2), New York Esophageal Squamous Cell Carcinoma-1 (NY ESO-1), Phosphoglycerate Mutase 1 (PGAM1), and Vimentin. Custom Luminex immunobead assays were developed for these targets to quantitatively assess autoantibody levels in individual patient sera. Assays were erected as indirect immunoassays on MagPlex® Microspheres using standard carbodiimide/NHS-based chemistries, utilizing a biotin-conjugated secondary (i.e. anti-human IgG) antibody and R-phycoerythrin-conjugated streptavidin reporter system. Standard curves were created for quantitative purposes using commercially-available anti-antigen antibodies and permitted analytical performance characteristics to be calculated. These assays were used to preliminarily evaluate a series of pretreatment serum samples from stage IV NSCLC patients receiving anti PD-1/−L1 therapy after failure of at least one prior line of therapy (n = 40) and their classification efficiency calculated based on 12 months overall survival (OS) threshold. ResultsSix assays were developed that each showed dynamic ranges of four orders of magnitude and provided more than 90% classification accuracy based on the observed clinical outcome data. Inter- and intra-assay precision was assessed within these standards and overall %CVs of ≤7% and ≤ 10%, respectively, were calculated. Generally, the baseline level of autoantibodies were significantly (p < 0.05) lower in the ≥12 months survival group relative to the <12 months survival groups. Serum titers of FH, HSD170B, NY-ESO-1, and vimentin were significantly correlated with ≥12 month survival (p-value 0.0038, 0.0061, 0.0073, and 0.022, respectively). IMPDH2 and PGAM1 were found to have marginal significance (p-value 0.08 and 0.076, respectively). ConclusionThis study demonstrates an efficient and promising means for assessing circulating autoantibody titers that could be useful in selecting advanced NSCLC patients for PD-1/−L1 directed immunotherapy. Further exploration and validation of this paradigm is warranted to further refine current treatment selection methods for this therapeutic strategy.

Overexpression of NKG2D-CD3ζ in NY-ESO-1 TCR-T cells enhanced cytotoxicity to acute myeloid leukemia cells in vitro

Overexpression of NKG2D-CD3ζ in NY-ESO-1 TCR-T cells enhanced cytotoxicity to acute myeloid leukemia cells in vitro

Abstract 426: Autoantibodies: A promising prognostic tool for immunotherapy response in advanced non-small cell lung cancer

Abstract Background: Immune-checkpoint blockade has revolutionized cancer therapy in advanced non-small cell lung cancer (NSCLC). Tissue expression of programmed death protein ligand (PD-L1) remains the gold standard for patient stratification, however, the limited performance of this marker encourages investigations for improved molecular diagnostics. The objective of this study is to identify and evaluate the role of neoantigen-associated autoantibodies to predict the clinical response to anti-PD-1/-L1 in advanced stage NSCLC. Method: Lung adenocarcinoma A549 and H358 cell lysate proteins were resolved via 2-dimensional electrophoresis, electroblotted onto nitrocellulose, and immunoprobed with pooled, pretreatment sera (n= 4/ group) derived from patients with advanced NSCLC who received PD-1/-L1 directed immunotherapy. These patients have documented disease progression within 12 weeks (“rapid progression”) or demonstrated radiographical stable disease/progression after the first 180 days of therapy (“late progression”). Immunoreactive spots were detected with an HRP-conjugated, anti-human IgG secondary antibody with digital densitometry. A 4-fold cutoff threshold in expression was used to prioritize spots for identification via tandem mass spectrometry. From A549 cells, recombinant proteins were selected for STIP-1, annexin A2, HSPA8, and GAPDH. These proteins were then analyzed via immunoblotting methods using sera from each indicated group (n=4 per group). In addition, identified proteins from H358 cells include FH, HSP70B, IMPDH2, NY ESO-1, PGAM-1, and vimentin. Recombinant versions of a selection of autoantigens identified in this manner were commercially acquired and used to develop custom Luminex immunobead assays to quantitatively assess autoantibody production in individual patient sera (rapid progressors, n=14; late progressors, n=18). Values were statistically compared via Mann-Whitney test. Results: Series of differentially expressed autoantigens predictive of clinical response to PD-1/-L1 directed immunotherapy were identified. Western blots of neoantigens identified from A549 cells; STIP-1, annexin A2, HSPA8, and GAPDH were significantly able to distinguish between response groups (p-value &amp;lt; 0.001). Six H358 targets resulting from the custom bead-based immunoassay development; FH, HSP70B, IMPDH2, NY-ESO-1, PGAM-1, and vimentin were also able to distinguish between groups (p-values of 0.01, 0.01, 0.022, 0.005, 0.034, and 0.027, respectively). Conclusion: Our study demonstrates that serum autoantibodies have great promise to serve as a robust tool to prognosticate response for patients receiving PD-1/-L1 directed immunotherapy and potentially aid current treatment selection methods. Additional targets are currently being developed into multiplexed immunobead assays for evaluation across larger cohorts of patients. Citation Format: Imad Tarhoni, Cristina Fhied, Melissa Pergande, Revathi Kollipara, Connor J. Wakefield, Katherine Gallo, Apoorva Tangri, Marta Batus, Mary Jo Fidler, Philip Bonomi, Jeffrey A. Borgia. Autoantibodies: A promising prognostic tool for immunotherapy response in advanced non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 426.

Delivery of Cancer-Testis Antigens Using Self-Assembled Dipeptide Nanotubes Inhibits Tumor Growth in Mice Melanoma Model

Delivery of tumor-associated antigens for cancer immunotherapy is generally limited due to poor stability and low cellular uptake. Use of nanoparticles as delivery vehicles has been an exciting development but toxicity associated with nanoparticles has remained a concern. Here, we have used conformationally constraint self-assembling dipeptide nanotubes for the delivery of Cancer-Testis antigens (CTAs). A dipeptide Phenylalanine-α, β-dehydrophenylalanine (F-ΔF) was synthesized and was found to self-assemble into nanotubes. CTAs (NY-ESO-1 and MAGE-3) were also synthesized and loaded onto F-ΔF nanotubes using physical adsorption method for delivery purposes. CTA loaded F-ΔF nanotubes showed higher cellular uptake in macrophages. In animal studies, F-ΔF-CTA nanocomplex elicited much higher immune response compared to CTAs alone and showed an inhibition of ~65-70% tumor growth in mouse melanoma model. Our results indicated that F-ΔF nanotubes are biocompatible and are efficient delivery vehicles for CTAs in vivo and therefore are excellent candidates for further development as delivery agents.

Correlation between the high expression levels of cancer-germline genes with clinical characteristics in esophageal squamous cell carcinoma.

Antigens encoded by cancer-germline genes are attractive targets for cancer immunotherapy. In this study, we aimed to evaluate the mRNA expression of cancer-germline genes, expression of the encoded proteins in patients with esophageal squamous cell carcinoma (ESCC) and their correlations with clinical characteristics. In addition, the effects of downregulation cancer-germline genes on ESCC cells were assessed in vitro. Our results showed that cancer-germline genes were frequently expressed in ESCC samples. The positive rates of in ESCC samples were: 87% of MAGE-A3, 60% of MAGE-A4, 65% of MAGE-C2, and 20% of NY-ESO-1 at mRNA level. MAGE-A3 expression was associated with age, lymph node metastasis and tumor stage (all P<0.05), while MAGE-C2 expression was only associated with tumor stage (P<0.05). Furthermore, the MAGE-A3 expressing patients had a poorer overall survival (P<0.05). Multivariate analysis identified MAGE-A3 as an independent poor prognostic marker in ESCC. In vitro assay, ESCC cell lines treated with specific siRNAs to down-regulate MAGE-A3 and MAGE-C2 resulted in decreased colony-formation and migration ability (P<0.05). Epithelial marker E-cadherin was up-regulated in siRNA-MAGE-A3/C2 cells compared to controls, whereas mesenchymal markers Vimentin, N-cadherin and Slug were downregulated (all P<0.05), suggesting a role for MAGE-A3/C2 in ESCC metastasis through inducing epithelial-mesenchymal transition. The present study revealed that cancer-germline genes and their encoded proteins were frequently expressed in ESCC tumor samples and were related to poor prognosis. Thus, cancer-germline genes may serve as useful biomarkers and potential targets for ESCC patients.

Construction of pRV-Display-NY-ESO-1 eukaryotic plasmid and its stable transfected Myc-CaP cell line

Objective To construct the pRV-Display-NY-ESO-1 eukaryotic plasmid,and establish Myc-CaP cell line transfected with NY-ESO-1.Methods The full sequence of NY-ESO-1 was get from Genbank and amplified by polymerase chain reaction (PCR).Insert the fragments into p-Display eukaryotic expression vector.The recombinant plasmid with targeted gene was identified by digestion.Display-NY-ESO-1 was amplified by PCR and inserted into pRV retroviral vector.Retrovirus was produced by Phoenix cell line,and transfected into Myc-CaP cell line.Results The recombinant plasmid pRV-Display-NY-ESO-1 was successfully constructed and identified by PCR and DNA sequencing.Green fluorescence was observed on Phoenix cells and Myc-CaP cells encoding with GFP.The purity of Myc-CaP cells encoded with NY-ESO-1 was more than 80％ according to flow cytometry.Conclusion Myc-CaP cell line was stably transfected with pRV-Display-NY-ESO-1,which is the base for further experiments. Key words: NY-ESO-1 ; Myc-CaP cell line; Recombinant plasmid; Construction

The killing effect of NY-ESO-1-sensitzed dendritic cells-induced cellular toxic lymphocyte on human retinoblastoma cell

Background Cell immunologic therapy for retinoblastoma （RB）is becoming a hot research topic. Cancer-testis antigen is a human immunogenic protein and is used to treat some tumors. However,its effect on RB has not been investigated. Objective The present study was to discuss the antigen specific anti-tumor effect of cytotoxic T lymphocytes （CTL）induced by the cancer-testis antigen, NY-ESO-l-sensitized dendritic cells （DCs）, on human RB. Methods PCR was performed to amplify target gene fragments from the NY-ESO-1 plasmids,and then the target gene fragments were digested with the restriction enzymes SalI and EcoRI. Harvested fragments were inserted into the pDC316 plasmid to construct the recombinant plasmid pDC316/NY-ESO-1. The expression of NY-ESO-1 protein in human RB cells strain, HXO-RB44,was detected by immunofluoreseence and Western blot. Monoeytes wereisolated from 60 ml of peripheral blood from a healthy donor using Ficoll density-gradient centrifugation with a cell density of 1 x 107/ml. DCs isolated from blood were stimulated with recombinant human granulocyte-macrophage colony stimulating factor （rhGM-CSF） and recombinant human interleukin-4 （ rhIL-4 ）. The recombinant plasmid pDC316/NY-ESO-1 was transfected into DCs and the DCs were co-cultured with T lymphocytes. The resultant CTL were used as effector ceils. The growth of the CTL was detected by MTT assay. The CTL were then added into the growth medium used for culturing HXO-RB44 cells and the vitality of the HXO-RB44 cells was assayed by MTT assay. Results The sequence of the cloned DNA fragment of the recombinant plasmid pDC316/NY-ESO-lwas conforms with the sequence of the NY-ESO-1 gene. The expression of the NY-ESO-1 protein in HXO-RB44 cells was tested by immunofluorescence and Western blot. DCs were successfully induced with rhIL-4 and rhGM-CSF from PBMC. The recombinant expression plasmid pDC316/NY-ESO-1 was successfully transferred into DCs. These DCs had high expression of surface molecules such as HLA-DR （42. 1% ） , CD80 （54.2%） , CD83 （39.7%） and CD86 （94.8%）. The CTL that was induced by DCs-sensitized with NY-ESO-1 specifically killed HXO-RB44 cells. CTL induced by the sensitized DCs had a stronger cytotoxic effect against HXO-RB44 cells compared with un-sensitized DCs and CTL un-induced with DCs, as shown by MTT asssay（ P〈0.05 ）. The anti-tumor activity was highest when the ratio of effector to target was 75 ： 1 （ P〈0. 05 ）. Conclusions DCs transfected by the recombinant plasmid pDC316/ NY-ESO-1 can induce the proliferation of allogenic CTLs, which showed a specific anti-tunmr effect against HXORB44 cells. These results present a new type of immunotherapy for the treatment of RB. Key words: Retinoblastoma; Ddendritic cell; Cytotoxic T lymphocyte; NY-ESO-1

NY-ESO-1 expression in synovial sarcoma and other mesenchymal tumors: significance for NY-ESO-1-based targeted therapy and differential diagnosis

AbstractA promising targeted therapy against NY-ESO-1 (CTAG 1B) using genetically modified T-cells in synovial sarcomas was recently demonstrated in a clinical trial at the NCI. To investigate the role of NY-ESO-1 immunohistochemistry in patient selection and gain better insight into the incidence of NY-ESO-1 expression in synovial sarcomas and other mesenchymal tumors, we evaluated NY-ESO-1 expression by immunohistochemistry in 417 tumors. This collection of samples included: 50 SS18/SSX1/2 fusion positive synovial sarcomas, 155 gastrointestinal stromal tumors (GIST), 135 other spindle cell sarcomas as well as 77 other sarcomas (chondrosarcoma, osteosarcoma, dedifferentiated liposarcoma, alveolar soft part sarcoma, rhabdomyosarcoma, angiosarcoma, malignant mesothelioma, and Ewing’s sarcoma). We report that 76% of synovial sarcomas expressed NY-ESO-1 in a strong and diffuse pattern (2−3+, >50–70% of tumor cells). In contrast, only rare cases of other spindle cell mesenchymal tumor expressed NY-ESO-1 (GIST (2/155), malignant peripheral nerve sheath tumors (1/34), and dermatofibrosarcoma protuberans (2/20)). Individual cases of other sarcomas (angiosarcoma, malignant mesothelioma, chondrosarcoma, osteosarcoma, dedifferentiated liposarcoma, alveolar soft part sarcoma, and Ewing’s sarcoma) were positive for NY-ESO-1. However, no positive cases were identified amongst our cohort of leiomyosarcomas (0/24), hemangiopericytoma/solitary fibrous tumors (0/40), and cellular schwannomas (0/17). In summary, we find that NY-ESO-1 is strongly and diffusely expressed in a majority of synovial sarcomas, but only rarely in other mesenchymal lesions. Beyond its role in patient selection for targeted therapy, immunohistochemistry for NY-ESO-1 may be diagnostically useful for the distinction of synovial sarcoma from other spindle cell neoplasms.

Tumor associated antigen specific T-cell populations identified in ex vivo expanded TIL cultures

Ex vivo expanded tumor infiltrating lymphocytes (TILs) from malignant melanoma (MM) and head & neck squamous cell carcinoma (HNSCC) share a similar oligoclonal composition of T effector memory cells, with HLA class I restricted lysis of tumor cell lines. In this study we show that ex vivo expanded TILs from MM and HNSCC demonstrate a heterogeneous composition in frequency and magnitude of tumor associated antigen specific populations by Elispot IFNγ quantitation. TILs from MM and HNSCC shared reactivity towards NY ESO-1, cyclin B1 and Bcl-x derived peptides. Additionally we show that dominating T-cell clones and functionality persists through out expansion among an oligoclonal composition of T-cells. Our findings mirror prior results on the oligoclonal composition of TIL cultures, further indicating a potential for a broader repertoire of specific effector cells recognizing the heterogeneous tumors upon adoptive transfer; increasing the probability of tumor control by minimizing immune evasion by tumor cell escape variants.

Expression and clinical significance of NY-ESO-1 protein in human laryngeal squamous carcinoma

Objective To detect the expression of Cancer-Testis antigen NY-ESO-1 in human laryngeal squamous carcinoma (LSC) and to explore its significance in immunotherapeutic application. Methods The expressions of NY-ESO-1 protein in the LSC and in the pathologically positive lymph nodes were detected by PV-9000 Immunohistochemistry. Western blotting was also employed to measure the expressions of NY-ESO-1 in the tumor core region(TC), the tissues at the sites of 0.5cm, 1.0cm away from LSC periphery and the distant normal larynx tissues. Results NY-ESO-1 protein expression was positive in 30 out of 69 (43.48 %) cases of LSC. The expression level of NY-ESO-1 protein were found to significantly decrease by tums in TC and corresponding adjacent tissues (P ＜0.01). None of the nine normal larynx tissues expressed NY-ESO-1 protein.It did not display an obvious correlation between the expression of NY-ESO-1 with T staging, pathological grading and lymph node metastasis (P ＞0.05). Its positive expression was found in pathologically positive cervical lymph nodes, which were significantly lower than that in the primary site (P ＜0.05). Conclusion NY-ESO-1 protein express at high level in human laryngeal squamous carcinoma, and they may play a role in genesis and development of tumors, which suggests that NY-ESO-1 gene might be used as target antigens for immunotherapy of LSC and the further research is necessary. Key words: Laryngeal neoplasm; Gene expression; Lymph nodes; NY-ESO-1 protein

Abstract LB-175: Tumor regression in patients with metastatic synovial sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1

Abstract Discussant: To be announced. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-175.

The Epigenetic Therapies Azacitidine and Sodium Valproate Augment Immune Responses to the MAGE Cancer Testis Antigen in Acute Myeloid Leukemia and Myeloma.

Abstract 2086Poster Board II-63The mechanism by which epigenetic therapies exert an anti-tumor effect in Acute Myeloid Leukemia (AML) and myeloma is unknown. We have studied immune responses to cancer testis antigens (CTA) a family of immunodominant tumor-associated antigens, in 12 patients with AML treated with the DNA methyltransferase inhibitor 5-Azacitidine (AZA) and the histone deacetylase inhibitor sodium valproate (VPA). The frequency of CTA-specific CD8+ T cells was studied in 12 patients with high risk AML prior to treatment with AZA (75 mg/m2 × 7 days per 28 day cycle) and sodium valproate (1-2.5 g daily administered continuously as tolerated) on a Phase II clinical trial. CTA specific CTLs were detectable in 4 of 12 patients with relapsed AML undergoing treatment with AZA and VPA. In one patient CTLs specific to the peptide MAGE-A2157-166 were detectable prior to commencement of trial therapy at a frequency of 0.001%.and persisted throughout treatment. In three patients CTLs were only detectable after commencement of AZA therapy. A polyclonal T cell line was subsequently generated from one patient and tetramer staining of the line revealed the presence of 1% MAGE-A2157-166-specific T cells and this could be enriched using anti-PE magnetic beads (Miltenyi Biotec). Two patients in whom AZA induced a CTL response to MAGE achieved a partial response to trial therapy by Cheson criteria. Treatment with AZA resulted in up to 100fold increased levels of expression of MAGEA4, MAGEA1, HAGE, SSX1, NY ESO1, MAGEC1, LAGE and XAGE mRNA in AML cell lines: HL60, KG1a, U937 and NB4 and multiple myeloma cell lines: U266 and JJN3. We next studied whether prior treatment with AZA augmented recognition of hematopoietic tumors by MAGE specific T cell clones. Prior treatment with AZA increased recognition of two myeloma cell lines (JJN3 and U266) by a MAGEA1 specific T cell clone. This is the first demonstration that AZA induce a CTL response to MAGE in patients with AML. Taken together our data are consistent with the possibility that epigenetic manipulation of cancer testis antigen expression and induction of a tumor specific CTL response may contribute to the clinical activity of AZA in hematologic malignancies. Disclosures:No relevant conflicts of interest to declare.

CT抗原を用いた特異的免疫療法

CT抗原を用いた特異的免疫療法

Phase II trial of vaccination with full length NY-ESO-1/IMX in patients with advanced malignant melanoma

2571 Background: NY-ESO-1 is an immunogenic “cancer-testis” (CT) antigen, expressed in many cancers including melanoma. Iscomatrix adjuvant (IMX), a saponin-based adjuvant, was formulated with NY-ESO-1 (ESO) protein (ESO/IMX). ESO/IMX was highly immunogenic as defined by ESO specific antibody (Ab), delayed type hypersensitivity (DTH) &amp; T-cell responses in patients (pts) with resected ESO+ve cancer. Disease free survival in vaccine recipients with resected melanoma appeared longer than in controls. A prospective evaluation was subsequently undertaken to determine whether ESO/IMX could mediate antitumor responses in pts with advanced NY ESO-1+ve melanoma. Methods: 27 pts were vaccinated in a trial designed to assess objective clinical responses, safety, &amp; immunogenicity. Vaccination consisted of 100 μg ESO/IMX administered q 4 wk × 3; this cycle was repeated in pts without symptomatic progression unless they required other treatment. Pts were then eligible to continue vaccination q12 weeks so long as they were responding or stable. Results: ESO/IMX was well tolerated. 13 pts progressed when evaluated after the first cycle at week 11. No objective clinical responses were seen. Vaccine-induced antibody titers to NY-ESO-1 were comparable to those seen in the earlier minimal residual disease trial (ranging to &gt; 1: 390,000). DTH &amp; T-cell responses were less marked. This occurred despite many pts with advanced measurable disease having pre-existing spontaneous ESO immunity ( Table ). DTH responses were seen in 11 pts; 5 had pre-existing responses (2 were boosted) and 6 were induced. The DTH responses were lost by week 33. Conclusion: No objective responses were seen. Vaccine-induced immunity appeared to be attenuated in the presence of advanced metastatic disease. Ongoing laboratory studies are investigating the role of regulatory T cells in the suppression of ESO-specific immunity in these pts with a view to adopting strategies to counter regulatory responses e.g. low-dose cyclophosphamide prior to vaccination. [Table: see text] [Table: see text]

Expression of NY-ESO-1 and LAGE-1 cancer-testis antigens in hepatocellular carcinoma

To study the role of NY-ESO-1 and LAGE-1 cancer-testis antigens as targets for immunotherapy and the relationship between corresponding gene expression and biologic behavior of hepatocellular carcinoma (HCC). The expression of NY-ESO-1 and LAGE-1 was studied in frozen tumor tissues from 30 cases of HCC by reverse transcriptase-polymerase chain reaction and immunohistochemistry. NY-ESO-1 expression and its distribution were further studied by immunohistochemistry in a tissue array contained 191 cases of HCC. NY-ESO-1 and LAGE-1 mRNAs were expressed in 33.3% (10/30) and 16.7% (5/30) of HCC respectively. Either NY-ESO-1 or LAGE-1 was expressed in 36.7% (11/30) cases. NY-ESO-1 was expressed mainly in the cytoplasm of tumor cells. It was positive in 13.8% (24/174) cases of HCC. There was an increased expression of NY-ESO-1 from 6.8%, 3/44 in small HCC, 16.2%, 21/130 in advanced HCC and 23.1%, 12/52 in metastatic HCC. The expression in the non-metastatic group was 9.8% (12/122). The differences between the metastatic group and non-metastatic group (< 0.05) and between normal liver tissue and HCC (< 0.01) were statistically significant. There was no relationship between NY-ESO-1 expression and tumor size. NY-ESO-1 and LAGE-1 were not detected in adjacent normal liver tissue. NY-ESO-1 and LAGE-1 are expressed in a high percentage of HCC, especially in cases with metastasis. It is thus possible that NY-ESO-1/LAGE-1 can serve as targets for antigen-specific immunotherapy in HCC and NY-ESO-1 peptide vaccination may be of use for patients with advanced HCC.

Primary malignant melanoma of the oesophagus: a clinical and pathological study with emphasis on the immunophenotype of the tumours for melanocyte differentiation markers and cancer/testis antigens.

Primary malignant melanomas of the oesophageal squamous mucosa are exceedingly rare. We present here the clinical and pathological findings of 10 patients (mean age 64 years) with primary oesophageal melanoma, with emphasis on the immunophenotype of the tumours. The majority of melanomas were located in the mid to distal oesophagus and were large (mean tumour size at the time of diagnosis 6.2 cm; mean depth of invasion 1.86 cm). All but two of the melanomas were associated with an extensive in situ component. Half of the tumours were amelanotic. The histological spectrum was wide, including appearances mimicking lymphoma, poorly differentiated adenocarcinoma or sarcoma. Immunohistochemical studies were performed on six tumours using monoclonal antibodies (MAb) to S100 protein, tyrosinase (MAb T311), Melan-A (MAb A103), and gp100 (MAb HMB-45), as well as antibodies to five cancer/testis (CT) antigens (MAb CT7-33 to CT7/MAGE-C1, MAb ESO121 to NY-ESO-1, MAb 57B to MAGE-A4, MAb MA454 to MAGE-A1, and MAb M3H67 to MAGE-A3). Seven patients had metastatic disease at the time of presentation. All but one patient underwent resection of the tumour with negative surgical margins. Survival was poor, with a mean survival of 19.8 months. One patient, however, whose tumour was limited to the submucosa, is still alive 108 months post-oesophagectomy. All six melanomas examined by immunohistochemistry were positive for all the melanocyte differentiation markers tested. In addition, they were all positive for CT antigens, with MA454 being the most commonly found, suggesting that CT antigens may be a promising immunotherapeutic target for oesophageal melanomas.