Increased regulatory T‐cell frequencies in patients with advanced melanoma correlate with a generally impaired T‐cell responsiveness and are restored after dendritic cell‐based vaccination

Please cite this paper as: Increased regulatory T‐cell frequencies in patients with advanced melanoma correlate with a generally impaired T‐cell responsiveness and are restored after dendritic cell‐based vaccination. Experimental Dermatology 2010; 19 : e213–e221. Abstract: Naturally occurring CD4⁺ CD25⁺ regulatory T‐cell (Treg) activity is assumed to facilitate tumor development and progression. To elucidate the possible role of Tregs in the course of melanoma progression, we analysed the frequency of Tregs in the peripheral blood of patients at melanoma stages I–IV and in patients at melanoma stage IV that underwent dendritic cell (DC)‐based immunotherapy. Using CD25, Foxp3, CD127 and HLA‐DR as Treg associated markers, we observed increased Treg frequencies in patients at the late melanoma stage (stage IV) when compared to healthy donors. Accumulation of Tregs in patients with progressed melanoma correlated with a general reduction of T‐cell responsiveness to the recall antigens tetanus toxoid and tuberculin‐GT. However, DC‐based immunotherapy not only restored antigen‐specific immunity, but also decreased the frequency of Tregs in peripheral blood of patients with melanoma. These findings indicate that tumor progression in patients with melanoma result in general immunosuppression that is associated with Treg expansion in the periphery and can be overcome by DC‐based vaccination.     

Statistical Survey from 1982 to 1991 of 49 Patients with Malignant Melanocytic Tumors

We investigated the clinical characteristics and outcome of 49 patients with malignant melanoma: 17 with acral lentiginous melanoma, 14 with nodular melanoma, six with superficial spreading melanoma, one with lentigo maligna melanoma, eight with melanoma in situ, one with malignant blue nevus, and two with melanoma of unknown origin. Of the 41 patients without melanoma in situ, 34.1% were in stage I, 17.1% in stage II, and 48.8% in stage III. No patients had reached stage IV. All patients with stage I, II, and III melanoma were treated with wide resection, lymph node dissection including prophylactic dissection, and combination chemotherapy with dacarbazine, nimustine hydrochloride, and vincristine (DAV) with or without Interferon-β. The statistical analysis revealed that tumor thickness and level of invasion were factors significantly associated with outcome. A gradual increase in the number of new cases of melanoma was seen each year of the registration period (1982–1991); there was an approximately 4-fold increase during this decade over the previous decade. The majority of the primary lesions (80.5%) were located on the upper or lower extremities, the pT4 tumor thickness subgroup was the most frequent (39.0% of all melanomas), and invasion level IV was the most common (42.5%). About half (51.2%) of the melanoma patients were stage I or II; this group had a 5-year survival rate of 100%. The stage III patients had a 5-year survival rate of only 54.2% (p<0.05).     

Comparison between familial and sporadic cutaneous melanoma in Valencia, Spain

Some clinical, pathological and genetic features have been associated to familial melanoma, particularly multiple melanoma and earlier age at diagnosis.
To compare the clinical, epidemiological and pathological differences between familial and sporadic melanoma patients in Valencia, Spain, a series of 959 patients with cutaneous melanoma were selected at a single institution. For this study the following variables were selected: age, sex, melanoma site and presence of solar lentigines on the melanoma surrounding skin, histological subtype, tumor thickness, stage, family and personal history of cutaneous melanoma and of other neoplasias, personal history of non-melanoma skin cancer, past personal history of severe sunburns, cutaneous phenotype (phototype, hair and eyes colors number of common nevus, number of atypical nevi, presence of solar lentigines).
Forty-one (4.28%) familial and 918 sporadic melanoma were identified. Among the multiple variables studied, a younger age at diagnosis (median age of 42 vs 53 years), higher frequency of the presence of at least one clinically atypical nevus (36.1% vs 17.7%), multiple melanomas (12.2% vs 3.4%) and red/blonde hair (33.3% vs 18.9%), and a lower rate of cases with solar lentigines in melanoma site (33.3% vs 56.3%) were found for familial cases. Except for hair color and age, the other variables remained statistically significant after the multivariate study. Interestingly, no acral melanomas were found among the familial cases.
In summary, phenotypic risk factors for familial melanoma are a tendency to develop multiple melanomas, to have clinically atypical nevi and to present less actinic damage at the melanoma site. All these results enhance the relevancy of genetic susceptibility associated to the ability to produce atypical nevi and partly to a higher sensitivity to the sun.     

Xenovaccinotherapy for melanoma

The objectives of this phase I-II trial were to assess the toxicity, immunological and clinical responses induced in stage III/IV melanoma patients by the subcutaneous administration of xenogenic polyantigenic vaccine (XPV) prepared from disrupted murine melanoma (B16) and carcinoma (LLC) cells. An inducing course of vaccinotherapy consisted of ten immunizations (five at weekly and five at fortnight intervals). Twenty-four hours following each of the first five vaccinations, the patient was subcutaneously given a low dose of the recombinant interleukin-2 (IL-2). A consolidating course of the vaccinotherapy consisted of monthly vaccinations. Grade 3 or 4 toxicities, as well as laboratory and clinical signs of developing autoimmune disorders, were recorded in none of the 40 XPV-treated evaluable patients. A significant increase in delayed-type hypersensitivity (DTH) skin reaction to vaccinal B16, but not to LLC antigens (Ags), occurred in patients after inducing vaccinations. At the same time, those patients demonstrated a marked augmentation of blood lymphocyte proliferation responses not only to B16 but also to LLC Ags. Vaccinations also led to increased cell-mediated reactivity to murine non-tumor, spleen cell (SC)-associated Ags, which, however, was not as significant as that to tumor-associated antigens (TAAs). Of great importance was the fact that XPV administration resulted in increased blood lymphocyte proliferative reactivity of patients to human melanoma-associated Ags, while not affecting their reactivity to the control alloantigens. With immunotherapy, concentrations of both interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) were elevated in patients' sera, suggesting an intensification of the T helper1/ T helper 2-mediated responses in the XPV-treated patients. The average survival of the 32 stage IV melanoma XPV-treated evaluable patients was noticeably higher than that of the 32 clinically comparable control patients (13 vs. 5 months). The overall 3 year-survival rate in the XPV-treated group and the control group was 25% (8 patients) and 3% (1 patient), respectively. In general, the results suggest that xenogenic tumor cells may provide a novel feasible approach to constructing clinically effective vaccines.     

Desmoplastic melanoma may mimic a cutaneous peripheral nerve sheath tumor: Report of 3 challenging cases

Desmoplastic melanoma (DM) and cutaneous malignant peripheral nerve sheath tumors (MPNST) reveal histological and immunohistochemical similarities, including S100 positivity and negative staining for conventional melanocytic markers. We present 3 cases of cutaneous S100‐positive spindle cell tumors in elderly patients, in which first findings led to initial misdiagnoses as cutaneous MPNST and benign peripheral sheath nerve tumor (neurofibroma). The identification of adjacent atypical melanocytic hyperplasia in the overlying skin along with tumor cell proliferation, also in the superficial dermis, the neurotropic component and the absence of any relationship between the tumor and a major nerve, pre‐existing neural benign tumor or the existence of stigmata suggestive of neurofibromatosis raised consideration of a DM. Careful attention should be paid to the presence of a firm dermal nodule and atypical scar lesions especially in sun‐exposed areas (mainly head and neck region) in elderly patients associated with S100‐positive spindle cell proliferation, solar elastosis and adjacent atypical melanocytic proliferation. In such cases, the possibility of a DM should be excluded with caution, especially if the tumor reveals a paucicellular morphology resembling various non‐melanocytic neoplasms including malignant or benign peripheral sheath nerve tumors.     

Familial Atypical Mole Syndrome Manifesting as Polypoid Melanoma: A Case Report and Review of the Japanese Literature

A family with atypical mole syndrome which manifested as polypoid melanoma in one member is reported. A 46-year-old man presented with a polypoid tumor on his lower back, which was excised under clinical diagnosis of soft fibroma. The tumor proved to be polypoid melanoma. He was referred to our clinic, and examination revealed that he had the atypical mole syndrome phenotype. The clinicopathological screening of his first-degree relatives confirmed that his 58-year-old sister had the same phenotype. In addition to our discussion of the clinical significance of polypoid melanoma, we also attempted to elucidate the characteristics of the reported Japanese cases of melanomas with atypical moles from a survey of the literature, which revealed the following characteristics: 1) younger mean age than that of total Japanese melanoma cases, 2) sun sensitive skin, 3) unexpectedly and frequently observed familial incidence (Kraemer's D₁ and D₂), 4) frequent site of the melanoma on the trunk and none on the sole of the foot, and 5) high incidence of superficial spreading melanoma and nodular melanoma, and 6) no cases of acral lentiginous melanoma.     

Coexistence of Amelanotic Melanoma and Liposarcoma

An amelanotic malignant melanoma is characterized by little or no pigment. It is frequently misdiagnosed because it is a rare entity in general, and because of its unusual clinical features. Liposarcoma is one of the most common adult soft tissue sarcomas. We encountered a case of amelanotic melanoma with a concurrent liposarcoma. A 68-year-old man presented with a single, 1.5×1.5 cm round erythematous, eroded nodule on the left heel. A biopsy specimen showed atypical, pleomorphic tumor cells with little melanin pigment. The tumor cells were positive for S-100, HMB-45 and negative for cytokeratins. These findings were consistent with amelanotic melanoma. On positron emission tomography/computed tomography (PET/CT), a hypermetabolic lesion was found in the left buttock. This lesion was excised and diagnosed as a well-differentiated liposarcoma. An association between sarcomas and other primary malignancies has been reported. However, an association between melanoma and liposarcoma is rare.     

Photodynamic therapy with chlorin e(6) for skin metastases of melanoma

BACKGROUND: Photodynamic therapy (PDT) has been successfully applied in clinical settings to destroy neoplasms, but the efficacy of such a treatment is dependent on the type of neoplasm and the photosynthesizer used. Here, we perform a clinical assessment of PDT for skin metastases of pigmented melanoma using chlorin e(6). STUDY DESIGN/MATERIALS AND METHODS: PDT with chlorin e(6) photosensitizer was administered to 14 patients with skin metastases from melanoma (10 females, four males, mean age 49.6 years). Chlorin e(6) at a dose of 5 mg/kg of patient's weight was intravenously injected. The treatment course consisted of two courses of PDT exposure 1 h after intravenous chlorin e(6) injection and 24 h post-injection. The light energy density for each skin tumor was 80-120 J/cm(2) per treatment, with a light power density of 250-300 mW/cm(2). RESULTS: All skin melanoma metastases that received PDT showed complete regression with no recurrence during the study period. The complete response of all skin metastases from melanoma occurred in eight cases after one PDT treatment. In the remaining six individuals, tumors required multiple PDT courses prior to complete regression. No cases of photodermatitis were registered. The Karnofsky performance scale score of the patients with skin metastases from melanoma showed no significant difference before and after PDT. No patients had significant changes in blood cell counts that would indicate chlorin e(6) systemic toxic effect. Blood chemistry and urinalysis did not show any evidence of chlorin e(6) renal and hepatic injury. CONCLUSIONS: PDT with chlorin e(6) for skin metastases from melanoma is effective and well tolerated. Further clinical investigation of PDT with chlorin e(6) is warranted.     

The current status and future direction of percutaneous peptide immunization against melanoma

Dendritic cell (DC)-based tumor immunotherapy is widely known to elicit protective anti-tumor immune responses, although the safety and effectiveness have yet to be thoroughly explored. We reported that a disruption in the stratum corneum barrier resulted in enhanced permeability and alterations in the skin immune system in such a way that epidermal Langerhans cells (LCs) functioned as vigorous antigen presenters for T helper (Th) cells and cytotoxic T lymphocytes (CTLs). In both human and murine models, topical application of melanoma-associated antigen peptides onto stratum corneum barrier-disrupted skin, specifically induced tumoricidal immune responses in vivo and in vitro accompanying an increased expression of MHC and co-stimulatory molecules on LCs. In addition, for reasons of simplicity, safety and effectiveness, percutaneous peptide application has demonstrated a certain degree of feasibility in clinical approach in patients with melanoma. In the future, resolution of some of the outstanding issues concerning the selection of the most effective adjuvants in combination with barrier disruption and depletion of regulatory T (Treg) cell-mediated immune suppression would appear as essential to improve percutaneous melanoma immunotherapy.     

Malignant melanoma treatment using brachytherapy: Two case reports and 15 case series

Malignant melanoma (MM) is usually resistant to radiotherapy. Brachytherapy may be an option in patients with bleeding or pain, and those in whom surgery is difficult. Brachytherapy has few side effects and can be used in combination with external beam radiotherapy or chemotherapy. We summarize the demographic and clinical characteristics of 15 patients who received brachytherapy for MM at our hospital and describe two of these representative cases. Patient 1 had an approximately 10-mm, dark-red nodule near the external urethral meatus. Excision was not performed to preserve urethral function. A gradual improvement was observed after 48 Gy of remote afterloading system (RALS) brachytherapy and nivolumab therapy. Patient 2 had a 38-mm, black tumor on the vagina. Post-resection, RALS brachytherapy was administered to treat the residual black macule and a lesion quickly disappeared. In all 15 cases, nine patients received radiotherapy for local control and six patients received palliative radiotherapy to reduce symptoms such as bleeding and pain. The irradiation site was the vagina in six patients, lymph node metastasis in five, head and neck in two, skin or subcutaneous metastases in two, and the anus in one. Treatment effect for local control and palliative care was 75% and 83% of patients, respectively. In particular, disappearance of the tumor or disappearance of symptoms was observed in half of the cases of brachytherapy to the vagina. On the other hand, brachytherapy was not very effective for lymph node metastases. Immediately after radiotherapy, eight (53%) patients experienced dermatitis or mucositis. Due to the histological and structural characteristics of mucosal melanoma of the luminal organs, brachytherapy may be an effective therapy. Hence, widespread use of brachytherapy with an appropriate irradiation technique aiming for local control and palliative care in case of unresectable MM should be considered.     

A rare case of primary dermal clear cell sarcoma with focal epidermotropism: An entity difficult to distinguish from melanoma

Clear cell sarcoma (CCS) is an uncommon soft-tissue sarcoma that only rarely arises within the dermis. It is challenging to distinguish dermal CCS from nodular, primary dermal, or metastatic melanoma, as they share morphologic features and immunoprofiles. We describe a dermal CCS in a 25-year-old man with a cutaneous groin mass. The lesion was initially diagnosed as melanoma, likely metastatic. On consultation, in addition to a melanoma-like tumor in the dermis, we identified focal infiltration of tumor cells into the overlying epidermis (epidermotropism), resembling primary nodular or metastatic melanoma. Given the patient's age and absence of a history of primary melanoma, fluorescence in situ hybridization (FISH) was performed, which revealed separation of the 5′ and 3′ EWSR1 probe signals on chromosome 22q12, prompting a diagnosis of CCS. Our case highlights the histopathological, immunohistochemical, and ultrastructural similarities between CCS and melanoma, and the consequent potential for major diagnostic confusion. In such cases, FISH analysis remains the key to diagnosis. CCS should be considered in patients with a melanoma-like tumor in the dermis or subcutaneous tissue without epidermal (or with minimal) involvement, or prior to diagnosing metastatic melanoma in the absence of a known history of primary melanoma, especially in young individuals.     

新疆地区58例恶性黑色素瘤临床及病理初步分析

目的探讨新疆地区恶性黑色素瘤的临床及病理特征。方法回顾性分析我院(2005～2011)年经病理及免疫组化确诊的58例恶性黑色素瘤患者病例。结果 58例恶性黑色素瘤患者中男性27例(46.6%),女性31例(53.4%),男女之比:1∶1.15;发病年龄(22～94)岁,平均57.5岁,其中≤40岁8例(13.8%),(40～60)岁23例(39.7%),≥60岁27例(46.5%);发病部位在皮肤42例(72.4%),黏膜16例(27.6%),其中25例(43.1%)原发部位在四肢,其中上肢7例,下肢18例;58例患者44例(75.9%)行手术治疗,其中19例(43.2%)术后联合放化疗;病理形态上肿瘤以多种细胞类型和组织形态混合存在为主要特点,且58例患者均经免疫组化确诊。结论该地区恶性黑色素瘤多为中老年人,性别差异小,四肢皮肤为好发部位,其中下肢发病高于上肢;恶黑临床表现多样化,组织结构复杂,确诊主要依赖病理检查确诊,目前治疗仍以手术为主。     

Dendritic Cell‐Based Immunotherapy of Malignant Melanoma: Success and Limitations

Dendritic cells (DC) are professional antigen‐presenting cells in the immune system which are able to induce primary T‐cell responses. Because of their central role in the initiation of immune responses, DC are an important tool for tumor‐antigen‐specific immunotherapy of cancer. DC vaccination using tumor‐antigen‐loaded DC has led to tumor regression in individual advanced‐stage cancer patients. However, there is a discrepancy between strong and antigen‐specific T cell responses in vaccinated cancer patients detectable ex vivo and only weak clinical responses. In most cases the immune system of advanced stage IV cancer patients allows only a temporary anti‐tumor response and increasing evidence exists that active suppressive mechanisms of the immune system as well as of the tumor itself ultimately prevent “autoaggressive“ immune reactions against the tumor. Active counter‐regulation of effector T cells by tumor‐antigen‐specific regulatory T‐cell (Treg) populations play a central role in limiting the efficacy of the vaccines. Nevertheless, recent studies have shown that DC,additionally activated byToll‐Like‐receptor ligands (TLRL) can neutralize these suppressive effects of Treg and facilitate the induction of long‐lasting effector T cell responses even in the presence of activated Treg. These studies open a new way for “conditioning“ of DC by TLRL and might significantly enhance the efficiency of DC‐based melanoma vaccines in the future.     

Nodular malignant melanoma with Spitz nevus-like pathological features finally confirmed by the pathological feature of the sentinel lymph node

The clinical and histopathological similarities of nodular melanoma and Spitz nevus currently still make a definitive diagnosis difficult. We report here a case of nodular melanoma that was extremely difficult to diagnose both clinically and histopathologically. The primary tumor was a blackish nodule on the scalp and biopsy was performed for pathological diagnosis. Although our first impression was malignant melanoma, we asked two dermatopathologists for second opinions; however, one diagnosed a melanoma and the other a Spitz nevus. Faced with this clinical dilemma, an operation was performed with sentinel node biopsy. Only one sentinel node suggested a metastasis. Histopathological diagnosis to establish whether it was a melanoma metastasis or nodal nevi was also difficult, and we again asked for second opinions from another dermatopathologist in the USA. According to its clinical course and the histopathology of the sentinel lymph node with additional immunohistochemistry, this case was finally diagnosed as a nodular melanoma (T4aN1aM0, stage IIIA). To date, the patient has been given five courses of chemotherapy at 6-month intervals, with no local recurrence or distant metastases so far.     

Pagetoid malignant melanoma and lentigo maligna melanoma of toe

Summary Two cases of malignant melanoma on the toe of middle-aged women were examined chiefly by the fluorescence method of Falck and Hillarp. In one of the patients, histopathology of the pigmented tumor on the left middle toe was a Pagetoid (superficial spreading) melanoma in situ, and the subungual granulomatous lesion on the right great toe in the other patient was a lentigo maligna melanoma. On fluorescence microscopy, characteristic findings of the pigment cells lying in the epidermis of both types may be summarized as follows: In the Pagetoid melanoma, the melanoma cells are ovoid, lack dendritic processes, and emit specific yellow fluorescence. In the lentigo maligna melanoma, the pigment cells clearly show dendritic processes, and emit specific green fluorescence.     

A novel surgical margin (1 cm) might be from benefit for patients with dysplastic nevi,thin melanomas,and melanoma in situ: Analysis based on clinical cases

Malignant melanoma is an oncologic disease, whose current management among others includes surgical and immunological therapy. According to the current recommendations of the American Joint Committee on Cancer, the surgical excision of the primary tumor should be performed in two operative sessions, which has several consequences. The following paper will present and discuss six cases of pigmented lesions and the advantages of the one‐step melanoma surgery in their management.     

PD‐L1 expression by tumor cell lines: A predictive marker in melanoma

Prognostic biomarkers for patients with melanoma after lymph node resection are of clinical relevance and could thus enable the identification of patients who therefore would most benefit from adjuvant treatment. The aim of this work was to determine, using an in vitro model, whether immune‐related biomarkers, such as MHC‐class I and II, melanoma‐associated antigens, IDO1 and PD‐L1, could also be relevant to predict the risk of relapse of patients with stage III melanoma after lymph node resection. We established tumor cell lines from metastatic lymph nodes of 50 patients with melanoma. The expression of investigated biomarkers was determined on untreated and IFN‐γ treated melanoma cell lines using flow cytometry. Among the selected biomarkers, the IFN‐γ‐induced expression of PD‐L1 and IDO1 was associated with an increased risk of relapse (P = .0001 and P = .013, respectively) and was also associated with death for IDO1 (P = .0005). In the future, this immunologic signature could permit the identification of patients at higher risk of relapse and justifying an adjuvant treatment using immunotherapy.