Active Specific Immunotherapy of Malignant Melanoma with Anti-idiotypic Monoclonal Antibodies

In this study, we have utilized the human high molecular weight-melanoma associated antigen (HMW-MAA) as a target for active specific immunotherapy with mouse anti-idiotypic (anti-id) monoclonal antibodies (mAb) in patients with malignant melanoma. After having summarized the characteristics of HMW-MAA which account for its selection as a target for immunotherapy, we describe the development and characterization of mouse anti-id mAb MK2-23 which bears the internal image of HMW-MAA. Furthermore, we describe the results of the first clinical trial performed with mouse anti-id mAb MK2-23 in patients with malignant melanoma.     

Polysialogangliosides Expressed by Amelanotic Melanoma: A Possible Explanation for the Poor Response to Anti-monosialoganglioside Antibody 202 in a Patient with Melanoma

A 52-year-old Japanese woman developed numerous amelanotic metastatic melanomas on the skin and in various organs three years after a surgical operation for primary melanoma on the right axilla. The patient was treated with monosialoganglioside specific monoclonal antibody 202; however, no apparent clinical effects were observed. Ganglioside analysis of a metastatic tumor demonstrated that it expressed GM3, GM2, GD3, GD2, and polysialogangliosides. Since polysialogangliosides rarely appear in melanomas, their expression may explain the patient's poor response to MAb 202. The relationship between ganglioside composition and the effect of anti-ganglioside monoclonal antibody is discussed.     

白癜风患者血清中抗黑素细胞膜表面抗原自身抗体的检测

目的：检测白癜风患者血清中的抗黑素细胞抗体并分析其与疾病活动性及亚型之间的关系。方法：采用体外培养的正常人黑素细胞,使用活细胞ELISA检测了334例不同亚型及病期的白癜风患者血清中抗黑素细胞膜表面抗原IgG自身抗体。结果：活动期寻常型白癜风患者血清有67％检出抗黑素细胞IgG抗体,稳定期寻常型白癜风患者IgG抗黑素细胞抗体阳性率为30．3％;活动期节段型白癜风阳性率为40．54％,稳定期节段型白癜风的阳性率为21．42％。活动期寻常型白癜风患者血清中抗黑素细胞IgG抗体水平明显高于稳定期寻常型白癜风、活动期节段型白癜风患者及正常人;活动期节段型白癜风患者抗黑素细胞水平亦明显高于正常人。结论：白癜风患者体内广泛存在抗黑素细胞自身抗体,自身免疫机制可能在寻常型和节段型的发病中起一定作用。     

The use of electrochemotherapy in combination with immunotherapy in the treatment of metastatic melanoma: a focused review

Electrochemotherapy (ECT) is a treatment modality that combines low-dose chemotherapy with electroporation, thereby enhancing cytotoxicity. ECT was first utilized in the treatment of metastatic head and neck cancer. Today it is used as a local treatment for the cutaneous and subcutaneous metastases of a variety of cancers, including melanoma. In addition, recent evidence indicates that ECT in combination with immunotherapy can lead to a systemic tumor response. This review aims to summarize the efficacy of ECT in the treatment of metastatic melanoma, with a specific focus on the combination of ECT with immunotherapy.     

干扰素中和抗体对重组α干扰素治疗尖锐湿疣疗效的影响

目的观察干扰素中和抗体(NA)对重组α干扰素(IFN-α)治疗尖锐湿疣疗效的影响及临床意义。方法86例尖锐湿疣患者治疗前采用抗病毒生物中和法测定NA阴性者,经微波或CO2激光治疗后给予局部干扰素治疗,每次3×106IU,每周2次,疗程3~9个月(平均6个月),疗程中定期测定NA。结果86例患者治疗后NA阳性11例(12.79%),其中治疗后3,4,5,6及6个月后检出分别是1,2,3,2例及3例。NA阳性组HPV复发率(72.72%)高于NA阴性组(25.33%);临床治愈率(27.27%)低于NA阴性(74.67%),两组差异有显著性(P<0.01)。结论IFN-α治疗尖锐湿疣NA阳性率不高,但NA可以影响IFN-α的疗效。     

Detection of Antibodies to Human Melanoma Cells in Vitiligo and Alopecia Areata by Western Blot Analysis

Antibody reactivity to human melanoma cells (SK-Mel-23) was investigated in 48 patients with vitiligo, 14 with alopecia areata (AA), and 35 normal control individuals by Western blot analysis. Antibodies to SK-Mel-23 were found in 44 (92%) of the patients with vitiligo, in 7 (50%) of the patients with AA, and in 14 (40%) of the normal control individuals. Significant differences between patients with vitiligo and normal controls were found in the incidence and distribution of antibodies, but no significant differences were found between patients with AA and normal controls. The antibodies were predominantly directed to one or more antigens of approximately 110 KD, 103 KD, 88 KD, 70 KD, 56 KD, 46 DK, or 41 KD. The most common responses were to 110 KD, 88KD, and 70 KD antigens. These antibodies were present in 60%, 60%, and 73% of the patients with vitiligo; 7%, 14% and 35% of the patients with AA; and 11%, 11% and 40% of normal control individuals, respectively. There were no statistical differences in the incidence of antibodies to pigment cells between segmental and non-segmental vitiligo. These findings suggest that autoreactivity to pigment cells occurs mostly in patients with vitiligo and might be a secondary immune reaction to destroyed pigment cells.     

Treatment of Patients with Refractory Atopic Dermatitis Sensitized to House Dust Mites by Using Sublingual Allergen Immunotherapy

Even though atopic dermatitis (AD) is one of the most common chronic inflammatory skin diseases, its treatment remains a challenge in clinical practice, with most approaches limited to symptomatic, unspecific anti-inflammatory, or immunosuppressive treatments. Many studies have shown AD to have multiple causes that activate complex immunological and inflammatory pathways. However, aeroallergens, and especially the house dust mite (HDM), play a relevant role in the elicitation or exacerbation of eczematous lesions in many AD patients. Accordingly, allergen-specific immunotherapy has been used in AD patients with the aim of redirecting inappropriate immune responses. Here, we report three cases of refractory AD sensitized to HDM who were treated with sublingual immunotherapy.     

Late presentation of generalised bullous pemphigoid‐like reaction in a patient treated with pembrolizumab for metastatic melanoma

Dermatological toxicity is one of the most commonly reported immune‐related adverse events in patients receiving checkpoint inhibitor immunotherapy. We report the gradual development of a widespread bullous pemphigoid‐like reaction in a metastatic melanoma patient 8 months after commencing treatment with the programmed‐death‐1 (PD‐1) inhibitor pembrolizumab, requiring prolonged corticosteroid therapy. This case highlights the potential for insidious and late development of severe cutaneous toxicity following PD‐1 inhibitor therapy and suggests that even prolonged immunosuppression may not necessarily compromise the efficacy of PD‐1 inhibition in advanced melanoma.     

SLE患者血清抗磷脂抗体的分布及临床意义

目的:探讨抗磷脂抗体(APA)在SLE患者血清中的分布及临床意义。方法:应用酶联免疫吸附实验(ELISA)法检测64例正常人和56例SLE患者血清中6种APA。结果:SLE患者IgG型抗心磷脂抗体(aCL)、抗磷脂酰肌醇抗体(aPI)、抗磷脂酸抗体(aPA)、抗磷脂酰丝氨酸抗体(aPS)、抗磷脂酰胆碱抗体(aPC)的A值和阳性率均显著高于正常人;IgM型aCL、aPA、aPE、aPS、aPC的A值和阳性率均显著高于正常人。SLE时IgG型和IgM型APA的总阳性率分别为67.86%和69.65%。结论:正常人血清中存在低滴度的APA,SLE患者血清中APA的阳性率高于正常人。APA的检测对早期诊断SLE可能有一定意义。     

Lymphadenopathy detected by ultrasound examination as first diagnostic hint of chronic lymphocytic leukaemia in a patient with melanoma

Recent reports indicate that patients with malignant melanoma might be at higher risk for developing a non-cutaneous unrelated second malignancy. We describe the case of a 46-year-old woman who had a malignant melanoma on her right shoulder that was treated in 1998 by surgical excision combined with axillary lymph node dissection. In 1999, ultrasound examination of peripheral lymph nodes revealed one suspicious echopoor structure in the woman's right axilla that was not palpable. Diagnostic excision and histopathological examination revealed a small B-cell lymphocytic lymphoma, and further investigations led to a diagnosis of chronic lymphocytic B-cell leukaemia (B-CLL). We would like to point out the value of high-resolution ultrasound examination in the follow-up of patients with malignant melanoma; this examination can detect early metastasis as well as other unrelated malignancies.     

ELISA和明胶微粒凝集试验检测麻风病人及接触者血清抗体

用半合成酚糖脂抗原(NT-O-BSA)间接酶联免疫吸附试yan(ELISA)和半合成爵糖脂抗原(NT-P-BSA)致敏明胶微粒凝集试验(MLPA).检测贵州省12县麻风病人密切接触者血清抗体1715例,现症麻风病人137例:结果:接触者阳性率ELISA和MLPA分别为17.84%和13.94%(R=4.8514,P=0.0004);麻风病人阳性率分别为91.97和89.78% (X²=0.0083,P>0.5,在活动性肺结核病人及正常人的结果相同和相近.上述结果表明ELISA和MLIPA具有较高的敏感性和较好的特异性,且接触者血清抗体阳性率与各地麻风病患病率和发病率呈显著正相关.提示接触者抗体测定除可作为检测麻风菌感染外也许还可预测麻风病的流行.     

HPV6感染的尖锐湿疣患者血清抗体反应研究

目的　探讨我国HPV 6感染的尖锐湿疣患者血清抗体反应 ,以评价我国HPV感染的血清学诊断方法 ,并为疫苗研制提供理论依据。方法　采用重组杆状病毒 昆虫细胞系统制备HPV病毒样颗粒 ,通过ELISA法检测 3组共136份标本的血清抗体。结果　尖锐湿疣组血清抗体最高 (阳性率 75 .0 % ,OD平均值为 0 .111± 0 .0 94) ,较健康对照组(阳性率 2 .8% ,OD平均值 0 .0 12± 0 .0 2 4)和宫颈癌组 (阳性率 14 .3 % ,OD平均值 0 .0 2 9± 0 .0 2 2 )差异均有显著性 (P <0 .0 1)。结论　尖锐湿疣患者体内可检测到型特异性抗体 ,尤以女性和复发患者为显著 ,HPV 11感染与HPV 6b血清抗体相关 ,提示HPV病毒样颗粒可用作HPV感染的血清学诊断。     

Serum levels of leptin receptor in patients with malignant melanoma as a new tumor marker

Leptin is known to be abnormally expressed in a variety of cancers, and leptin receptors have been reported to be expressed on human melanoma cells. In this study, we evaluated the possibility that the serum levels of leptin receptor could be a tumor marker of malignant melanoma (MM). Serum samples were obtained from 71 patients with MM, and the serum levels of leptin receptor were measured by double‐determinant ELISA. Interestingly, serum levels of leptin receptor decreased gradually with the stages of MM, being highest at in situ and lowest at stage IV. There was also a trend of reverse correlation between tumor thickness and serum levels of leptin receptor. To our knowledge, this is the first report investigating the serum levels of leptin receptor in MM, and serum leptin receptor levels may be used as a useful tumor marker of MM.     

Melanoma risk alleles are associated with downregulation of the MTAP gene and hypermethylation of a CpG island upstream of the gene in dermal fibroblasts

Several association studies and GWAS on melanoma skin cancer risk have reported statistically significant signals on 9p21.3 region, where MTAP gene maps. None of the associated SNPs identified in these studies lie in the coding region of the gene and the causative relation of risk alleles with melanoma predisposition has not been elucidated. MTAP has a tumor suppressor activity and epigenetic silencing has been described in melanoma cell lines. In the present study, we show that melanoma risk alleles correlate with a MTAP allele‐specific hyper‐methylation and down‐regulation of gene expression.     

Cognitive training with photographs as a new concept in an education campaign for self‐detection of melanoma: a pilot study in the community

Background Cognitive education using only photographs has been shown to be more effective than the ABCD algorithm to improve melanoma recognition in the general population. Objective The aim of this study was to assess the feasibility and acceptability of a campaign based on cognitive education by photographs instead of the usual ABCD algorithm. Methods Brochures including a panel of photographs of melanoma and moles were mailed, supported by a media campaign and followed by a telephone survey of a representative sample of the district population. Results Among the 855 people interviewed, 38% recalled the campaign and 13% had actually read the brochures. Among the 13%, 99% found them useful, 52% reassuring and 40% anxiety provoking, while 92% agreed that using images of skin cancer was the best way to recognize melanoma. Sixty‐four percent declared that this campaign had prompted them to examine their own moles, 39% were prompted to check moles of family members, and only 11% would immediately visit a doctor. Conclusion This pilot study shows that a melanoma campaign based exclusively on images is well accepted in the community, and is associated with a favourable impact on self‐surveillance, while limiting overflow of unjustified consultations. We recommend this innovative concept be applied to large scale campaigns.     

Role of sentinel lymph node biopsy as a staging procedure in patients with melanoma: A critical appraisal

Worldwide, sentinel node (SN) biopsy for accurate staging is now part of the standard work‐up of patients with melanomas ≥1.0 mm Breslow thickness, as it is for staging patients with breast cancer. Nuclear medicine imaging and surgical techniques have evolved to such a degree that a SN can be identified and removed in virtually every patient. Nevertheless, some opposition to a routine SN biopsy remains, perhaps due to a failure to appreciate the serious implications of incomplete or inaccurate staging. Guided by a critical appraisal of the available evidence, this review elucidates the definition of an SN, discusses the sensitivity and specificity of the information it provides, emphasises that it is a minor staging procedure that can lead to improved survival when followed by appropriate therapy, and explains the necessarily unconventional and complex design of the only randomised trial that addresses this subject. It also describes other benefits and risks of an SN biopsy and outlines its role in current melanoma management.     

Killing efficacy of a new silicon phthalocyanine in human melanoma cells treated with photodynamic therapy by early activation of mitochondrion-mediated apoptosis

Photodynamic therapy (PDT) is a promising therapeutic modality that utilizes a combination of a photosensitizer and visible light for the destruction of diseased tissues. Using human-pigmented melanoma cells, we examined the photokilling efficacy of new silicon-phthalocyanines (SiPc) that bore bulky axial substituents. The bis(cholesteryloxy) derivate (Chol-O-SiPc) displayed the best in vitro photokilling efficacy (LD(50) = 6-8 x 10(-9) M) and was seven to nine times more potent than chloro-aluminium Pc (ClAlPc), a known photosensitizer used as a reference. Although Chol-O-SiPc was half as potent as ClAlPc for promoting photo-oxidative membrane damage in a cell-free assay, early events of mitochondrion-mediated apoptosis upon PDT were triggered much faster, as demonstrated by kinetics studies examining cells with permeabilized mitochondrial membranes, cytochrome c release and caspase-9 activation. Inhibition of caspase-9 activity by a substrate analogue argued for its central role in the proapoptotic events leading to cell death by Chol-O-SiPc PDT. In addition, immunoblots showed that Bcl-2 antiapoptotic oncoprotein was not a primary target of Chol-O-SiPc in M3Dau cells treated with PDT. Conclusively, Chol-O-SiPc is a useful new photosensitizer with the property of triggering cell apoptosis mediated by mitochondria.     

Impaired responses of peripheral blood mononuclear cells to T-cell stimulants in alopecia areata patients with a poor response to topical immunotherapy

BACKGROUND: Topical immunotherapy with a contact allergen is effective in alopecia areata (AA). However, the mechanism of the effect is still unknown, and pretreatment prediction of the outcome of therapy in each patient remains difficult. OBJECTIVES: To predict the clinical effect of this therapy in AA patients, we investigated the relationship between clinical responses to topical immunotherapy and in vitro proliferative responses of peripheral blood mononuclear cells (PBMC) to T-cell stimulants. METHODS: PBMC were taken from 67 AA patients before or during diphenylcyclopropenone immunotherapy and from 14 healthy controls, and proliferative responses to phytohaemagglutinin and staphylococcal enterotoxin B were evaluated by measuring [3H]-thymidine incorporation. RESULTS: PBMC from the AA patients with a good clinical response to immunotherapy showed a normal level of proliferation, whereas PBMC from the poor responders showed a markedly suppressed proliferative response and interleukin (IL)-2 production, but increased IL-4 production compared with the controls. CONCLUSIONS: The proliferative response of PBMC to T-cell stimulants may be one of the indicators of the clinical effect of topical immunotherapy for AA.     

Congenital nevi versus metastatic melanoma in a newborn to a mother with malignant melanoma – diagnosis supported by sex chromosome analysis and Imaging Mass Spectrometry

A 37‐year‐old pregnant woman presented with a 2‐cm irregular reddish nodule on her left upper arm during pregnancy. A biopsy from the lesion showed a 2.2‐mm thick malignant melanoma with intravascular invasion, 25 mitosis/mm² and no ulceration. Following induction of labor, the patient underwent re‐excision with sentinel lymph node biopsy. This showed no residual melanoma and no lymph node metastasis. The newborn boy had multiple pigmented lesions on the trunk, some of which were large and irregular. Two were biopsied and histologic examination showed dense dermal proliferation of medium sized melanocytes with multiple mitotic figures and no maturation with their descent into the dermis, raising suspicion of transplacental metastases. Examination of the placenta failed to show metastatic lesions. Multiplex polymerase chain reaction (PCR)‐based genotyping, including testing for amelogenin locus for sex chromosome determination, demonstrated the presence of Y chromosome material in the melanocytes of the newborn's lesions excluding maternal origin. A diagnosis of congenital nevi was rendered. Subsequently, Imaging Mass Spectrometric analysis of the mother's lesion showed proteomic signature expression indicative of malignant melanoma, whereas the two lesions in the newborn showed changes indicative of nevi. This case demonstrates the utility of genotyping and Mass Spectrometry analysis in this challenging clinical scenario