Immunohistochemical study of malignant melanoma

Fifty-two malignant melanoma cases were divided morphologically into round, spindle and mixed-cell types and were studied immunohistochemically on the localization and staining intensity of S-100 protein and neuron specific enolase (NSE). Most of malignant melanomas were positively stained for S-100 protein and NSE. There were no correlation between the localization of S-100 protein and three cell types. However S-100 protein and degree of melanin production seemed to have an inverse relationship. On the other hand, for NSE, there were some differences on immunostaining intensity among the three cell types. Especially, deeply invasive melanomas showed strong reactivities for NSE and it may clinicopathologically indicate their poorer prognosis.     

Immunohistochemical study of S-100 protein and neuron specific enolase (NSE) in melanocytes and the related tumors

Normal human skin, malignant melanoma, nevocellular nevus, blue nevus, nevus of Ota and mongolian spot were immunohistochemically investigated on the localization of S-100 protein and neuron specific enolase (NSE). Tissues were fixed with buffered-formalin, processed with routine procedure and examined by the ABC technique. All cases of malignant melanoma and nevocellular nevus showed a relatively high amount of S-100 protein, but NSE was scantly demonstrated on about the half cases of these tumors. Blue nevus, nevus of Ota and mongolian spot revealed the presence of a small amount of S-100 protein and NSE on the half cases. Normal melanocytes were devoid of S-100 protein and NSE. Our results suggest that S-100 protein is the useful marker for diagnosis of malignant melanoma, and immunoreactive intensity for S-100 protein represents the differentiation of neural crest derived melanogenic cells and tumors.     

Measuring S100 protein and neurone specific enolase in melanocytic tumours using video image analysis.

Using a computed video image analysis system, the staining intensity for both neurone specific enolase (NSE) and S100 protein was measured in sections from 19 malignant melanomas and 16 benign melanocytic lesions. The results of this study confirm previous reports that NSE and S100 protein are useful markers for malignant melanoma. NSE staining intensity in the cases of malignant melanoma was significantly higher than that in benign naevi (p = 0.011). Intensity of staining for S100 protein was not significantly higher in the malignant melanomas. There was, however, a significant S100 gradient when comparing superficial and deep intradermal portions of these tumours (p = 0.003). This feature was not seen in benign naevi. The greatest intensity of S100 protein staining was found in the deeper portions of the malignant melanomas. This gradient difference was not seen with staining for NSE. Although it seems that the overall intensity of staining for NSE is more effective in differentiating between benign and malignant lesions, the difference in staining intensity between the superficial and deep portions of the tumour may be the better indicator of adverse behaviour in lesions in which the diagnosis of malignancy is uncertain.     

Balloon cell melanoma in three dogs: a histopathological, immunohistochemical and ultrastructural study.

Balloon cell melanoma, a variant of malignant melanoma, has been reported on rare occasions in animals and is uncommon in man. Such tumours have variable numbers of large, round to polygonal cells with abundant, clear, often vacuolated cytoplasm containing fine melanin granules and variable amounts of lipid. This report describes balloon cell melanomas in three dogs. Immunohistochemically, these tumours showed reactions similar to those of human melanomas when tested with antibodies against S-100 protein, neuron-specific enolase (NSE) and vimentin. Electron microscopically, numerous heterogeneous melanosomes were demonstrated in the balloon cell cytoplasm of one tumour. Although balloon cell melanoma apparently occurs infrequently in dogs, it should always be considered in the differential diagnosis of neoplasms containing clear cells.     

Immunohistochemical study of melanocytic differentiation antigens in cutaneous malignant melanoma. A comparison of six commercial antibodies and one non-commercial antibody in nodular melanoma, superficially spreading melanoma and lentigo maligna melanoma

In certain primary and metastatic malignant melanomas diagnostic problems may arise due to their cytologic features and/or absence of synthesis of melanin. As the "classic" combination of S-100 protein and HMB-45 may occasionally fail to stain cells of malignant melanoma, we have tested a series of commercially accessible antibodies which were so far not compared by other authors in the three most frequent subtypes of this tumor. In surgical specimens from 104 cutaneous malignant melanomas (40 nodular melanomas, 46 superficially spreading malignant melanomas and 18 lentigo maligna melanomas) the staining intensity and the proportion of neoplastic cells stained with antibodies to S-100 protein, HMB-45, NKI/C3, NKI/beteb, MART 1 (Melan A), KBA 62 and Mitf was semiquantitatively analysed. The use of this group of antibodies against melanoma-associated antigens revealed it to be a favourable supplement for the bioptical or cytological diagnosis of malignant melanoma in case the traditional/conventional combination of S-100 protein and HMB-45 antibody fails. According to the authors' experience the antibody against KBA 62 has shown to be the most effective antibody followed by the antibodies against MART-1 (Melan A) and NKI/C3.     

Typical, dysplastic, congenital, and Spitz nevi: a comparative immunohistochemical study

Nevus cell components have been observed in up to 40% of melanomas, but little is known of the pathobiology of these components in relation to their malignant potential. We studied 44 nevi of the typical, dysplastic, congenital, and Spitz types with a battery of monoclonal and polyclonal antibodies that react on formalin-fixed, paraffin-embedded tissues (HMB.45, S-100 protein, RAP-5, epithelial membrane antigen [EMA], and neuron-specific enolase [NSE]) by avidin-biotin immunohistochemical methods. EMA and RAP-5 (which detects the ras oncogene-associated P21 protein) were negative in all cases. Melanoma-specific HMB.45 was strongly reactive with the epidermal component and had a weak to negative reaction with the dermal component in the typical nevi. However, the reaction seen with HMB.45 in the junctional component of dysplastic nevi, congenital nevi, and some Spitz nevi was heterogeneous. One Spitz nevi showed HMB.45 staining in a pattern near to that of melanoma. In contrast to HMB.45, S-100 protein labeled nevomelanocytes, regardless of whether they were within the epidermis or dermis, although, in half of the dysplastic nevi, the reaction was heterogeneous, with some atypical cells failing to stain. But, with cytologically atypical junctional component (dysplastic-appearing), congenital nevi also stained heterogeneously for S-100 protein compared with the dermal component. NSE stained the central component of some Spitz nevi more intensely than the lateral component. Junctional nevomelanocytic subsets of some congenital nevi revealed HMB.45 and S-100 reactivity similar to dysplastic nevi.     

Esthesioneuroblastoma. Intermediate filaments, neuroendocrine, and tissue-specific antigens

Esthesioneuroblastoma (EN), a malignant neuroblastic tumor arising in the superior portion of the nasal cavity, shares histologic similarities with a number of primary malignant tumors that arise in this region, including rhabdomyosarcoma, lymphoepithelioma, and lymphoma. To establish an antigenic profile of EN as an aid in the differential diagnosis of these histologically similar nasal tumors, immunostaining was performed for the following intermediate filaments: keratin, neurofilament, glial fibrillary acidic protein, and desmin; neuron-specific enolase (NSE), S-100 protein, chromogranin, human common leukocyte antigen (HLE), epithelial membrane antigen (EMA), myoglobin, and carcinoembryonic antigen (CEA) on 21 primary nasal tumors: eight EN, five lymphoepitheliomas, two small cell carcinomas, three lymphomas, and three rhabdomyosarcomas. Keratin and CEA stained only the carcinomas (6/7+, 4/7+), respectively; desmin and myoglobin only rhabdomyosarcoma (3/3+, 1/3+); and HLE only lymphomas (3/3+). Chromogranin and neurofilament staining occurred exclusively in one case each of EN. S-100 and NSE commonly stained EN (5/8+, 6/8+), but carcinomas (1/7+, 2/7+) and rhabdomyosarcomas (1/3+, 3/3+) were also positive. Despite the apparent nonspecificity of NSE and S-100, an antigenic profile of positive NSE of S-100 staining with negative epithelial, muscle, and lymphoid antigens uniquely identified six of eight EN. Chromogranin and neurofilament positivity was further evidence for EN in two cases. This antigenic profile is a helpful adjunct in the diagnosis of EN and other primary malignant nasal tumors.     

Paragangliomas: assessment of prognosis by histologic, immunohistochemical, and ultrastructural techniques

To predict clinical outcome, we studied 42 paragangliomas from 37 patients by routine histology, immunohistochemistry, and electron microscopy. A panel of antisera to neuron-specific enolase (NSE), chromogranin, and met-enkephalin was used to identify chief (type I) cells, and S-100 protein and glial fibrillary acid protein (GFAP) sustentacular (type II) cells. The intensity of staining of type I cells and the density of type II cells were assessed semiquantitatively (0 to 4+) in a total of 38 tumors. A total of 23 of 24 low-grade tumors (solitary, multiple, or associated with other neoplasms; 95.8%) contained type II cells immunoreactive with either S-100 protein or GFAP, and all were positive when S-100 protein and GFAP were used in combination. Five of the nine intermediate-grade (recurrent and/or locally aggressive) tumors were identified as glomus jugulare tumors (GJT). Three intermediate-grade GJTs were devoid of GFAP-reactive type II cells and four GJTs were negative for S-100 protein. Type II cells were identified in only one of five high-grade (malignant) paragangliomas and that tumor contained vanishingly rare cells that were weakly S-100 protein positive but GFAP negative. Sustentacular cell density and chief cell staining intensity were both inversely related to tumor grade. The most sensitive chief cell marker was NSE (92.1%), followed by chromogranin (84.2%). The least sensitive (73.0%) and specific marker was met-enkephalin. Combinations of NSE or chromogranin with met-enkephalin identified chief cells in all cases. Electron microscopy identified neurosecretory granule-containing chief cells, but was of less value in delineating sustentacular cells because of their scarcity and the absence of specific features. By comparison, immunohistochemistry was superior in identifying sustentacular cells. The use of an immunohistochemical panel, in addition to routine histology, can confirm the diagnosis of a paraganglioma and can give an indication of the likely prognosis for a patient.     

Neurone specific enolase and S100 protein as possible prognostic indicators in melanoma*

Fourteen cases of primary melanoma and 25 of their subsequent metastases were stained for Neurone Specific Enolase (NSE) and S100 protein. Intensity of staining for NSE and S100 protein broadly corresponded in 11 of the primary lesions and was disparate in three. Staining intensity for NSE or S100 was independent of tumour thickness. Primary lesions showing marked or moderate staining for NSE and S100 protein took a shorter time to metastasize than those showing slight or no staining. Assessment of staining intensity for NSE and S100 thus identified prognostic categories corresponding to disease free interval obtained by division according to tumour thickness. Staining intensity for S100 protein appears to give a clearer indication as to expectation of disease free interval. Staining intensity in individual cases showed an increase both for NSE and S100 protein between primary and metastatic lesions. The data presented are not sufficient to assign statistical significance but may lead to the incorporation of functional studies into the pathological assessment of malignant melanocytic lesions. The simultaneous occurrence of a functional neuronal and Schwann cell marker in melanoma is discussed.     

Myoepitheliomas and myoepithelial adenomas of salivary gland origin. Immunohistochemical evaluation of filament proteins, S-100 alpha and beta, glial fibrillary acidic proteins, neuron-specific enolase, and lactoferrin

Immunohistochemical identification of keratin proteins (TK, KL1 and PKK1), vimentin, myosin, S-100 protein (using polyclonal antiserum) and S-100 alpha and beta subunits, glial fibrillary acidic protein (GFAP), neuron-specific enolase (NSE), lactoferrin, and lysozyme was made in myoepitheliomas, myoepithelial adenomas, and clear cell adenomas of salivary gland origin. Myoepithelioma cells were divided into two types: plasmacytoid cells, which showed great heterogeneity in terms of keratins and S-100 alpha and beta proteins and a lack of GFAP, NSE, lactoferrin, and lysozyme in most the cells, and fibrous and dendritic tumor cells, which displayed variable staining for keratin and S-100 alpha and beta proteins. Myoepithelial adenomas were composed of small-, intermediate-, and large-sized spindle cells that showed irregular positive reactions for keratins and S-100 alpha and beta. Immunohistochemical deposition of S-100 protein was restricted strongly to the dendritic cells present in hyalinous and myxomatous areas. Clear cell adenomas revealed uniformly slight staining of keratins and S-100 proteins, and negative staining or rarely positivity for GFAP, NSE, lactoferrin, and lysozyme. When the immunohistochemical deposition of these proteins was compared between normal glands and myoepithelial tumors, heterogeneity of expression of keratins, S-100 proteins, GFAP, and NSE was notable in the tumors. Progenitor cells of several kinds of myoepithelioma were suggested to be intercalated reserve cells, which are thought to be the same cell that gives rise to pleomorphic adenoma of salivary glands.     

Immunocytochemical localizations of neuron-specific proteins in the taste bud of the guinea pig

The taste buds and their nerves in the guinea pig were immunocytochemically investigated with regard to the localization of spot 35 protein, neuron-specific enolase (NSE), neurofilament protein (NFP), and S-100 protein. The spot 35 protein-like immunoreactivity was confined to slender cells comprising half the number of taste bud cells. NSE-like immunoreactivity was recognized in some taste bud cells as well as nerve fibers both within the taste bud and in the subepithelial connective tissue. The NSE-immunoreactive cells were divided into two cell populations: one weakly and the other intensely immunoreactive. The former cells proved to be spot 35 protein-immunoreactive. Moreover, the cells immunoreactive for both spot 35 protein and NSE were frequently associated with nerve fibers immunostained intensely for NSE. The NFP- and S-100 protein-like immunoreactivities were found in none of cells in the taste bud, but exclusively in the subepithelial neural elements.     

S-100 protein in soft-tissue tumors derived from Schwann cells and melanocytes.

In soft tissues outside the central nervous system, S-100 protein is found normally only in Schwann cells. Using the peroxidase-antiperoxidase immunohistochemical method S-100 was also found in tumors derived from Schwann cells and melanocytes, including neurofibromas, neurilemomas, granular cell myoblastomas, cutaneous nevi, and malignant melanomas. S-100 was not detected in malignant Schwannomas, neuroblastomas, oat cell carcinomas, medullary carcinomas of the thyroid, paragangliomas, or meningiomas. S-100 was also absent from neoplasms of soft tissues not usually considered to arise from cells of neural crest origin. S-100 appears to be a useful marker for identifying neoplasms derived from Schwann cells and melanocytes.     

Neurospecific proteins: potentials and prospects for their use in morphological research on tumors

Data on the application of neurospecific proteins S-100, GFAP, D2 glycoprotein and neuron-specific enolase (NSE) in the differential tumor diagnosis are reviewed. S-100 protein and GFAP are found in well differentiated astroglial tumors. S-100 protein can be used as melanoma and Schwannoma specific marker. In malignant CNS tumors there is a decrease of S-100 protein content up to its complete disappearance, while the content of GFAP is variable. D2 glycoprotein is detected in gliomas and medulloblastomas, being absent in other brain tumors. NSE is invariably present in apudomas and was also found in the majority of investigated astrocytomas, ependymomas, glioblastomas and in some medulloblastomas.     

Primary mucosal malignant melanoma: an immunohistochemical study of 12 cases with comparison to cutaneous and metastatic melanomas

Immunohistochemical analysis of 40 formalin-fixed, paraffin-embedded malignant melanomas (12 primary mucosal, 16 primary cutaneous, and 12 metastatic cutaneous) was performed to study the possible differences in immunostaining profiles according to location. The majority of melanomas were reactive with a polyclonal antibody to S100 protein (P-S100; 85%), a monoclonal melanoma-specific antibody (HMB-45; 88%), and a monoclonal antibody to vimentin (90%), and there were no differences in staining profiles for these antibodies by anatomic location. In contrast, while 13 of 16 cutaneous melanomas (81%) and ten of 12 metastatic melanomas (83%) were reactive with a monoclonal antibody to S100 protein (MoAb-079), only five of 12 mucosal tumors (42%) showed positive staining for MoAb-079. Similarly, 14 cutaneous melanomas (88%) and 11 metastatic melanomas (92%) showed positive staining for neuron specific enolase (NSE), while only four mucosal melanomas (33%) were NSE-positive. Of the 40 melanomas, all but two were reactive with either P-S100, MoAb-079, or HMB-45. These findings suggest that MoAb-079 and NSE may be less sensitive markers than P-S100 and HMB-45 for routinely processed mucosal melanomas as compared with cutaneous and metastatic tumors.     

Immunocytochemical criteria in the differential diagnosis of malignant melanoma versus carcinoma, lymphoma and sarcoma in fine needle aspirates.

Monoclonal antibodies to keratin, vimentin, leukocyte common antigen (LCA) and S-100 protein have been used in fine needle aspirates of 35 metastatic malignant melanomas, 136 carcinomas, 35 sarcomas and 82 non-Hodgkin's lymphomas in search for immunocytochemical criteria useful in differential diagnosis of melanoma versus carcinoma, non-Hodgkin's lymphoma and sarcoma. All melanomas expressed vimentin and did not express keratin. Six of 14 melanomas contained S-100 protein. All carcinomas were keratin positive. Some were also vimentin positive. All sarcomas expressed vimentin. Synovial sarcomas were also keratin positive. All NHLs were vimentin positive, keratin negative. All NHLs except one expressed also LCA. It is concluded that keratin, vimentin and LCA are useful markers in differential diagnosis of malignant melanoma versus carcinoma and non-Hodgkin's lymphoma in fine needle aspirates when used together with morphologic and clinical data. However, in differential diagnosis of malignant melanoma and sarcoma these markers are of little use.     

婴儿色素性神经外胚瘤1例报道并文献复习

目的 探讨婴儿色素性神经外胚瘤的临床病理特征、免疫组化、诊断和鉴别诊断要点。方法 对1例婴儿色素性神经外胚瘤进行组织学和免疫组化观察和文献复习。结果 婴儿色素性神经外胚瘤好发于1岁以内的婴儿,肿瘤多见于上颌骨和颅骨,表现为浸润性和溶骨性破坏。组织学上显示大的并含不等量色素颗粒的上皮样细胞和小的神经母细胞样细胞。免疫组化显示CK、HMB-45、S-100蛋白、NSE在上皮样细胞呈阳性表达,小圆形瘤细胞S-100蛋白、NSE阳性或部分阳性。肿瘤彻底切除,随访3年未发现转移和复发。结论 婴儿色素性神经外胚瘤是一种少见的起源于神经嵴细胞的肿瘤,具有特征性的临床病理改变,需要和神经母细胞瘤、恶性黑色素瘤及其它小圆细胞肿瘤鉴别,生物学行为属于潜在恶性或低度恶性肿瘤,彻底切除预后良好。     

S100 protein, neurone specific enolase, and nuclear DNA content in Spitz naevus

The differentiation between Spitz naevus and melanoma is at times difficult. The present study was undertaken to define means to positively identify such melanocytic tumours of doubtful malignancy. Immunohistochemical staining intensity for S100 protein and neurone specific enolase (NSE) was measured in sections of 35 Spitz naevi using a microcomputer image analysis system. The data were compared with results previously obtained from 19 cases of malignant melanoma and 16 benign compound naevi. Disaggregated cells from paraffin-embedded material were stained by the Feulgen technique for DNA estimation. The nuclear DNA content distributions were measured using the same image analysis system. Compared with the malignant cases, the Spitz naevi showed significantly lower staining intensity for both S100 protein (P less than 0.0001) and NSE (P less than 0.0001). When compared with the benign compound naevi, the staining intensity was significantly lower for S100 protein (P = 0.003). The nuclear DNA distribution in Spitz naevi proved to be a normal diploid pattern in 31 cases. Four cases showed a small proportion of hyperdiploid nuclei. The results show that Spitz naevi can be significantly distinguished from malignant melanoma by staining intensity for S100 protein and NSE. A normal diploid DNA content distribution appears to be typical for Spitz naevi. Spitz and benign compound naevi show dissimilar expression of S100 protein which may indicate different patterns of differentiation in these two types of lesion. The image analysis equipment used in this study is accurate, simple to use, produces results rapidly, and is economic. Therefore, it is clinically practicable.     

Immunohistochemical detection of NSE and S-100 protein in the thalamic VB nucleus after ablation of somatosensory cortex in the rat

The ventrobasal (VB) nucleus has been studied after ablation of somatosensory cortex in 39 adult rats by the application of both NSE- and S-100 protein-immunoreactivity. Both NSE- and S-100 protein-immunoreactivity are confirmed in neurons and reactive astrocytes in the affected VB area and its surroundings, respectively. The NSE-immunoreactivity first starts in the affected VB at seven days postlesion and appears more active in its surrounding area at fourteen days postlesion. At the twenty-eight days, NSE-positive neurons are reduced in number and their stainability becomes weak. The time course of NSE-immunoreactivity is based on the progression of neuronal damage. And it is conceivable that the accumulation of NSE in neurons correlates with the regeneration. The S-100 protein-immunoreactivity is also first detected in the affected area at seven days postlesion and spread in its surrounding area at fourteen days postlesion. At twenty-eight days, S-100 protein-positive astrocytes are reduced in cell volume and their processes become thin. The time course of S-100 protein-immunoreactivity correlates with the degree of astrocytic hypertrophy. And the potent accumulation of S-100 protein appears after the onset of gliosis. The onset of neuronal damage and the repair process can be followed with immunohistochemical technique for both NSE and S-100 protein morphologically. Namely, NSE and S-100 protein can be of potential use as markers for destructive processes in the CNS.     

Immunohistochemical observation of S-100 protein and neuron specific enolase in the tumour cells of granular cell tumour

An immunohistochemical technique for the detection of S-100 protein, neuron specific enolase (NSE), carcinoembryonic antigen (CEA) and muramidase (lysozyme) was applied to a case of the granular cell tumour. S-100 protein was detected both in the nuclei and cytoplasma of the granular cells, and NSE was weakly positive in their cytoplasms. CEA and lysozyme were negative in the tumour cells. Our results supports the concept that granular cell tumours are derived from Schwann cells.     

S-100 protein and neuron specific enolase immunoreactivity of normal, hyperplastic and neoplastic chondrocytes in relation to the composition of the extracellular matrix

S-100 protein and neuron specific enolase (NSE) are no longer considered as specific cell markers indicating a neural origin. Since most of the cells displaying immunoreactivity for both markers also elaborate a stroma rich in chondroid or myxoid mucosubstances, we undertook the present study in order to clarify whether or not the positive immunoreaction is related to the composition of stromal glycosaminoglycans. The study was based on formalin fixed, paraffin embedded material comprising adult resting cartilage, reactive or hyperplastic cartilage, as well as benign and malignant chondroblastic tumors. Histochemical and immunohistochemical methods were applied on parallel sections with the following results: A positive immunoreactivity of the cartilage cells was always found to be related to the participation of chondroitine sulfate A and C in the stromal glycosaminoglycans. A NSE positive reaction was found in all cartilage cells displaying the characteristics of metabolically active cells. It is postulated that S-100 protein, as a calcium binding protein, might be involved in the cellular control mechanisms regulating the glycosaminoglycans-collagen interactions.