恶性黑色素瘤的免疫组织化学鉴别诊断

以Ｓ－１００、Ｖｉｍｅｎｔｉｎ、ＨＭＢ４５、Ｋｅｒａｔｉｎ、ＥＭＡ、ＬＣＡ抗体，ＳＰ法对原病理诊断或疑诊的３５例恶性黑色素瘤（ＭＭ）进行染色。结果１０例典型的少色素性ＭＭ均呈Ｓ－１００、Ｖｉｍｅｎｔｉｎ和ＨＭＢ４５阳性，符合原ＭＭ诊断。原病理诊断１９例和疑诊６例共２５例无色素性恶性黑色素瘤（ＡＭＭ）中，２１例同时呈Ｓ－１００、ｖｉｍｅｎｔｉｎ和ＨＭＢ４５阳性，故确诊为ＡＭＭ；４例Ｓ－１００和ＨＭＢ４５阴性肿瘤中，３例为Ｋｅｒａｔｉｎ阳性、ＥＭＡ弱阳性，１例为ＬＣＡ和Ｖｉｍｅｎｔｉｎ阳性，证实不是ＭＭ而是癌和淋巴瘤。结果表明，Ｓ－１００和ＨＭＢ４５是ＭＭ诊断性标志物，后者具有特异性。     

Immunohistochemical study of malignant fibrous histiocytoma

Malignant fibrous histiocytomas (MFH) and other soft-tissue tumors were examined immunochemically (by the peroxidase-antiperoxidase technique) for the presence and location of alpha 1-antitrypsin (alpha 1-AT) and alpha 1-antichymotrypsin (alpha 1-ACT). alpha 1-AT and alpha 1-ACT were found in substantial amounts in 19 and 20 of the 20 MFH tumors examined, respectively. In the other soft-tissue tumors, they occurred inconsistently and in less than 50% of cells, mostly those with signs of damage. It is concluded that assays for alpha 1-AT and alpha 1-ACT may aid in the diagnosis and characterization of malignant fibrous histiocytoma.     

Immunohistochemical spectrum of malignant melanoma. The common presence of keratins

Acetone-fixed frozen sections of 15 malignant melanomas of the skin with metastases were studied immunohistochemically for the presence of different types of intermediate filament proteins, synaptophysin, muscle cell actins, and desmoplakins. One of the melanomas was a primary toe tumor, and the others mainly regional lymph node metastases. The original diagnosis of melanoma was reconfirmed in each case, and the melanoma diagnosis of the metastases was verified by S100 protein immunostaining in all cases and by a monoclonal antibody to melanoma cells (NK1C3) in 7 cases. All melanomas were prominently vimentin-positive. In 10 of 15 cases, immunoreactive keratin could be demonstrated with antibody CAM 5.2. The presence of keratins was confirmed in selected cases with three other monoclonal antibodies including AE1, PKK1, and a monoclonal antibody specific for keratin number 18. Desmoplakin, another marker of epithelial differentiation, was not found in melanoma cells. Two melanomas contained neurofilament-positive tumor cells, which were however negative for synaptophysin. Desmin, muscle actins, and glial fibrillary acidic protein were not found in the neoplastic cells. On the basis of the present results one could conclude that the protein composition of the cytoskeleton of melanomas is more complex than has been previously thought and most importantly that melanomas may contain keratins.     

Immunohistochemical study of primary gastrointestinal malignant lymphoma]

63 cases of primary gastrointestinal non-Hodgkin's malignant lymphoma (ML), including 18 cases of gastric ML, 25 cases of small intestinal ML, 12 cases of ileocaecal ML and 8 cases of large intestinal ML, were studied. Small non-cleaved cell lymphomas were most common, accounting for 25.4% which was more common in the bowel than in the stomach. In 41 cases, a panel of monoclonal antibodies including L26, UCHL-1, Leu22 and Mac387 were used on paraffin section. The result of the staining was satisfactory in 37 cases. There were 35 cases (94.6%) exhibiting B-cell phenotype and 2 cases (5.4%) exhibiting T-cell phenotype. No histiocytic type was detected. The results of this and previous studies confirm the fact that most gastrointestinal lymphomas are B-cell lymphomas.     

Immunohistochemical study of experimental malignant catarrhal fever in rabbits

Malignant catarrhal fever (MCF) is an often-fatal lymphoproliferative disease of a variety of ungulates that occurs worldwide. It is caused by either of the highly related but distinct gammaherpesviruses alcelaphine herpesvirus-1 (AlHV-1, wildebeest reservoir) or ovine herpesvirus-2 (OvHV-2, sheep reservoir). MCF in rabbits is an excellent model as it closely resembles the disease in susceptible ungulates that include cattle, deer and bison. In this study, newly available and previously characterized monoclonal antibodies specific for rabbit leucocyte differentiation molecules were used to perform a detailed immunohistochemical examination of both AlHV-1 MCF and OvHV-2 MCF in rabbits. Differences in the MCF caused by the two viruses included: less tissue necrosis and more lymphoid cell accumulations in AlHV-1 MCF compared with OvHV-2 MCF, and in particular marked tissue necrosis in the mesenteric lymph node, appendix and liver of OvHV-2-infected animals when compared with either other tissues in OvHV-2 MCF or AlHV-1 MCF lesions in any tissue. In both AlHV-1 MCF and OvHV-2 MCF, lymphoid cell accumulations in lymphoid and non-lymphoid tissues consisted mainly of T-cells with a corresponding absence of B-cells. CD8(+) T-cells accounted for a proportion of these in the non-lymphoid tissues, but there was evidence for the accumulation of an unidentified T-cell subset/subsets as well. This study extends our understanding of the mechanisms of immuno-pathogenesis of MCF.     

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.     

S100 protein immunoreactivity in poorly differentiated carcinomas. Immunohistochemical comparison with malignant melanoma

This study compares the immunoprofiles of 25 cutaneous and mucosal melanomas with those of 400 poorly differentiated carcinomas, using a polyclonal antiserum to S100 and murine monoclonal antibodies to keratin (KER) and epithelial membrane antigen (EMA). All malignant melanomas expressed S100 but lacked reactivity for KER and EMA. Conversely, all of 49 carcinomas (12%) that displayed S100 also exhibited positivity for both epithelial markers. The latter group of tumors included 17 carcinomas of the breast, 16 eccrine sweat gland carcinomas, five high-grade salivary glandular adenocarcinomas, four lung cancers, three pancreatic ductal carcinomas, two serous ovarian carcinomas, one clear cell adenocarcinoma of the bladder, and one uterine cervical squamous carcinoma. None of 32 epithelial neoplasms that lacked both KER and EMA (19 testicular seminomas, four hepatocellular carcinomas, eight adrenocortical carcinomas, and one Merkel's cell carcinoma) contained S100 protein. These results suggest that S100 protein is relatively nonspecific as a single immunodeterminant in the diagnostic separation of melanoma and anaplastic carcinoma. The concomitant use of stains for EMA and KER, however, obviates this problem. Finally, since it is somewhat restricted in distribution, S100 reactivity in a known carcinoma may be of some use in predicting possible primary sources for a metastasis of unknown origin.     

Immunohistochemical expression of nm23 in primary invasive malignant melanoma is predictive of survival outcome

The objective of this study was to determine the utility of nm23 as an immunohistochemical indicator of prognosis in a large series (157 cases) of malignant melanoma and also in two subsets within this group: stage 1 tumours, whether in radial or vertical growth phase (140 cases); and stage 1 tumours in which a vertical growth phase component was positively identified (123 cases). A secondary objective was to explore the relationship between the immunohistochemical expression of nm23 and established clinical and histological indicators of prognosis in each of these three groups. In all groups it was found that strong immunoreactivity correlated positively with survival and inversely with indicators of poor prognosis, in keeping with transfection and mRNA studies and also with many immunohistochemical studies of other tumour types. That these findings are at variance with earlier reported immunohistochemical studies of melanoma highlights the importance of large case numbers of primary invasive tumours in studies which set out to explore the relationship between immunoreactivity and survival.     

Cardiac myxomas: Immunohistochemical study of benign and malignant variants

Summary Immunohistochemical investigation of 11 cardiac myxomas (CMs) including one malignant metastasizing CM showed a co-expression of epithelial (lu-5 and CAM 5.2), mesenchymal (vimentin) and neuroendocrine antigens (neuron-specific enolase) in all tumour cells. Factor VIII was found in the endothelial cells of capillaries only. In the subendocardium of fetal heart tissue close to the foramen ovale myofibroblasts reacting with the panepithelial antibody lu-5 were detected. We conclude that CMs are neoplasms that may develop from embryonic cell remnants.     

Immunohistochemical study on differential diagnosis between NPC and malignant lymphoma

Nasopharyngeal carcinoma (NPC) and malignant lymphoma are common malignant tumors which frequently involve nasopharynx and cervical lymph nodes. Sometimes, it is difficult to distinguish poorly-differentiated NPC, especially undifferentiated NPC, from malignant lymphoma. Paraffin sections of 221 cases of poorly differentiated or undifferentiated NPC and malignant lymphomas were analysed by immunohistochemical techniques (IGSS, ABC, double stain, etc.), The immunohistochemical criteria of differential diagnosis between NPC and malignant lymphomas were proposed and with these criteria, 40 cases which were difficult to distinguish between NPC and malignant lymphoma were identified. In comparison with the methods of SPA, PAP, ABC, IGSS, etc., and the probes of Ke, EMA, LCA, Vi, etc. on paraffin sections, IGSS or ABC method and probes of Ke and LCA were considered to be more sensitive.     

Immunohistochemical and ultrastructural study of human melanoma colonies grown in soft agar

An immunohistochemical and ultrastructural study of human melanoma colonies grown in soft agar for up to 50 days was performed. Three morphological variants of developing tumor colonies are reported: (1) large light colonies, (2) small dark colonies, and (3) smooth-edged colonies. The large light colony variant is the most frequently observed in the soft agar assay (≈?70%), followed by the dark colony variant (≈?27%), and the smooth-edged colony variant (≈?3%). Major morphological characteristics are associated with each variant, as shown with light microscopy (LM) and transmission electron microscopy (TEM). Both LM and TEM analyses demonstrated that the large light colony variant was hypomelanotic and contained a microfibrillar extracellular matrix (ECM). The small dark colony variant was found to be hypermelanotic and contained a less demonstrable ECM. The smooth-edged variant has an encapsulated periphery, no demonstrable ECM, and tightly packed cells with desmosome-like junctions. In order to characterize further the ECM in the most commonly observed variant, the large light colony, specific antibodies to fibronectin (FN) and collagen types IV and V (COLs IV and V) were applied and observed with immunofluorescence microscopy and immu-noperoxidase. In paraffin sections of melanoma colonies, FN was observed associated with both the cell surface and the ECM. However, no specific staining was seen for COLs IV and V. In addition, ruthenium red was used to preserve and selectively bind to glycosaminoglycans (GAGs) and proteoglycans (PGs). TEM studies reveal GAG-like granules stained with ruthenium red in the fibrillar ECM and a dotted, punctate staining of the cell surface. Understanding the biological and architectural composition of developing melanoma tumor colonies in soft agar could contribute to the development of more efficient chemo-therapeutic strategies.     

Immunoscintigraphy of malignant melanoma

This work is part of a multicentric European evaluation of the monoclonal antibody 225.28s targeted against malignant melanoma and its metastases. Twenty-eight patients (12 males, 16 females, mean age: 53 yrs), who had initially been treated by resection of the primary tumour, were included in the study. Twenty-three of the 26 metastases more than 1 cm in diameter were visualized by immunoscintigraphy. The sensitivity of the procedure (88%) is limited however by the small size of the lesions and their depth, as well as by background noise caused by circulating antibodies. Immunoscintigraphy enables non-invasive investigation of the whole body and can detect lesions that other conventional complementary explorations fail to identify.     

Desmoplastic malignant melanoma. A clinicopathologic study of 14 cases

Clinical and pathologic details in 14 cases of desmoplastic malignant melanoma were reviewed. The study group included ten men and four women with a median age of 58 years. Anatomic locations such as the head and neck area (four cases), limbs (five cases), and trunk (five cases) were involved with equal frequency. Follow-up information (median period, 4.6 years) was available for 12 patients, of whom four are alive and disease free, six have had local tumor recurrence, and two have died of their disease. Histologically, these lesions consisted of a malignant fibroblastic skin tumor intimately associated with a superficial melanoma (ten cases) or melanocytic dysplasia (four cases) that often extended deeply to the subcutaneous fat. Helpful diagnostic features included the presence of neurotropism, a lymphocytic infiltrate, and unusual patterns of triangular and periadnexal lamellar fibroplasia. Of the immunohistochemical markers employed, antisera to S100 protein and vimentin yielded the most consistent positive results. Immunostaining with NK1/C-3 (antimelanoma monoclonal antibody) was not helpful. Ultrastructural evidence of fibroblastic and schwannian differentiation was seen. We conclude that the altered morphologic melanomas is associated with a relatively favorable prognosis and believe that careful attention to light microscopic detail with immunohistochemical and electron microscopic assistance will elucidate the diagnosis in most cases.     

Immunohistochemical expression of C-myc oncogene,heat shock protein 70 and HLA-DR molecules in malignant cutaneous melanoma

The clinical course of malignant melanomas is frequently unpredictable, although a number of prognostically useful variables can be identified. There is a need for additional markers of prognostic value. In a series of 60 malignant cutaneous melanomas, we analysed the immunohistochemical expression of c-myc proto-oncogene, heat shock protein 70 (HSP70) and HLA-DR molecules in order to investigate their prognostic significance. C-myc, HSP70 and HLA-DR were expressed in 43.3%, 56.6% and 38.3% of all melanoma cases, respectively. Advanced Clark levels (Clark III–V) were significantly associated with c-myc expression rate (P<0.05), HSP70 detection (P<0.01) and HLA-DR positivity (P<0.01). Increased Breslow thickness (>1.5 mm) was related to HLA-DR expression (P<0.05). High mitotic rate was closely associated with c-myc positivity (P<0.05), while HSP70 and HLA-DR expression separately correlated to clinical stage of the disease (P<0.05). The evaluation of these variables may be of immunological and prognostic significance. They were found to be associated with melanocyte subpopulations of the vertical growth phase which are arguably characterized by an increased invasive potential.     

Immunohistochemical staining in malignant mesotheliomas

The immunoreactivity of five antibodies was evaluated on six routinely processed mesotheliomas to evaluate their ability to distinguish mesothelioma from metastatic adenocarcinoma. The diagnosis in all cases was confirmed by electron microscopic examination and histochemical stains for neutral mucin (periodic acid-Schiff-diastase) and acid mucin (alcian blue with and without hyaluronidase). AE1, a monoclonal antikeratin antibody that stains most carcinomas, reacted with all six cases of mesothelioma. HMFG-2 and anti-epithelial membrane antigen (antibodies reactive with human milk fat globule proteins), two other closely related antibodies reactive with most carcinomas, also reacted with all of the mesotheliomas in the authors' series. A polyclonal antibody to carcinoembryonic antigen (anti-CEA) did not stain any of the mesotheliomas in their series. Anti-Leu-M1 did not react with the mesotheliomas. The authors conclude that none of these antibodies, when used alone on routinely fixed paraffin-embedded material, is both sensitive and specific in the distinction of mesothelioma from adenocarcinoma. However, immunoperoxidase studies using anti-CEA and anti-Leu-M1 may occasionally be helpful when used in conjunction with other histochemical stains and electron microscopic examination in distinguishing mesothelioma from metastatic adenocarcinoma.     

Ovarian malignant melanoma: a clinicopathologic study of 5 cases

Primary ovarian malignant melanomas are extremely rare. The wide range of morphologic appearances assumed by melanomas in the ovary can cause considerable difficulty in diagnosis. The clinicopathologic features of 4 definite and 1 probable primary ovarian melanomas are presented. The patients ranged in age from 41 to 71 years. Four tumors were within teratomas with 2 showing a lentiginous pattern of melanoma in the squamous epithelium. Unusual histologic features were noted. Immunostains for S-100, HMB-45, and Melan-A were positive in all tumors. Premelanosomes were identified in 2 tumors ultrastructurally. Metastatic sites included regional nodes, peritoneal surfaces, omentum, lung, liver, brain, and bone. All 5 patients died within 18 months. Immunohistochemistry and electron microscopy aid considerably in the diagnosis of ovarian melanomas where pigmentation or teratomatous elements are absent. Familiarity with the wide range of morphologic patterns presented here will raise awareness and facilitate detection of future cases of ovarian melanoma.     

Cytodiagnosis of amelanotic metastatic malignant melanoma: An immunocytochemical study

Malignant melanomas are known to present diverse patterns and this can result in considerable difficulties for an interpretation of malignancy type from cytohistologic material. Such difficulties are further compounded, if melanin pigment cannot be demonstrated by conventional histochemical stains; unfortunately many cases do exhibit this feature, especially in its metastases. This study was designed to review cases of amelanotic metastatic malignant melanomas in a variety of cytologic samples which were received from 40 patients with a known history of malignant melanoma. Cytomorphologically, the cases were classified as classical, carcinoma-like, spindle cell type, lymphoma-like, and undifferentiated type (Table I). While in 36 of the cases, the diagnosis of metastases from a melanoma was confirmed based on an immunopositivity to a variety of melanoma markers (Table III), in four of the cases, the immunostaining indicated metastases from another primary source, which was subsequently found. Based on our study, we are of the opinion that an immunologic characterization is useful to conclusively diagnose the majority of cases of metastatic amelanotic malignant melanomas. Furthermore, we feel that a reliance on a single melanoma marker is not justified, and a panel of antibodies should be used to distinguish a metastatic amelanotic malignant melanoma from other metastatic neoplasms. Diagn. Cytopathol. 16:238–241, 1997. © 1997 Wiley-Liss, Inc.