Prognostic immunohistochemical markers of primary human melanomas.

BACKGROUND: Several clinical and histological factors of primary melanomas comprise a relatively large quantity of prognostic information. OBJECTIVE: To find immunohistochemical markers that can improve the prognostic accuracy achieved by factors that are available without extra laboratory work, i.e. mitotic rate, tumour thickness, ulceration, localization, gender and age. METHODS: Immunohistochemical markers were determined on frozen sections. Univariate and multivariate Cox regression analyses were performed after 5-10 years follow-up. RESULTS: Seven immunohistochemical markers were related to disease-free and overall survival in univariate Cox regression analysis: Ki-67, human leucocyte antigen (HLA) -DQ, HLA-DP, Muc 18, A-10-33, transferrin receptor, and H-2-8-10. Only Ki-67 (n = 399) and HLA-DQ (n = 452) retained prognostic significance when evaluated in multivariate analyses in several models together with tumour thickness alone and with tumour thickness, gender, mitotic rate, age, localization and ulceration. CONCLUSIONS: Ki-67 and HLA-DQ may be useful for risk assessments in primary melanomas.

Can immunohistochemical markers and mitotic rate improve prognostic precision in patients with primary melanoma?

BACKGROUND: In addition to tumor thickness, several other prognostic parameters have been identified in primary human melanomas. Some are available readily (localization, gender, age, and ulceration). Others must be evaluated with a moderate or even substantial amount of work (mitoses and immunohistochemical markers). This study was undertaken to determine whether this extra effort is justified because it actually improves the precision of prognostic statements. METHODS: Immunohistologic markers were determined on frozen sections from 691 biopsies of human melanomas with the immunoperoxidase method. Univariate and multivariate Cox regression analyses were performed with metastases and with death as endpoints. RESULTS: Fifteen parameters were related to disease free survival in univariate Cox regression analysis: tumor thickness, ulceration, localization, gender, age, mitoses, and the immunohistochemical markers very late antigen (VLA)-2, human leukocyte antigen (HLA)-ABC, HLA-DR, NKI-beteb, Mel 14, intercellular adhesion molecule (ICAM-1), K-1-2, G-7-E2, and H-2-4-7. Three of the easily available parameters exhibited independent significance in multivariate Cox regression analysis: tumor thickness, ulceration, and localization. If mitotic rate was included in this model, then it had independent prognostic significance but ulceration was no longer significant. However, the model that included tumor thickness, localization, and ulceration had a slightly higher overall chi-square test score, indicating a better performance compared with thickness, localization, and mitoses. The model that included tumor thickness, localization, and mitoses could not be improved by any of the immunohistochemical markers in this study. CONCLUSIONS: Nine immunohistochemical markers with established prognostic significance for primary human melanoma were not found to improve a prognostic model that included tumor thickness, localization, and mitoses. If mitoses was replaced by ulceration, then the model performed slightly better, although ulceration was not significant in the presence of mitoses.

Amelanotic malignant melanoma presenting as malignant schwannoma.

A 78-year-old woman presented with a 14-month history of a nodule on the sole of her left foot. It had been increasing in size, and had become ulcerated. Histological, immunochemical and ultrastructural studies of the primary tumour revealed melanocytic and Schwannian characteristics, and posed diagnostic difficulties. The final diagnosis of a malignant melanoma with Schwannian differentiation was established on the basis of the clinical course, with the development of metastases in the subcutis, lymph nodes, liver and brain, as well as a shift in differentiation of the metastases towards cells containing giant melanosomes, typical of melanoma.

Tumour progression and metastatic behaviour in vivo correlates with integrin expression on melanocytic tumours.

In order to evaluate the significance of adhesion molecules expressed on melanocytic tumours for progression and prognosis in vivo, we studied integrin expression (VLA-1 to VLA-6, CD18, CD51, CD61) on 10 naevi, 40 primary malignant melanomas, and 11 metastases by immunohistology using the APAAP technique. Evaluation was done by grouping the percentage of positive tumour cells in six categories. Statistical analysis (Wilcoxon rank test, Scheffe test) revealed significant differences in the expression of VLA-1 (P < 0.0001), VLA-2 (P = 0.0001), VLA-5 (P = 0.0093), VLA-6 (P = 0.0232), and CD61 (P = 0.0002) between naevi and primary melanomas. Comparing primary melanomas with metastases, a statistically significant decrease in the expression of VLA-1, VLA-2, and VLA-6 was detectable, as well as a significant increase in VLA-4 and VLA-5. There was no correlation between integrin expression and tumour type (superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma), regression and ulceration. Changes of VLA-1, VLA-4, and VLA-6 expression correlated with the tumour thickness of the primary melanoma, but only VLA-4 and VLA-6 expression on primary melanomas correlated significantly with the development of metastases (P = 0.024 and P = 0.001). These changes of integrin expression during tumour progression particularly, the data showing an increase of VLA-4, and a decrease of VLA-6 expression support the concept that integrins are a new additional set of prognostic markers which indicate predisposition to the development of metastases.

Primary leiomyosarcoma of the skin. A histological and immunohistochemical analysis.

We report a rare case of a cutaneous leiomyosarcoma on the extensor surface of the proximal part of the left arm of an 84-year-old man. The tumour exhibited a number of histological and immunohistochemical features which are characteristic of a leiomyosarcoma. Leiomyosarcomas are frequently misdiagnosed on clinical grounds. Therefore, clinical features, differential diagnosis, histological and immunohistochemical criteria, therapy and prognosis of this rare malignant tumour are summarized on the basis of the present case and previously published reports.

Fibroblast-Derived Factors Induce Different Mast Cell Characteristics in Human Myeloid Cell Lines and Peripheral Monocytes.

Mast cell differentiation markers were studied by culturing monocytic (U937, THP1) a promyelotic (HL60) and a human mast cell line (HMC1) and peripheral human blood monocytes with fibroblast supernatants and feeder layers. Mast cell differentiation occurred on the basis of metachromasia, chloroacetate esterase, expression of mast cell-specific antigens (APAAP) and production of tryptase in all cell types, but to varying degrees and not at all with a FcεRI marker in monocytic lines. Direct cellular contact is thus not necessary for induction of mast cell differentiation by fibroblasts.

The relationship between characteristics of the tumor cells and sex of the patients in primary malignant melanomas.

Sex is a well proven prognosticator in primary malignant melanoma. We studied 19 parameters of tumor cells in primary malignant melanoma from 391 patients, the purpose being to determine if melanomas in men and women are alike. Apart from results in thin melanomas, no difference was seen between the sex groups. Thin melanomas showed a significant difference for the melanoma-associated antigen G7-E2 (higher expression in women) and the histocompatibility antigen HLA-DR (higher expression in men). The results suggest that sex-related prognostic differences are due more to host than to tumor characteristics.

p-glycoprotein expression in malignant melanoma.

In some human malignancies resistance to chemotherapy is caused by an energy-dependent efflux system, responsible for the removal of chemotherapeutics out of the resistant tumor cells. A major component of this efflux system is the permeability glycoprotein (p-glycoprotein), which depends on the multidrug-resistance gene MDR1. We have tested p-glycoprotein in primary and metastatic human melanoma by use of the monoclonal antibody C219; a substantial expression was only observed in 1/37 primary melanomas and in 1/27 melanoma metastases. None of the patients with negative metastases responded to chemotherapy. Moreover a complete remission of metastatic growth was observed in the patient with the metastasis significantly expressing the p-glycoprotein. Sequential studies revealed no significant increase of p-glycoprotein-positive cells during and after chemotherapy. We conclude that drug resistance in human melanoma does not usually depend on the p-glycoprotein-related efflux system. Other mechanisms are obviously responsible for drug resistance in this human malignancy.

Metastatic human melanoma. Phenotypic heterogeneity and antigen expression in relation to the clinical status.

According to animal experiments, metastasis to a particular organ depends on the phenotype of the tumor cells. Widespread metastatic dissemination including internal organs would therefore, at least in part, depend on the capacity of tumor cells to modulate, resulting in increased phenotypic heterogeneity. We found evidence for this assumption in human melanoma by phenotyping metastases (mainly cutaneous/subcutaneous) from 59 patients by the use of six monoclonal antibodies. Interlesional antigenic heterogeneity was present in 22/33 (67%) patients with disseminated metastases including at least one internal organ, but only in 4/26 (15%) patients whose metastases were restricted to skin and/or skin-draining lymph nodes (P less than or equal to 0.01). Chemotherapy cannot be the main reason for interlesional phenotypic heterogeneity, as seen by comparison of treated and untreated patients. Aneuploid melanoma metastases, as an indication for instability on the chromosomal level, were found in the majority of patients (84%) regardless of their clinical situation. Widespread disease was significantly related to the loss of the cytoplasmatic antigen K-1-2 and to the expression of the 130-kDa membrane antigen A-1-43.

The Ki-67 antigen in primary human melanomas--its relationship to mitotic rate and tumor thickness and its stability.

We investigated whether two parameters of proliferative activity - mitotic rate and Ki-67 positive cells - are interchangeable. The mitotic rate was assessed on paraffin-embedded sections, Ki-67 positive cells were immunohistologically determined in frozen tissue. A poor correlation (correlation coefficient r = 0.57) was found between both parameters. The proliferative activity was often not homogeneously distributed in the tested tumors. However, this is a major reason for the observed difference only in thin melanomas (less than 1.5 mm) as seen by comparison of tumors with homogeneous and inhomogeneous proliferative activity. We assume that arrest of cells in different stages of the cell cycle - variable from melanoma to melanoma - is the major reason for the observed discrepancy between mitotic rate and Ki-67 positive cells in tumors of 1.5 mm and thicker. The mean number of Ki-67 positive cells increased with tumor thickness. The stability of the Ki-67 antigen towards freezing, thawing, and formalin was studied.

[Expansion behavior of intravitreous sulfur hexafluoride in exposure to nitrous oxide]. / Zum Expansionsverhalten von intravitrealem Schwefelhexafluorid unter Lachgasexposition.

In experiments with 19 rabbits the changes in volume and concentration of intravitreously instilled sulfur hexafluoride were investigated under normal air conditions (10 rabbits in group A) and with application of laughing, gas to the inspiration system for 3 hours (9 rabbits in group B). Under normal air conditions the SF6 gas bubble attained its greatest volume after 24 hours (twice its initial volume). With the addition of 67% laughing gas to the inspiration system the largest expansion in volume (260%) was found after 3 hours, i.e., at the end of application of laughing gas, and was followed by another peak after 24 hours. With regard to the sulfur hexafluoride concentration in the gas bubble, the results in the two groups were similar. The authors conclude that in retinal surgery with application of sulfur hexafluoride general anesthesia should not include laughing gas.