Perilesional injection of r-GM-CSF in patients with cutaneous melanoma metastases.

Based on evidence that granulocyte-macrophage colony stimulating factor (GM-CSF) induces a potent systemic antitumor immunity, we tested recombinant GM-CSF in advanced melanoma. Seven patients with histologically confirmed cutaneous melanoma metastases were treated with perilesional intracutaneous injections of recombinant GM-CSF and observed for a follow-up time of 5 y. All but two patients had a decrease in the total number of metastases. At the end of the 5 y follow-up three of the seven patients are still alive with only one patient receiving other than surgical therapy, and one patient died tumor free at the age of 93. The remaining three patients died from progressive melanoma. Perilesional intradermal GM-CSF therapy resulted in a mean survival time of 33 mo. The treatment was well tolerated and no side-effects other than local erythema at the injection sites and mild drowsiness were seen. Immunohistochemical analysis with staining for CD14 and GM-CSF receptor demonstrated an increased infiltration of monocytes into both injected and noninjected cutaneous melanoma metastases compared with lesions excised prior to the initiation of therapy. The same was true for CD4- and CD8-positive lymphocytes. This phenomenon, together with GM-CSF-induced leukocyte counts of more than 20,000 during therapy, support the possible impact of a systemic over a locally induced reaction by GM-CSF. To our knowledge this is the first report that intracutaneously injected GM-CSF results in long-lasting reduction of melanoma metastases.

Low intensity laser irradiation in the treatment of recalcitrant radiation ulcers in patients with breast cancer--long-term results of 3 cases.

Radiotherapy can be followed by recalcitrant skin ulcers. As low intensity laser irradiation has been demonstrated to have a beneficial effect on impaired wound healing, we investigated its efficacy and safety in three patients with chronic radiation ulcers. The three patients, previously mastectomized due to breast cancer, with recalcitrant radiation ulcers of the skin were treated with a 30 mW helium-neon laser (wavelength: 632.8 nm, intensity: 3 mW/cm2, dose: 30 J/ cm2) three times weekly. In all patients, complete wound closure was achieved within a period of 7, 5, and 8 weeks. One patient died 6 weeks after laser treatment due to tumor cachexia. Neither of the other patients showed recurrence of radiation ulcers or neoplasm during a follow-up of 36 months. Low intensity helium-neon laser irradiation has been shown to be effective in the induction of wound healing in radiotherapy-induced ulcers in three patients with breast cancer.

First comparative delineation of the T cell receptor repertoire in primary and multiple subsequent/coexisting metastatic melanoma sites.

At present, very little is known about the types and heterogeneity of T cell responses and immunodominant epitopes of melanoma-associated antigens at coexisting sites of primary melanoma and metastatic lesions. To address this issue, we compared the T cell receptor (TCR) gene usage, complementary-determining region 3 diversity, and melanoma-associated antigens expression patterns of primary and metastatic melanoma specimens from three patients with partially homologous HLA class-1 types. Results obtained showed an overall predominance of a very limited number of TCRV regions with AV13 and BV14 being most frequently overexpressed. Sequencing of the dominating TCR transcripts confirmed the restricted usage of certain TCR specificities and, in two of the three patients, identified several identical TCR clonotypes at more than one metastatic site. Nevertheless, we failed to detect TCR transcripts that were common to all tumor deposits in a given patient and, within the majority of coexisting metastases, tumor-infiltrating lymphocytes preferentially used individual site-specifically expanded TCR beta-chain VJ segment combinations. This occurrence of individual responses simultaneously executed at and influenced in their specificity by the different sites of tumor growth, has important implications for the type of strategies chosen in the development of efficacious vaccines for patients with metastatic melanoma.

Radiographic findings in patients with acquired immunodeficiency syndrome, pulmonary infection, and microbiologic evidence of Mycobacterium xenopi.

The authors studied radiographs and clinical histories of 29 patients with acquired immunodeficiency syndrome, symptoms of pulmonary infection, and simultaneous microbiologic evidence of Mycobacterium xenopi in the respiratory tract. The presence, nature, and distribution of radiographic abnormalities were determined and analyzed in accord with clinical information. In 26 (90%) patients, M. xenopi was the only microorganism that could be isolated. Chest radiographs were normal in 13 patients (45%) and abnormal in 16 patients (55%). Radiographic abnormalities were bilateral in 94% of cases and predominantly involved the lower lobes. Patchy peribronchial opacities (44%) and miliary nodules (24%) were the most common abnormalities. Reticular opacities and parenchymal consolidation were seen in 12% of patients. Pleural effusion was seen in 18% of patients. No patients had cavitations or adenopathy. There was no statistically significant difference regarding the mean age (38.7+/-7.3 years vs. 40.2+/-11.0 years), the duration of clinically evident human immunodeficiency virus infection (2.7+/-1.2 years vs. 2.8+/-1.4 years), and the mean of CD4 cell counts (50.6+/-15.3 cells/ml vs. 47.4+/-15.9 cells/ml) between the patients with and without abnormalities on chest radiographs. In patients with acquired immunodeficiency syndrome, pulmonary infection, and simultaneous microbiologic evidence of M. xenopi, chest radiographs can be normal in a substantial number of cases. When radiographic abnormalities are present, they differ from those seen in patient not infected with the human immunodeficiency virus who had pulmonary infection caused by M. xenopi and from patients with acquired immunodeficiency syndrome and pulmonary infection with nontuberculous mycobacteria other than M. xenopi. Although these findings are not specific, they may be of importance in the imaging of patients with acquired immunodeficiency syndrome, notably in areas where M. xenopi is endemic.

Estimation of the volume-weighted mean nuclear volume discriminates keratoacanthoma from squamous cell carcinoma.

Keratoacanthoma (KA) is a fairly common neoplasm that in the past has been considered by many to be benign. Keratoacanthoma is usually differentiated from squamous cell carcinoma (SCC) by histopathologic criteria. However, the cytologic features of KA and SCC are often similar. Hence, KA may be confused with SCC at the histopathologic level. Volume-weighted mean nuclear volume (Vv) is determined by a technique that permits an unbiased and efficient estimation of nuclear volumes in tissues without any assumptions regarding nuclear shape. In this study, the volume-weighted mean nuclear volume was determined in 18 KAs and 19 SCCs to investigate whether this stereologic approach may be of use in the differentiation of these two tumors. Vv was determined by computer-assisted image analysis (IBAS 20, Kontron, Germany) on Feulgen-stained sections employing stereologic estimation of the volume-weighted mean nuclear volume. The mean Vv of KA was 704.5 microm3 (SD +/-170.5), whereas SCC exhibited a significantly lower Vv of 533.9 microm3 (SD+/-164.9) (p = 0.006). The sensitivity and specificity of Vv for the discrimination between KA and SCC was 0.80 and 0.78, respectively. We found that KAs show a significantly larger Vv than SCCs and thus, the estimation of the volume-weighted mean nuclear volume may be regarded as a helpful tool for the differential diagnosis of KA and SCC.

Quantification of vascularity in nodular melanoma and Spitz's nevus.

Spitz's nevi are acquired benign melanocytic skin tumors. Usually they are differentiated from nodular melanoma by clinical and histopathological criteria. Since Spitz's nevi are one of the most common simulators of nodular melanomas their bizarre histopathology may cause diagnostic confusion and make it difficult to differentiate these two melanocytic tumors. One of the histologic features shared by Spitz's nevus and nodular melanoma is prominent vascularity. The ability of malignant melanoma to induce angiogenesis is well established whereas benign melanocytic tumors do not have a prominent overall vascularity. The purpose of this study was to find out whether the degree of vascularity of nodular melanomas differs significantly from that of benign Spitz's nevi. In this study the number of microvessels and the vessel area were determined in 23 Spitz's nevi and 16 nodular melanomas. The number of microvessels and the vessel area were determined on Ulex Europaeus agglutinin I-stained sections by computer-assisted image analysis. Two methods of measurement were used, namely systematic and selective sampling. Measurement of the whole tumor specimen (systematic sampling) revealed a vessel count of 10.83/field (SD +/-5.97) for Spitz's nevi whereas nodular melanomas exhibited a significantly lower (p=0.04) vessel count of 6.44/field (SD +/-3.85). This difference was even more pronounced when the vessel area (Spitz's nevi: 17.85x10-4mm2, SD +/-10.32; nodular melanomas: 7.88x10-4mm2, SD +/-5.23) was investigated (p < 0.001). The difference in vessel area and vessel count was insignificant for areas exhibiting the greatest vascularity (selective sampling). Measurement of vessel count and vessel area lead us to conclude that Spitz's nevi have a significantly higher vascularity than do nodular melanomas. Our results thus indicate that angiogenesis in these pigmented lesions is not correlated with malignancy.

Increased nuclear volume in metastasizing "thick" melanomas.

Tumor invasion is the most reliable prognostic factor for primary stage I melanoma. "Thick" melanomas, with a Breslow thickness of more than 4 mm, tend to have a poor prognosis. Exceptions occur: some patients have no further recurrence of tumor. In an attempt to determine prognostic markers for "thick" clinical stage I melanomas, we investigated the volume-weighted mean nuclear volume of primary melanomas with tumor invasions > or = 4.0 mm in 32 patients. Seventeen of these patients developed melanoma metastases within a follow-up period of 60 mo; 15 patients who did not developed metastases and were comparable with regard to clinical and histological criteria were selected as a comparison group. Volume-weighted mean nuclear volume (Vv) is determined by a technique that permits an unbiased, efficient, shape- and orientation-independent, 3-dimensional estimation of nuclear size in tissues. This technique has been employed successfully in the prognostic assessment of stage I and II melanomas and was recently proven to be a sensitive marker for thin, high-risk melanomas. In our patients, Vv was determined by computer-assisted image analysis on Feulgen-stained sections by stereologic estimation of the Vv. The mean Vv (+/-SD) of primary melanomas with subsequent metastatic course was 794.99 +/- 209.18 micron3 (range: 409.48-1161.9 micron3), whereas primary melanoma lesions without subsequent metastases exhibited a mean Vv 640.54 +/- 205.07 micron3 (range: 206.7-927.48 micron3). This difference was found to be statistically significant (p = 0.0439). "Thick" melanomas with subsequent metastases thus exhibited a significantly higher Vv than did melanomas that did not metastasize.

Estimation of the volume-weighted mean nuclear volume discriminates Spitz's nevi from nodular malignant melanomas.

BACKGROUND: Spitz's nevi are benign melanocytic skin tumors that are usually differentiated from nodular malignant melanomas by histopathologic criteria. Often, however, the architectural pattern and cytologic features of Spitz's nevi and nodular melanomas are similar. Hence, Spitz's nevi may be confused with nodular malignant melanomas at the histopathologic level. EXPERIMENTAL DESIGN: The determination of volume-weighted mean nuclear volume (Vv) uses a technique that permits an unbiased and efficient estimation of nuclear volumes in tissues. In this study, Vv was determined in 13 Spitz's nevi and 14 nodular malignant melanomas to investigate whether this stereologic approach may be of use in the differentiation of these two tumors. Vv was determined by computer-assisted image analysis (IBAS 20, Kontron, Germany) on Feulgen-stained sections using stereologic estimation of the Vv. RESULTS: The Vv (+/- SD) of Spitz's nevi was 491.6 micron3 (SD +/- 175.1), whereas nodular malignant melanomas exhibit a significantly higher (p < 0.001) Vv of 775.2 micron3 (SD +/- 205.4). This difference was even more pronounced when the deeper portions of the lesions (Spitz's nevi: 443.1 micron3, SD +/- 142.4; nodular malignant melanomas: 864.1, SD +/- 169.6) were investigated. In addition, we found that in relation to the depth of the lesions the mean Vv decreased in Spitz's nevi, whereas it increased in nodular melanomas. CONCLUSIONS: We found that (i) nodular malignant melanomas reveal a larger Vv than Spitz's nevi in general, and (ii) in contrast to malignant melanomas, the Vv of nevomelanocytes in Spitz's nevi decreases in the deeper portions of the dermis. Thus, Vv may be regarded as a helpful tool for the differential diagnosis of Spitz's nevi and nodular malignant melanomas.