Pyrogenic cytokine interleukin-6 expression by a chordoid meningioma in an adult with a systemic inflammatory syndrome. Case report and review of the literature.

Chordoid meningioma is a rare meningothelial tumor characterized by chordoma-like histological features with lymphoplasmacellular infiltration. This tumor is often seen in children, but not in adults, with a systemic inflammatory syndrome (iron-resistant microcytic anemia and/or dysgammaglobulinemia) and very rarely with a persistent moderate hyperthermia. In the present report the authors describe a temporal chordoid meningioma in a 30-year-old woman who presented with fever, headache, and a serological inflammatory syndrome. The clinical symptomatology, chiefly the fever, disappeared immediately after removal of the tumor. To the authors' knowledge, only one similar patient with such clinical presentation and response to surgery has been mentioned in the literature. Interestingly, at immunohistochemical examination, the neoplasm showed focal positivity for the pyrogenic cytokine interleukin-6. The capacity of the tumor to produce this pyrogenic cytokine could explain both the patient's clinical presentation and her response to the surgical management.

Intrasinusoidal metastatic melanoma of the liver with reactive hepatitis. An immunologic phenomenon?

The macrophage migration inhibition factor (MIF) test toward extracts of choroidal melanoma was repeated four times in a patient with ocular malignant melanoma. In the initial stage, when there was only an ocular finding, the MIF test result was positive. It remained so for a period of two years, even when intrasinusoidal hepatic diffusion developed concomitantly with a nonspecific reactive hepatitis. These histologic findings can be interpreted as evidence of the presence of an immune reaction at a particular moment in the disease process. Several months later, when the patient's condition went into an abrupt decline and showed extensive nodular spread, the results of MIF test were found to have become negative.

[Serum antiretinal antibodies (ARA) in patients with choroidal melanoma--preliminary report]. / Przeciwciala przeciwsiatkówkowe (PPS) w surowicy chorych z czerniakiem naczyniówki--doniesienie wstepne.

PURPOSE: To estimate serum antiretinal antibodies (ARA) in patients with choroidal melanoma after therapy in 3 months follow-up period. MATERIAL AND METHODS: 24 patients at the age 37-72 years (mean: 57.8 years) with choroidal melanoma were examined. In all cases routine ophthalmic examination with A- and B-scan ultrasonography and fluorescein angiography were performed. In some subjects also indocyanine green angiography was performed. Ruthenium-106 plaques were used in 16 cases, in 5 cases treatment with Iodine-125 plaques was performed. Three patients underwent enucleation. In all cases ARA were determined in a serum dilution 1:10 by indirect immunofluorescence test on normal monkey retina as a substrate and FITC-labeled anti-human IgA, IgG, IgM serum (Euroimmun--Germany). ARA were scored before and within 3 months, after the therapy was performed. RESULTS: Before treatment ARA were present in 15 patients (62.5%) with choroidal melanoma in serum dilution 1:10. In a control group ARA were present in 3 cases (12.5%) in serum dilution 1:10. After therapy within 3 months follow-up period ARA occurred in serum of further 7 patients, all treated with Ruthenium-106 plaques. Fluorescence of outer retinal segments was present in 7 cases, while serum of remaining 15 patients demonstrated positive reaction within the whole tissue. Indirect immunofluorescence test on normal monkey retina revealed in 11 patients the presence of other autoantibodies; antinuclear and antinucleolar antibodies, reacting with nuclei of inner and outer nuclear layers of retina. CONCLUSIONS: The presence of ARA in serum of patients with choroidal melanoma before therapy may be a result of autoimmune reaction against the tumor tissue. The detection of ARA in serum after plaque therapy may be associated with reaction due to tumor irradiation. The appearance of other serum autoantibodies in patients with choroidal melanoma may indicate at more advanced and generalized process.

Missing evidence for HLA antigen association with Eales' disease, chorioretinitis, central serous retinopathy, and malignant choroidal melanoma.

Patients with Eales' disease, chorioretinitis, central serous retinopathy, or malignant choroidal melanoma were tested for HLA antigen deviation. When corrected p values (pc) are used, the first three disorders did not show any significant deviation, whereas a significant increase of HLA-Aw32 (pc = 0.026) was found in the malignant melanoma group. For conclusive evidence the latter finding needs confirmation by analysis of a greater number of patients with this disorder.

Inhibition of leukocyte migration with melanoma-associated antigens in choroidal tumors.

Patients with choroidal melanomas and simulating choroidal tumors were tested in a masked manner in a leukocyte-migration inhibition (LMI) assay with a 3M KCl antigen derived from a metastatic choroidal melanoma. Good discrimination petween patient groups was observed with this antigen. The sensitivity and specificity of this assay are discussed.

Inhibition of macrophage migration by choroidal malignant melanoma-associated antigens in patients with unveal melanoma.

The specific cell-mediated immunity of the lymphocytes of eight patients with choriodal malignant melanoma (MM) to four extracts of choroidal MM-associated antigens was tested with the aid of the MIF technique. Seven of the patients with choroidal MM responded to at least one of the four extracts used, whereas patients with choroidal nevus or carcinoma as well as healthy controls did not respond to any of the MM choroidal extracts. There was no response to iris extracts obtained from the enucleated eyes with MM in any of the subjects tested.

Immunolocalization of integrins in the human retina.

PURPOSE: Integrins are cell surface proteins that participate in interactions between cells and with extracellular matrix. Binding of integrins to their ligands influences cell activities including proliferation, migration, and differentiation. Expression of integrin subunits from three different subfamilies were examined in human retina. METHODS: Integrins were detected in frozen sections of two human retinas with an avidin-biotin-complex immunohistochemical technique, using nine different monoclonal antibodies specific for alpha 2, alpha 3, alpha 4, alpha 5, alpha 6, alpha v, beta 1, beta 2, and beta 3. One retina was from a patient who had conjunctival squamous cell carcinoma, and the other was from uninvolved regions of an eye with a choroidal melanoma. RESULTS: All integrins tested were detectable in consistent patterns in two retinas. All except alpha 2 and alpha 4 were stained vibrantly in retinal and choroidal vessels. All alpha subunit staining of vessels showed overlap or close proximity to beta 1 staining. In addition to vessels, beta 1 was also present in the internal limiting membrane; alpha 2, alpha 3, alpha 4, alpha 5, and beta 2 were all found throughout much of the neural retina, albeit with distinctive staining patterns. Other than in association with vessels, alpha 6 and alpha v were not detected in neural retina, and beta 3 was only weakly detected in the nerve fiber layer; alpha 4 and beta 2 were expressed in the retinal pigment epithelium; beta 1 and beta 2 were strongly expressed in drusen present in one of the eyes. CONCLUSION: Nine integrin subunits have been found to have unique distributions in adult human retina. An understanding of the distribution in normal retina can serve as a useful contrast to patterns of staining associated with retinal diseases.

Studies of tumor-infiltrating lymphocytes from a human choroidal melanoma.

Cell suspensions prepared enzymatically from an ocular choroidal melanoma were cultured in vitro in an effort to generate (1) melanoma tumor cell lines and (2) tumor-infiltrating lymphocytes cytotoxic for ocular melanoma cells. Even though histologic study of the tumor did not show "significant" infiltrating bone marrow-derived cells, lymphocytes were generated readily in cultures to which interleukin-2 was added. Phenotypic analysis of the cultured lymphocytes indicated that T-cells, natural killer (NK) cells, and lymphokine-activated killer (LAK) cells were present. Moreover, functional studies of the cultured lymphocytes revealed NK activity, LAK activity, and most importantly, tumor antigen-specific cytotoxic T-cell activity. It was concluded that it is possible to obtain tumor cell lines and tumor-infiltrating lymphocytes from ocular tumors, both of which would be required if cellular immunotherapy of ocular tumors is contemplated. In addition, these results indicate that ocular melanomas can express unique tumor-specific antigens and that the immune system of a patient with such an ocular tumor can perceive these tumor antigens because antigen-specific precursor cytotoxic T-cells were present in the tumor-containing eye at the time of enucleation. The theoretic and therapeutic implications of these findings are discussed.

Membrane-bound regulators of complement activation in uveal melanomas. CD46, CD55, and CD59 in uveal melanomas.

PURPOSE: To identify the presence of membrane-bound regulators of complement activation (m-RCA) on uveal melanomas and uveal melanoma cell lines and to examine their role in the inhibition of complement-mediated lysis in vitro. METHODS: Immunohistochemistry and flow cytometric analysis with monoclonal antibodies directed against m-RCA CD46, CD55, and CD59 were applied to tissue sections of 10 uveal melanomas, three primary uveal melanoma cell lines, and one uveal melanoma metastatic cell line. A microcytotoxicity test was used for measuring antibody-dependent complement-mediated lysis. RESULTS: The tissue sections and all four uveal melanoma cell lines expressed CD46, CD55, and CD59. Complement-mediated lysis in the presence of human complement was increased after partial removal of the m-RCA CD55 and CD59 with phosphatidylinositol-specific phospholipase C from the uveal melanoma cell line 92-1. CONCLUSIONS: These results demonstrate that CD46, CD55, and CD59 are expressed in uveal melanomas and that CD55 or CD59, or both, plays a role in resistance to complement-mediated cytotoxicity. The finding that m-RCA are expressed in uveal melanomas may have implications for the effectiveness of the anti-tumor response and in the therapeutic application of monoclonal antibodies directed against tumor-associated antigens.

Frequent loss of heterozygosity on chromosome 6p in uveal melanoma.

Lack of expression of HLA class I antigens is frequently observed on primary uveal melanoma, and is correlated with improved patient survival. Several mechanisms may contribute to the observed loss of HLA class I expression, including changes at the DNA level. In this study, we used microsatellite analysis as a molecular genetic approach to examine loci on chromosome 6p for loss of heterozygosity (LOH). Three pairs of microsatellite markers were used to screen 20 formalin-fixed, paraffin-embedded uveal melanomas for LOH on the short arm of chromosome 6. In all cases, normal adjacent scleral tissue was used as a control. We identified LOH in eleven cases from microsatellite locus D6S105 to the telomere, in eight cases from microsatellite locus D6STNFa to the telomere (area includes D6S105), and in seven cases from microsatellite locus D6S291 to the end of chromosome 6p (includes D6STNFa and D6S105). In seven cases, retention of heterozygosity was found at all three loci using these primers. Our results suggest that loss of heterozygosity on chromosome 6p is a common feature in uveal melanoma. We did not find a correlation between the presence of LOH and locus-specific HLA-A and -B expression.

Susceptibility of human ocular melanoma cells to spontaneous and interferon-augmented natural cytotoxicity.

Due to the current interest in natural killer (NK) cells as a host defence mechanism against neoplasia, we have investigated the susceptibility of short-term cultures of human ocular melanoma cells to spontaneous and interferon (IFN)-augmented natural cytotoxicity. Cultures of ocular melanoma cells were readily established and identified as bipolar and multipolar pigmented melanoma cells. In short-term cytotoxicity assays these cell lines demonstrated a weak susceptibility to spontaneous human PBL natural cytotoxicity, and increased killing was observed using IFN-augmented cytotoxic effector cells. These findings may be pertinent in relation to the role of NK cells in vivo.

Antigenic and cellular heterogeneity of primary uveal malignant melanomas.

Monoclonal antibody (MAb) MAb8-1H, which identifies an ocular melanoma-associated antigen, and computer-assisted cytomorphometric analysis were used to investigate antigenic and cellular heterogeneity in four atypical cases of choroidal or ciliary body melanoma. The presence of one or more well-localized, morphologically distinct areas within each uveal melanoma, suggestive of multiple tumor cell populations, was observed by conventional light microscopy. In one choroidal melanoma with an unusual bilobed configuration, MAb8-1H bound only to the anterior lobe composed of spindle melanoma cells. In the other three cases, MAb8-1H bound selectively to discrete regions within the uveal melanoma corresponding to the well-localized, morphologically distinct areas seen by light microscopy. In each case, a computer was used to calculate the nuclear area of 50 tumor cells corresponding to the areas binding or not binding MAb8-1H. In three of the four cases examined, the differences in mean nuclear area between the discrete populations of tumor cells were statistically significant. Our results suggest that some uveal melanomas are composed of two or more discrete populations of tumor cells with regard to both expression of melanoma-associated antigens and cytomorphologic features.

Metastatic uveal melanoma. Hepatic metastasis identified by hybridoma-secreted monoclonal antibody MAb8-1H.

Monoclonal antibody MAb8-1H, which defines an ocular melanoma-associated antigen retained in fixed tissue, was used to demonstrate metastatic melanoma cells in liver biopsy specimens from five patients with primary choroidal melanoma. In all cases, the monoclonal antibody bound to tumor cells and not to normal hepatocytes. In one case, a well-delineated focus of tumor cells bound to MAb8-1H; in two cases, MAb8-1H appeared to delineate individual tumor cells that were difficult to categorize by conventional staining techniques. The liver biopsy specimens were almost completely replaced by tumor cells in the remaining two cases. This monoclonal antibody may be applicable to the diagnosis of metastatic uveal melanoma and to delineate micrometastasis.

Carcinoembryonic antigen. Its role in the evaluation of intraocular malignant tumors.

Plasma carcinoembryonic antigen (CEA) was studied in 60 patients with histologically confirmed intraocular neoplasms including 56 malignant melanomas of the uvea and four metastatic tumors to the choroid. While 45% of the patients with primary uveal melanomas, as well as 75% of the patients with metastatic disease demonstrated elevated plasma CEA levels, both patients who exhibited metastatic lesions of entodermal origin demonstrated plasma CEA values that clearly fell into a separate, highly elevated category, consistent with metastatic disease or pancreatic or colorectal carcinoma. Thus, in the patient seen with a nonpigmented choroidal mass that may represent either a choroidal hemangioma, amelanotic melanoma, or metastatic tumor, plasma CEA levels may be useful in the differential diagnosis. If the clinician suspects a metastatic tumor from an occult primary site, highly elevated CEA levels may indicate that the lesion is of entodermal origin.

Combined immunotherapy of malignant melanoma. Unusual survival following cerebral metastasis.

An 18-year-old woman was found to have solitary cerebral, choroidal, and pulmonary metastases of malignant melanoma three years after excision of a primary malignant melanoma. The cerebral metastasis was excised, and the patient's condition was treated with CNS irradiation followed by combined immunotherapy with transfer factor and Bacille bilié de Calmette-Guérin. The transfer factor donor was her father, who showed cellular immunity to melanoma extracts on in vitro testing. Histologic examination of the pulmonary nodule, which was excised after the initiation of immunotherapy, revealed a dense lymphocytic infiltrate associated with the metastatic melanoma. The patient is currently free of detectable melanoma more than three years after the cerebral metastasis. Studies in a second patient also demonstrated the appearance of inflammatory infiltrate in metastatic melanoma following transfer factor therapy.

Cell-mediated immunity to melanoma-associated antigens in patients with ocular malignant melanoma.

We tested 11 patients with choroidal melanomas and 32 control subjects for cell-mediated immunity to melanoma-associated antigens by an in vitro leukocyte migration inhibition assay. Five of seven patients with choroidal melanomas, who received two melanoma extracts used in this series of experiments, had significant leukocyte migration inhibition as compared with none of 17 normal subjects. Four of five melanoma patients who received a third melanoma extract had significant leukocyte migration inhibition as did three of four normal controls, indicating that the extract showed nonspecific inhibition. These data agreed with the concept that patients with choroidal melanomas have cell-mediated immunity to common melanoma-associated antigens.

Immune responsiveness in patients with choroidal malignant melanoma.

We studied selective in vivo and in vitro immunologic factors of 25 patients with choroidal malignant melanoma. Immune profile factors did not show consistent differences between patients with melanoma and an age-matched control population. The studies were also non-contributory in delineating variations in immunocompetence with respect to melanoma cell types. Serial immune profiles failed to identify a change when clinical evidence of metastasis developed. When tested as a group, patients with melanoma in whom metastatic disease developed during the study showed significant depression of initial mitogen-induced lymphocyte blastogenesis. This depression was noted in advance of clinically recognized metastatic disease. This group had a wide range of responses that overlapped those of control patients, thus making the usefulness of these testing factors questionable for individual prognosis or for clinical monitoring.

Immune complexes and carcinoembryonic antigen levels in metastatic choroidal tumors.

We studied 11 patients with metastatic choroidal tumors and one patient with a metastatic choroidal melanoma by using the Raji cell radioimmunoassay to detect immune complexes, and the carcinoembryonic assay. Ten (91%) patients with choroidal metastases had an increase of either immune complex levels or carcinoembryonic antigen results.

Flow cytometry analysis of peripheral blood lymphocytes in patients with choroidal melanoma.

PURPOSE: To evaluate peripheral blood lymphocyte subpopulations in patients with choroidal melanoma. METHODS: In this prospective study, peripheral blood lymphocytes of 226 patients afflicted with choroidal melanoma were analyzed by flow cytometry and compared with those of 49 age-matched and gender-matched control subjects. Subpopulations of peripheral blood lymphocytes were further identified by monoclonal antibodies specific to cell-surface markers. Statistical analysis was performed by a Student t test. RESULTS: There was no overall difference between the patients with choroidal melanoma and the control subjects with regard to peripheral blood lymphocyte subpopulations. However, when the patients were divided into subgroups based on their clinical characteristics, differences in natural killer (NK) cell population and activated T cells were noted in two subgroups. Patients with ciliary body involvement showed a statistically significant reduction in NK cells (194 +/- 101 vs 260 +/- 178 per mm3; P = .01). The number of activated T cells in this subgroup of patients was increased but not statistically significantly (7.32 +/- 4.79 vs 6.09 +/- 4.34 per mm3; P = .08). In patients with extrascleral extension, a statistically significant increase in activated T cells was noted (9.84 +/- 7.41 vs 6.25 +/- 4.3 per mm3; P = .02). The NK cells in this subgroup of patients were also reduced, but the reduction did not achieve statistical significance (178 +/- 123 vs 248 +/- 167 per mm3; P = .24). CONCLUSIONS: We noted statistically significant differences in peripheral blood lymphocytes in two subgroups of patients with clinically less favorable choroidal melanoma.