Didi de Wolff-Rouendaal

Monosomy of chromosome 3 and an inflammatory phenotype occur together in uveal melanoma.

PURPOSE In uveal melanoma, different predictors of poor prognosis have been identified, including monosomy of chromosome 3, HLA expression, and the presence of infiltrating leukocytes and macrophages. Each of these parameters can be used to differentiate prognostically the favorable tumors from the unfavorable ones, and thus the hypothesis for the present study was that they are related, and that monosomy of chromosome 3 occurs in the same tumors as the unfavorable inflammatory phenotype. METHODS Tumor tissue was obtained from 50 cases of uveal melanoma treated between 1999 and 2004. After enucleation, nuclei were isolated from paraffin-embedded tissue for fluorescence in situ hybridization, to determine the chromosome 3 copy number. Each tumor-containing globe was further processed for conventional histopathologic examination and for immunohistochemical analysis with HLA class I and II-specific antibodies and with macrophage marker CD68. RESULTS Of 50 uveal melanomas, 62% (31/50) were categorized as having monosomy of chromosome 3. Monosomy 3 was associated with the presence of epithelioid cells, an increased density of tumor-infiltrating macrophages, and a higher HLA class I and II expression. Survival analyses showed a correlation between monosomy 3 and decreased survival and identified monosomy 3, ciliary body involvement, and largest basal tumor diameter as the best prognostic markers. CONCLUSIONS Monosomy 3 in uveal melanoma is associated with the presence of an inflammatory phenotype, consisting of a high HLA class I and II expression as well as an increased number of tumor-infiltrating macrophages. In a multivariate Cox regression analysis, the presence of monosomy 3 was one of the best prognostic markers of metastatic disease and survival, although the follow-up time was short.

Tumor destruction by intermediate level hyperthermia

The tumoricidal effect of hyperthermia was studied in Greenes amelanotic hamster melanoma transplanted into the anterior chamber of rabbit eyes. To achieve optimal depth penetration, hyperthermia was induced with near infrared light of 820-870 nm, during 15 minutes, at a beam diameter of 2.5-6.0 mm resulting in an intermediate level hyperthermia of 45-60 degrees C. At 45 degrees C no tumor destruction occurred, at 50 degrees C the effect varied from no destruction to total thickness tumor destruction. At 55-60 degrees C total tumor destruction with additional lens damage occurred. In comparison photocoagulation with white light revealed only necrosis up to half the tumor thickness.

Retinal vasculopathy with cerebral leukoencephalopathy and systemic manifestations

Cerebroretinal vasculopathy, hereditary vascular retinopathy, and hereditary endotheliopathy, retinopathy, nephropathy and stroke are neurovascular syndromes initially described as distinct entities. Recently they were shown to be one disease caused by C-terminal frame-shift mutations in TREX1 , which was termed ‘retinal vasculopathy with cerebral leukodystrophy’. Here we defined the genetic and clinicopathologic spectrum of this clinically and pathophysiologically poorly characterized and frequently misdiagnosed fatal neurovascular disorder. We identified five different TREX1 mutations in 78 members from 11 unrelated families and by using a standardized study protocol we retrospectively reviewed and aggregated the associated clinical, neuroimaging, and pathology data. Findings were similar across mutations and families. Sixty-four mutation carriers had vascular retinopathy. Neuroimaging revealed (i) punctate, hyperintense, white matter lesions with or without nodular enhancement in 97% of them; (ii) rim-enhancing mass lesions in 84%; and (iii) calcifications in the white matter in 52%. Ninety per cent had clinical manifestations of brain disease, including focal neurological deficits (68%), migraine (59%), cognitive impairment (56%), psychiatric disturbances (42%), and seizures (17%). One mutation carrier had enhancing brain lesions and neurological features but unknown retinopathy status. Additional systemic features included liver disease (78%), anaemia (74%), nephropathy (61%), hypertension (60%), mild Raynaud’s phenomenon (40%), and gastro-intestinal bleeding (27%). Mean (± standard deviation) age at diagnosis was 42.9 ± 8.3 years and at death 53.1 ± 9.6 years. Pathological examination revealed systemic vasculopathy with luminal narrowing and multi-laminated basement membranes. The 13 mutation carriers without retinopathy or brain lesions were on average 8 years younger (mean age: 35.1 ± 10.6 years). Of them, 54% had mild Raynaud’s phenomenon, 42% had migraine, and 23% had psychiatric disturbances. Retinal vasculopathy with cerebral leukodystrophy is an autosomal dominant systemic small-vessel disease due to specific TREX1 mutations and clinically primarily characterized by (i) visual impairment from vascular retinopathy; and (ii) neurological decline and premature death due to progressive enhancing cerebral white matter lesions. Impaired liver and kidney function, anaemia sometimes associated with gastrointestinal bleeding, hypertension, migraine, and Raynaud’s phenomenon appear to be part of the clinical spectrum as well. Penetrance seems high. Because of the pathogenetic basis and the emerging clinical picture with systemic manifestations and conspicuous absence of leukodystrophy, we renamed the disease ‘retinal vasculopathy with cerebral leukoencephalopathy and systemic manifestations’. We propose diagnostic criteria to facilitate clinical recognition and future studies. * Abbreviations : AGS : Aicardi-Goutieres syndrome MC : mutation carrier RVCL(-S) : retinal vasculopathy with cerebral leukoencephalopathy (and systemic manifestations)

Decreased Expression of HLA Class II Antigens on Human Uveal Melanoma Cells After In Vivo X-ray Irradiation

In order to determine the presence of HLA antigens on human uveal melanomas, we tested anti-HLA monoclonal antibodies on tissue sections of these tumors. A great variety in expression of HLA class I and II antigens was present. A significantly lower expression of HLA class II antigens was present on uveal melanomas that had been irradiated before enucleation. These tumors lacked a lymphocytic infiltrate in comparison with nonirradiated tumors. These data suggest that radiotherapy affects expression of histocompatibility antigens on tumors.

Differential expression of HLA-A and B-alleles on uveal melanoma as determined by immuno-histology

HLA molecules play an important role in the presentation of antigens to the immune system, including tumor-specific antigens. Uveal melanomas vary in the level of expression of monomorphic HLA molecules. However, since the HLA system is polymorphic and since antigen-presentation may be linked to the expression of specific HLA alleles, the authors wondered whether allelic differences in expression existed on uveal melanomas. In order to test this, tissue sections from 23 uveal melanomas were stained in an indirect immunoperoxidase technique with monoclonal antibodies against monomorphic and polymorphic determinants of HLA molecules. All uveal melanomas showed a high level of expression of the monomorphic determinants of HLA-Class I. The polymorphic HLA-Class I molecules A2, A3, Bw4 and Bw6 varied in expression, with a higher expression of HLA-A than of HLA-B. A low level of expression of both ß2-microglobulin and HLA-B locus products was associated with a large tumor diameter. Expression of HLA-Class II molecules was low (0 to 35%). The observation that expression of the HLA-A allelic products was higher than of the HLA-B subtypes may have implications for the search of tumorspecific peptides for immunotherapeutic use: it may be worthwile to select peptides that specifically bind to HLA-A and not to HLA-B.

Hamster Greene Melanoma Implanted in the Anterior Chamber of a Rabbit Eye: A Reliable Tumor Model?

The hamster Greene melanoma (HGM), implanted into the anterior chamber (AC) of a rabbit eye, has previously been used as a model for testing experimental therapies against human uveal melanomas. Even without therapy, the tumor showed necrosis and hemorrhages 8-10 days after inoculation. These changes could interfere with the interpretation of the results of experimental therapies. In 8 rabbits, a piece of HGM was implanted subcutaneously, and after 4 weeks, HGM was also implanted in the AC of the eye. In these eyes, tumor growth in the AC slowed down, and more necrosis and hemorrhages were found clinically and histologically as compared to 8 rabbits without previous subcutaneous implantation of HGM. In spite of the long use of this tumor and frequent transfer, the tumor cells did not lose their antigenic potential.

Clinical course and pathologic features of conjunctival non-Hodgkin's lymphoma. A report of six cases

Six patients with conjunctival non-Hodgkins lymphoma (NHL) were evaluated. In all six cases the conjunctiva was the primary site of clinical presentation; four cases involved low-grade malignancy and two, intermediate-grade malignancy, according to the International Working Formulation. The disease developed within a short period of time to stage IV The poor response to therapy revealed a worse prognosis than would be expected according to the histopathologic classification. Four of the six patients died of NHL, three of them within 15 months. Comparison of these 6 cases with a group of 15 patients with orbital NHL revealed a much better prognosis in the latter patients. In spite of the fact that 4 of the 15 orbital cases showed NHL of high-grade malignancy, only 2 of them died of the lymphoma. Therefore, conjunctival NHL requires a fast and adequate diagnostic and therapeutic approach.

Effect of beta-irradiation by a 106 ruthenium plaque on the rabbit eye choroid

Fourteen Chinchilla gray rabbit eyes were treated with a ruthenium plaque, receiving 200, 400, or 800 Gy, to assess the effect of beta-irradiation on the normal rabbit choroid. Radiation effects were evaluated using fundus photography, fluorescein angiography, fluorophotometry, and histology. Fluorophotometry showed a fluorescein leakage into the vitreous 1 day after irradiation. Leakage values returned to normal within 1 month after irradiation. Fluorescein angiography showed nonperfusion of the choroid after beta-irradiation; the time between irradiation and the onset of nonperfusion was found to be dose dependent. Five months after 200 Gy irradiation, choroidal atrophy had developed but some vessels still stained with fluorescein; 400 Gy irradiation induced subtotal choroidal nonperfusion within 3 months and 800 Gy within 1 week.

Hamster Greene melanoma in the rabbit eye: immunosuppressive treatment to improve this tumor model.

Abstract• Background: The hamster Greene melanoma (HGM) implanted in the anterior chamber of the rabbit eye has been used to study experimental therapies for human uveal melanoma. However, the occurrence of spontaneous necrosis limits the value of this model for long-term evaluation of experimental treatments. In the present study we tested the hypothesis that an immune response is the cause of this necrosis and that prevention of the immune response can prolong the experimentation time • Methods: HGM was implanted in the anterior chamber of control, presensitized and immunosuppressed rabbits. The effects of sensitization and immunosuppression were assessed by clinical and histological observation • Results: Sensitization led to a significant slowing down of tumor growth, but not to a difference in necrosis. Immunosuppressive treatment with cyclosporin A improved the success rate of implantation and decreased the amount of necrosis in the tumor • Conclusion: The immune response plays a role in the occurrence of necrosis. However, although immunosuppressive treatment with cyclosporin A decreased the amount of necrosis, significant necrosis still occurred, suggesting that other factors like angiogenesis play a part as well and still limit the usefulness of this model in the long-term evaluation of experimental therapies.

Squamous cell carcinoma of the lacrimal caruncle : case reports

Purpose To report 2 cases of squamous cell carcinoma of the lacrimal caruncle. Methods Two patients, a 38-year-old man and a 72-year-old woman, presented with a painful mass in the medial angle of the eyelid aperture, with signs of inflammation. Biopsy was performed in both cases. Results Pathologic examination revealed a keratinized squamous cell carcinoma of the lacrimal caruncle in both cases. Conclusions We report 2 more cases of the rarely found squamous cell carcinoma of the lacrimal caruncle.