Establishment of a human leiomyosarcoma cell line.

A cell line designated SKN was established from the human uterine leiomyosarcoma of a 52-year-old female. The cell line has grown well and the serial passages were successively carried out 82 times within 12 months. The monolayer cultured cells revealed anaplastic and pleomorphic features, and they multipled rapidly without contact inhibition. Electron microscope studies revealed myoibrils but no virus-like particles, while chromosomal studies showed that all cultured cells were hyperploid, the modal number was 112, and the marker chromosome was present. The cells were transplanted into an immune-depressed hamster cheek pouch and produced a histological leiomyosarcoma resembling the original tumor.

Novel functions and cellular interaction of human lymphoid stromal cells with lymphoid cell lines in vitro.

Lymphoid stromal (SG) cells have been isolated from the lymph node of a patient with malignant lymphoma, and characterized by positive reaction with a monoclonal antibody against the T-zone stromal cells in human lymph nodes. B-acute lymphoblastic leukemia (BALL) cells showed prominent emperipolesis toward SG cells when they were cocultured, whereas T-acute lymphoblastic leukemia (TALL) cells attached firmly to the surface of SG cells. Autologous peripheral B and T cells behaved, respectively, in the same way as BALL and TALL cells. Both BALL and TALL cells while directly interacting with the SG cells were completely inhibited from incorporating [3H]thymidine, although radioactive grains were observed in 16.4%-12.4% of supernatant BALL and 13.8%-13.0% of supernatant TALL cells in each coculture. Furthermore the media conditioned by SG cells significantly increased the incorporation of [3H]thymidine into the TALL cells as much as 190% of the control. These results indicate that SG cells undergo tissue-specific cellular interactions with B- and T-lymphoid cell lines but not with a myeloid cell line, and they can modify their growth by two distinct mechanisms. SG cells proved to be very useful in studying the effect of the lymphoid microenvironment on the proliferation of lymphocytes in vivo.

Prognostic factors of stage I and II non-Hodgkin's lymphomas of the head and neck: the value of the working formulation and need for chemotherapy.

From 1966 through 1984, 142 patients with early stage non-Hodgkin's lymphoma of the head and neck were treated. Histologic slides were reviewed and reclassified according to Rappaport's classification and the Working Formulation. Patients were clinically staged; 82 Stage I, and 60 Stage II. Five-year freedom from relapse was 53% for Stage I and 48% for Stage II patients with no statistically significant difference. No patients with favorable histologies of Rappaport's classification or low grade malignancy of the Working Formulation relapsed. By univariate analyses, stage, primary site, the Working Formulation, Rappaport's classification, extent of radiation field, radiation dose, tumor bulkiness and addition of multiple-agent chemotherapy seemed to be prognostic factors for predicting relapse. Multivariate regression analyses (MVA) showed primary site and multiple-agent chemotherapy were independent prognostic factors. Tumor bulkiness provided marginal prognostic significance. Waldeyer's ring lymphomas fared better than nodal or extralymphatic lymphomas; it seems necessary to report Waldeyer's ring lymphomas independently from nodal or extralymphatic lymphomas. Patients receiving more than 3 chemotherapeutic agents had better prognosis than those receiving 3 or less agents, or no chemotherapy. Pathologic grade was another prognostic factor when the Working Formulation was used in MVA. When Rappaport's classification was used, division of unfavorable histologies into histiocytic and non-histiocytic groups provided only marginal significance in MVA. We conclude that Stage I-II non-Hodgkin's lymphomas of the head and neck with favorable histologies of Rappaport's classification or low grade malignancy of the Working Formulation be treated with radiation therapy only; and patients with other histologies should be treated with multiple-agent chemotherapy and radiation therapy.

Non-Hodgkin's lymphoma confined to the nasal cavity: its relationship to the polymorphic reticulosis and results of radiation therapy.

From 1975 through 1988, nine patients with locally confined nasal non-Hodgkin's lymphoma (NHL) were treated with radiation therapy in the Department of Radiology, Chiba University Hospital. Immunohistochemical study disclosed that all NHL's have T-lineage. Additionally, unique histological pictures of polymorphism, angiodestruction, and necrosis were seen in most of cases. These three findings are the histological features of polymorphic reticulosis (PMR), which is the main cause of lethal midline granuloma and has recently been shown to belong to T-cell malignancy. Therefore, it is concluded that the nasal T-cell NHL and PMR are really a single disease entity. The predominance of the T-cell lymphoma in the nasal cavity as well as its histological distinctness clearly indicate that the head and neck extranodal NHL cannot be discussed together. Although the disorder was considered to be locally limited at presentation, only 3 of the 9 patients with nasal NHL could be induced into long-term remission with involved field radiotherapy. The distant extranodal spread was the primary cause of failure. Multimodality treatment using intensive chemotherapy might improve the prognosis of nasal NHL.

Herpes simplex lymphadenitis. Report of two cases with review of the literature.

We report two cases of herpes simplex lymphadenitis without widespread organ involvement in a 60-year-old man and a 67-year-old woman. Their complaints were high fever and generalized erythema followed after few days by generalized lymphadenopathy. This report describes the findings obtained by light and electron microscopy, immunohistochemistry, and in situ hybridization. In both instances, Cowdry's type A intranuclear inclusion bodies were found in T-immunoblasts in the background of T-zone hyperplasia with focal necrosis. Electron microscopic investigation revealed intranuclear and cytoplasmic virus particles with characteristics of the herpes group. Immunohistochemical staining utilizing anti-herpes simplex virus (HSV) antibody was positive and in situ hybridization with HSV-DNA probe revealed positive signals in the nucleus and in the cytoplasm of T-immunoblasts. Although rare, HSV lymphadenitis in the absence of generalized infection can occur.

Histogenesis of alveolar soft part sarcoma. An immunohistochemical and biochemical study.

In order to clarify the histogenesis of alveolar soft part sarcoma (ASPS), an immunohistochemical and biochemical study was performed on three cases. The immunohistochemical study indicated the presence of actin, desmin, vimentin, and Z-protein in all cases. On the other hand, intermediate filaments other than desmin and vimentin were not detected immunohistochemically. The presence of desmin and Z-protein strongly suggests the myogenic character of this tumor. As to whether ASPS shows striated muscle differentiation or smooth muscle differentiation, the immunohistochemical absence of myoglobin in the three cases suggests that the tumor does not differentiate in the direction of striated muscle. However, biochemical assay of subunits of enolase revealed significantly high amounts of beta-enolase, which is known as a marker for striated muscle, in all three cases. The determined values--735, 426, and 584 ng/mg of protein --are indicative of striated muscle differentiation. In addition, the immunohistochemical study of all cases revealed the presence of beta-enolase in tumor cells. These data definitely show the myogenic character and rhabdomyoblastic differentiation of ASPS.

Solitary (localized) pleural mesothelioma. A light- and electron-microscopic study.

Six solitary (localized) pleural mesotheliomas were studied by light and electron microscopy. All the lesions were benign and were composed mainly of fibrous tissue of variable cellularity with or without cystic spaces lined by round cells. The lining cells of the cysts and the adjoining round plump cells were interpreted as true neoplastic cells of the fibroblast type. Results of light- and electron-microscopic study of human mesothelial cells and fetal mesothelial cells of rats were compared. The cytoplasmic organelles of the tumor cells were generally scanty, though rough endoplasmic reticulum, sparse mitochondria, intracellular bundles of fibrils, and numerous polysomes were seen. Some tumor cells had junctional apparatus and basement membrane and showed interdigitation of the plasma membrane. These cells lined the cystic spaces irregularly and also proliferated into the surrounding fibrous tissue, where they assumed a spindle shape and resembled fibroblasts. Ultrastructurally, the tumor cells were similar to mesothelial and stromal cells of fetal rat pleura. We speculated that the solitary (localized) mesotheliomas were probably derived from coelomic epithelium and that tumor cells remained undifferentiated or revealed minimal differentiation toward mesothelial cells.

Positron emission tomography using fluorine-18-fluorodeoxyglucose in malignant lymphoma: a comparison with proliferative activity.

To evaluate the relationship between PET using 18F-fluorodeoxyglucose (FDG) and pathological findings and to compare indices obtained by FDG-PET, FDG-PET was performed in 23 patients with untreated malignant lymphoma. Three indices obtained by FDG-PET, tumor-to-normal contrast ratio (TCR), distribution absorption ratio (DAR), k1k2/(k2 + k3), correlated with proliferative activity which was pathologically estimated both by mitotic count and by proportion of cells in all phases of the cell cycle. The relationship did not significantly change according to which of the three indices was chosen. FDG-PET, which shows the proliferative activity of tumors, is considered to be a useful method for managing tumors.

Prognostic factors for local control in nasopharyngeal cancer (NPC): analysis by multivariate proportional hazard models.

Despite the development of meticulous radiation therapy techniques, the local control of the nasopharyngeal cancer (NPC) remains unsatisfactory. We examined retrospectively the clinicopathologic factors which have impact upon local recurrence-free survival (LRFS) of the 67 patients with NPC who had been irradiated greater than or equal to 40 Gy with curative intent from 1975 through 1988. Three-year LRFS rate was 63%. T stage, histology, the presence of accompanying lymphocytic infiltration, and the properness of radiation therapy had influence upon LRFS with a statistical significance. Multivariate proportional hazard models showed that T stage and pathology retained significance. Radiation therapy properness emerged as a third factor when only the patients irradiated greater than or equal to 60 Gy were included in the analysis. From these results, three groups with low, moderate, and high risk of shortened LRFS could be separated. This risk classification could contribute to the stratification criteria of future study for the improvement of local control by new modalities.

Formation of islet amyloid fibrils in beta-secretory granules of transgenic mice expressing human islet amyloid polypeptide/amylin.

To investigate the relationship between human islet amyloid polypeptide (IAPP)/amylin expression and islet amyloid deposits in the pathogenesis of human non-insulin-dependent diabetes mellitus (NIDDM), we developed transgenic mice using a human IAPP cDNA connected to an insulin promoter. Ribonucleic acid blotting and immunohistochemistry revealed the expression of the transgene in the pancreatic beta cells. Immunogold electron microscopy showed that beta-secretory granules contained the human C-terminal flanking peptide of the IAPP precursor. Reverse-phase HPLC demonstrated human and mouse IAPP amide in the pancreas. Electron microscopy showed the accumulation of fibril-like material in a considerable number of beta-secretory granules. These results suggest that in transgenic mice, the human IAPP precursor is expressed in beta cells and becomes normally sorted into beta-secretory granules in which normal conversion to mature human IAPP takes place. The human IAPP molecules, because of their amyloidogenesis, aggregate into amyloid fibrils in secretory granules. Glucose tolerance was normal at 7 months old and islet amyloid was not observed. A longer time may be required for islet amyloid deposits and hyperglycemia to develop in mice. Our working hypothesis is that in human NIDDM, IAPP aggregates into amyloid fibrils in beta-secretory granules, and that the fibrils are released into the extracellular space and islet amyloid deposits become substantial with time.

Class II major histocompatibility antigen expression on coronary arterial endothelium in a patient with Kawasaki disease.

To investigate the class II major histocompatibility antigen expression on coronary arterial endothelium of Kawasaki disease and immunophenotypes of the infiltrating cells in the coronary vascular lesions, myocardial sections from a patient who died during the acute stage of Kawasaki disease were studied using an immunoperoxidase technique. The mononuclear cells in the lesions mainly consisted of macrophages and T cells, whereas B cells and NK/K cells were not seen. The majority of T cells reacted with Leu-3a antibodies, and only a few reacted with Leu-2a antibodies. Cells bearing the interleukin-2 receptor, indicative of activated T cells, were also found in the lesions. To determine the distribution of class II antigen, we used anti-HLA-DR antibodies. The massive expression of HLA-DR antigen on mononuclear cells was found in the lesions. In addition, the HLA-DR activation antigen was expressed on the coronary arterial endothelium at the infiltrates in which macrophages and T cells coexisted. In contrast, coronary arterial endothelium did not express HLA-DR antigens in the myocardial tissues of controls (n = 4). HLA-DR+ endothelial cells may play an important role in the development of Kawasaki vasculitis.

Benign giant cell tumor of femur with bilateral multiple pulmonary metastases.

This paper presents the unusual case of a patient with a histologically benign giant cell tumor of the femur that resulted in bilateral multiple pulmonary metastases having the appearance of benign giant cell tumor. An aggressive surgical approach was used to eradicate the pulmonary metastases; 25 nodules excised from the left lung and 33 nodules from the right lung were proved histologically to be benign giant cell tumors. Three years after bilateral thoracotomies, he remains well, without evidence of recurrent disease, and his lung function is almost normal.

Clinical episodes of granulomatous pneumonitis. Repetition during four consecutive summers.

From summer to mid-autumn for four consecutive years (1974 to 1977), a 50-year-old woman barber experienced cough and exertional dyspnea with sputum production. Rales were audible at the posterior lung bases bilaterally. Her chest x-ray film showed diffuse nodular shadows, and pulmonary function studies revealed markedly decreased DCO. Open chest lung biopsy was performed and granulomatous lesions with multinucleated giant cells without central necrosis were observed. Having experienced multiple episodes for four years, she ceased to show symptoms after demolishing and reorganizing her house. This case is thought to represent the typical summer-type hypersensitivity pneumonitis, which we have subsequently found to be the most prevalent form of hypersensitivity pneumonitis in Japan.

A case of massive true thymic hyperplasia with non-Hodgkin's lymphoma.

A case of massive true thymic hyperplasia with non-Hodgkin's lymphoma of the mediastinum is reported in a 14-year-old boy. Computed tomographic scan of the chest showed a mass of the anterior mediastinum and conspicuous swelling of the lymph nodes in the upper and lower mediastinum. They were grossly resected. The tumor of the anterior mediastinum was histologically diagnosed as true thymic hyperplasia. All of the mediastinal lymph nodes were diagnosed as non-Hodgkin's lymphoma, diffuse, mixed small and large cell type.

Pleural effusion associated with aortitis syndrome.

A patient with aortitis syndrome had a pleural effusion which subsided but reappeared with an exacerbation of aortitis symptoms while under antituberculosis treatment. The character of the fluid was that of an exudate, and the glucose concentration was normal. Clinical and laboratory features of the case suggest that the effusion was part of the aortitis syndrome per se.

Molecular genetic, cytogenetic, and immunophenotypic analyses in Castleman's disease of the plasma cell type.

Systemic Castleman's disease shows characteristic morphologic features in the lymph node and laboratory findings, but patients with this disease have variable clinical courses. The disease may constitute a spectrum of benign to malignant diseases. Thus, the clonal nature of the proliferating lymphoid cells was determined to obtain further insight into the malignant process of the disease. Two cases of systemic Castleman's disease were evaluated immunophenotypically, immunogenotypically, and cytogenetically. Both patients had a chronic relapsing clinical course. One patient had a clonal rearrangement of the immunoglobulin (Ig) lambda chain gene, but no restriction of the light chain expression was detected. This patient showed germ-line configurations of the Ig heavy-chain and T-cell receptor (TCR) beta chain genes; no detectable abnormal metaphases in the lymph node were noted. Another patient had predominance of lambda chain-positive plasma cells in the lymph node with a clonal chromosome change, but had germ-line Ig and TCR beta chain genes. The authors identified clonal changes in two cases of systemic Castleman's disease; one had a clonal immunogenotypic change and the other had a clonal cytogenetic change with an Ig light chain deviation. In both cases, however, a discordance of immunogenotypic and immunophenotypic changes was evident. Thus, the alteration may represent a type of lymphoproliferative disorder that lies between benign and malignant diseases.

Restricted V beta gene usage of tumour-infiltrating T lymphocytes in primary gastric malignant B-cell lymphoma.

Ten cases of primary gastric malignant lymphoma (PGL) were investigated by immunohistochemical and molecular genetic analysis. These cases were diagnosed histopathologically as follicular small cleaved cell type (1 case), diffuse small cleaved cell type (3 cases) and diffuse large cell type (6 cases) based on the WF (Working Formulation) classification. Seven cases classified as small cleaved or diffuse large cell type belong to low (4 cases) or high (3 cases) grade MALT lymphoma according to Isaacson's classification. All PGL belonged to B lineage cells according to immunohistochemical study and immunoglobulin rearrangements. Rearrangements of TCR beta chain genes were observed in four of the ten cases. The possibility that the TCR beta rearrangements were caused by tumour-infiltrating T-cells (TILs) was supported by the following observations: the tumours did not show T- and B-cell biphenotype, TCR beta exhibited functional VDJ rearrangement and V beta usage pattern was not a neoplastic type. Analysis of the repertoire of the TCR beta chain in TILs revealed a common usage of V beta 2 in the above four cases, and furthermore, predominant usage of a particular beta chain composed of V beta 2-D beta 2.1-J beta 2.3 was observed in one of the four cases. These results indicate that the TILs of PGL have a restricted TCR repertoire.

Parathyroid carcinoma with tracheal invasion and airway obstruction.

The following report presents an unusual case of a patient with a recurrent parathyroid carcinoma that grossly invaded the lumen of the trachea with a polypoid appearance and brought about severe airway obstruction. The three rings of the upper trachea, including the lower part of the cricoid cartilage, were resected with the tumor, and the airway was reconstructed by end-to-end anastomosis. The present case demonstrates that parathyroid carcinoma can become an intratracheal hazard, as can carcinomas of the thyroid gland, esophagus, lung, or larynx.

T cell receptor V beta repertoires of angioimmunoblastic lymphadenopathy-like T cell lymphoma.

To elucidate the pathogenesis of angioimmunoblastic lymphadenopathy-like T cell lymphoma (AILD-T) we investigated the T cell receptor V beta gene repertoires of four AILD-Ts and compared them with those of other histological types of lymphomas and three cases with reactive disorders. All lymphoma patients had rearrangement bands detected by Southern blot analysis. Only 1 of the 4 cases of AILD-T showed a single predominant usage of V beta 20 gene by PCR with 20 different V beta specific primers and the others had repertoires somewhat restricted but similar to reactive lesions. Subsequent sequencing of this PCR product revealed that only 2 of 7 clones were identical. These results suggest the monoclonal malignant cells in AILD-T are scant and that the infiltrating T cells show a reactive pattern. In the only AILD-T case with a single dominant V beta usage, the relationships of this repertoire and lymphoma cells seems to be of some consequence.

Clinicopathological evaluation of the Revised European-American Classification of Lymphoid Neoplasms (REAL) in Japan.

After the publication of a Revised European-American Classification of Lymphoid Neoplasms (REAL classification) in 1994, there have been reports from Europe and America regarding its practical utility and clinical significance. However, no studies have been published from Eastern countries including Japan. It has been well recognized that the distribution of malignant lymphoma in Japan is quite different from that seen in Western countries. In addition, some new entities have also been described in the REAL classification. Therefore, it seems important to examine its practical utility and clinical significance in Japan. Of the 579 cases reviewed, approximately 68% were B-cell non-Hodgkin's lymphoma (NHL) followed by 27% T-cell lymphomas. Hodgkin's disease (HD) comprised only 5% of all cases, making the ratio of NHL to HD 20.6. The most common type was diffuse large B-cell lymphoma which represented about 37% of all cases. Peripheral T-cell lymphomas, unspecified (PTCL), occurred in 15% whereas marginal zone B-cell lymphoma followed (14.9%). However, follicle center lymphoma (FCL) was less common (4.4%) as has been previously reported. We evaluated the clinical significance of the new REAL classification in 244 cases. International Prognostic Index (IPI) was a powerful predictor of survival (p<0.0001), and the immunophenotype was significant (p<0.05). Furthermore, here, we also attempt to establish a prognostic scheme based on the histologic type. In conclusion, the REAL classification appears to be useful and clinically significant in Japan.