Yoshiaki Sumiyoshi

Analysis of Epstein-Barr viral genomes in lymphoid malignancy using southern blotting, polymerase chain reaction and in situ hybridization

Summary109 malignant lymphomas were surveyed by Southern blot analysis and polymerase chain reaction (PCR) for Epstein-Barr virus (EBV) DNA and compared with 16 examples of non-neoplastic lymphadenopathy and 4 normal thymuses. In specimens positive by the method of Southern and PCR, in situ hybridization studies were performed on formalin-fixed, paraffin-embedded sections. By Southern blot analysis, two of seven Hodgkin’s disease samples (29%) (one of mixed cellularity and the other of lymphocyte predominance type), three of 56 B-cell lymphomas (5.6%) and five of 46 Tcell lymphomas (11 %) demonstrated EBV DNA. However, the 16 examples of lymphadenitis and the 4 normal thymuses showed no EBV DNA. With PCR, EBV DNA was identified in one B-cell lymphoma, nine T-cell lymphomas, ten lymphadenitis specimens and two of the normal thymus, in addition to the positive specimens determined by the Southern blotting method. These results indicate that the presence of EBV DNA is not related to lymphoid malignancy, but enhancement of the DNA is demonstrated in some neoplastic conditions. By in situ hybridization, EBV genomes were not detected in all PCR-positive cases, but only in those positive by Southern blot analysis.

A case of histiocytic necrotizing lymphadenitis with bone marrow and skin involvement

We report a case of histiocytic necrotizing lymphadenitis (HNL) with bone marrow extension in a 29-year-old male in which many large mononuclear cells infiltrated the bone marrow and mimicked malignant lymphoma. A lymph node biopsy confirmed the diagnosis of HNL. Immunohistologically, the infiltrating cells in the bone marrow were positive for lysozyme, LeuM1, Kp-1 and T-cell markers. The cells did not show haemophagocytosis. A skin biopsy from an accompanying facial skin rash revealed a proliferation of large cells similar to those observed in affected foci of the lymph node in subcutaneous tissue. The infiltrating cells were mainly lysozyme and Kp-1-positive histiocytes, some with phagocytosis of nuclear debris but none characteristic of haemophagocytosis. Transformed T-cells were also infiltrating.

Alpha-interferon in Kikuchi's disease

SummaryIn Japan, histiocytic necrotizing lymphadenitis (Kikuchi’s disease) is a relatively common reactive lesion affecting lymph nodes, but the histogenesis and pathogenesis of the disease have not been clarified. Alpha-interferon has a role in the body’s defense against, viral infections. Using a polyclonal antibody to human alpha-interferon, we found numerous cells, mainly histiocytes, containing alpha-interferon in affected foci in the lymph nodes from 24 patients with Kikuchi’s disease. Tubuloreticular structures, thought by some authors to be associated with the production of interferon, were detected by electron microscopy in histiocytes, activated lymphocytes and vascular endothelial cells in the affected foci. These results suggested that the formation of tubuloreticular structures is a secondary phenomenon following stimulation by alpha-interferon. Further, the activity of 2′–5′ oligoadenylate synthetase, which is induced by alpha-interferon and enhanced during the early or active stage of viral infection, showed increased levels of activity in the active stage of Kikuchi’s disease and decreased to normal levels in the convalescent stage 2 weeks later. These results suggested the possibility of a viral etiology for Kikuchi’s disease.

Proliferating cells in histiocytic necrotizing lymphadenitis.

SummaryThe phenotypes of proliferating cells in histiocytic necrotizing lymphadenitis (HNL) were examined. The affected areas consisted mainly of CD 8-positive (suppressor/cytotoxic T-cells) and CD 4-positive (helper/inducer T-cells) in association with some CD 15-positive cells (monocytes). A marker of proliferating cells (Ki-67) and monoclonal antibodies for determining the phenotypes of cells (CD 4, CD 8, CD 15) in the affected areas were applied using a double-staining method. Ki-67-positive proliferating cells were mainly CD 8-positive. A few CD 4-positive cells and rare CD 15-positive cells were also Ki-67-positive. The percentage of CD 8-positive cells increased gradually over time and the ratio of CD 8-positive to proliferating cells did not decrease throughout the observation period of 6 weeks. These results suggest that the proliferation of CD 8-positive T-cells together with the accumulation of CD 4- and CD 15-positive cells is the main phenomenon occurring in HNL.

Analysis of herpesvirus genomes in Kikuchi's disease

We examined the cervical lymph nodes of 30 patients with Kikuchis disease and 15 patients with nonspecific lymphadenitis, using Southern blot analysis and polymerase chain reaction (PCR) to identify human herpesviruses such as Epstein-Barr virus (EBV), cytomegalovirus, herpes simplex virus, and varicella-zoster virus. By Southern blot analysis, no virus DNA was recognized, but 16 of the 30 nodes from patients with Kikuchis disease and 8 of the 15 nodes from patients with non-specific lymphadenitis showed amplified EBV DNA by PCR.

Immunohistological study of skin involvement in Kikuchi’s disease

SummaryFive patients with histiocytic necrotizing lymphadenitis (Kikuchi’s disease) with erythematus or popular skin lesions were studied. All patients healed naturally within a few months like the patients with no skin involvement. Histological findings for the skin lesions mimicked cutaneous malignant lymphoma. The infiltrated mononuclear cells usually demonstrated positive reactions for Ki-M1p (20–63%), lysozymes (13–54%), MT-1 (18–64%), UCHL-1 (22–39%) and LN2 (17–36%). OPD-4 and L26 positive cells were few in number. These results suggest that the infiltrated cells in a Kikuchi’s disease skin lesion are composed of the same components as the affected lesion in the lymph node.

Expression of CD44, vascular endothelial growth factor, and proliferating cell nuclear antigen in severe venous invasional colorectal cancer and its relationship to liver metastasis.

Abstract: The first step in liver metastasis is venous invasion by cancer cells from the primary tumor. However, even among cases where the histology shows extensive venous invasion by the primary tumor, we sometimes find cases without synchronous liver metastases. As a result, there is a strong possibility that, besides the established causes of colorectal cancer and that of cancer cells invading the veins, some other important causes for liver metastasis must exist. We investigated the expression rates of CD44, proliferating cell nuclear antigen (PCNA), and vascular endothelial growth factor (VEGF) in 28 primary colorectal tumors using immunohistological techniques, and examined an association with liver metastasis. Cases that are strongly positive for CD44 or PCNA have a higher rate of synchronous liver metastases than cases with either no expression or a low expression. We could find no correlation between the VEGF expression and synchronous liver metastasis. In cases with severe venous invasion, VEGF is not correlated with liver metastasis whereas CD44 and PCNA are correlated with liver metastasis. In cases where severe venous invasion is histologically observed, an immunohistochemical analysis for CD44 and PCNA should be done to assess the likelihood of liver metastases.

Genetic changes in atypical hyperplasia and lymphoma with angioimmunoblastic lymphadenopathy and dysproteinaemia in the same patients

The transition between atypical hyperplasia and lymphoma with angioimmunoblastic lymphadenopathy and dysproteinaemia (AILD) was studied in serial lymp node biopsy specimens from five patients using DNA analysis with Southern blot analysis, polymerase chain reaction, chromosomal analysis, and immunophenotyping. The chromosomal analysis showed additional abnormalities as the disease progressed to those present initially, and immunological staining showed a corresponding increase in the numbers of CD4- and Ki67-positive cells. In the first biopsy from each patient a diagnosis of atypical hyperplasia with AILD was made and lymphoma excluding by the finding of only a few atypical lymphoid cells and the preservation of follicles with germinal centres. DNA analysis of lymph nodes at this stage showed either germ lines or oligoclonal rearrangements of the T-cell receptor (TCR) and immunoglobulin heavy chain genes. In the final biopsy, when a diagnosis of lymphoma with AILD was made, either a monoclonal rearrangement of the TCR was observed or one of the rearranged bands had increased in density. These results suggest selective proliferation of a clone of abnormal cells may account for the progression of atypical hyperplasia to lymphoma with AILD.

Analysis of human herpes virus-6 genomes in lymphoid malignancy in Japan.

Ninety cases of malignant lymphoma and 56 cases of reactive lymphadenopathy were studied using Southern blot analysis and the polymerase chain reaction to identify human herpes virus-6 (HHV-6) DNA. This was detected in cases of lymphoid malignancy at a rate which ranged from 50.0% to 68.8%. There were no differences in rates for different types of lymphoid malignancies. Herpes virus-6 DNA was detected by PCR in lymphoid malignancies less frequently than in reactive lymphadenopathies. It was not detected in lymphoid malignancies using Southern blotting. These results suggest that HHV-6 DNA was not related to lymphoid malignancy and was only a latent infection of non-neoplastic cells in tumour tissue.

A solitary peutz-jeghers type hamartomatous polyp in the duodenum — A case report—

A 56-yr-old woman with a solitary hamartomatous polyp confined to within the third portion of the duodenum, underwent a successful endoscopic polypectomy. The histological findings were identical to Peutz-Jeghers polyp but all the other clinical features of Peutz-Jeghers syndrome were absent.