Unusual clinicopathological features drew our attention to nine of 208 cases diagnosed as Hodgkin's disease. Lymph node biopsy specimens in these cases were immunostained with monoclonal antibodies against B-cell, T-cell and Reed-Sternberg cell associated antigens and epithelial membrane antigen (EMA). Reed-Sternberg-like and other atypical large cells were dispersed in a diffuse, small lymphocyte-rich background, consistent more often with the initial diagnosis of diffuse, lymphocyte predominance Hodgkin's disease. The clinical stage in these cases was unusually advanced (stages III and IV). Splenomegaly was a common feature (six of nine cases), the male to female ratio was 7:2 and the median age was 55 years (range 25-77). Response to recognized regimes for Hodgkin's disease treatment was poor in most cases, and three patients died early of their disease. Large cells were B-lymphocytes expressing EMA--an immunophenotype similar to nodular, lymphocyte predominance Hodgkin's disease. Reed-Sternberg cell and T-cell associated antigens were absent on large cells. Mature T-cells, with nuclear irregularities in some instances, predominated in the background. A more appropriate diagnostic category is, therefore, T-cell-rich B-cell lymphoma. The cases represent a 4-5% erroneous diagnosis of Hodgkin's disease and further suggest that there is a need for revision of criteria for the diagnosis of the diffuse, lymphocyte predominance variant.

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Summary The distribution of Gp 170, a multidrug resistance (MDR) associated glycoprotein, also called P-glycoprotein (P-gp), was examined by immunohistochemistry, using C219 and MRK16 monoclonal antibodies. Sixty-five tumour tissues were studied which included 40 non-lymphoid tumours, 15 chemoresistant non-Hodgkin's lymphomas and 10 Hodgkin's disease. The study was performed on both cryostat and special fixation processed and paraplast embedded (ModAMeX) sections. The latter method preserves fixationsensitive antigens such as P-gp and allows a more precise morphological identification of neoplastic and non-neoplastic cell populations in contrast to cryostat sections. Immunohistochemical expression of P-gp was expected and confirmed in many non-lymphoid tumours, but stromal macrophages and endothelial cells were also frequently stained in these cases. In non-Hodgkin's lymphomas, cells that were stained with both C219 and MRK16 monoclonal antibodies on cryostat sections were identified as macrophages and endothelial cells and not neoplastic lymphoid cells, by the ModAMeX technique. These findings suggest that the quantitative assessment of MDR RNA by Northern blotting performed on fresh homogenates overestimates the MDR content of neoplastic cells in a number of lymphoid and non-lymphoid tumours. In addition, the mechanism of chemoresistance in non-Hodgkin's lymphomas is less likely to be associated with P-gp expression.

Summary. In the search of B-cell lymphoma/leukaemia dissemination to cerebrospinal fluid (CSF), we used the highly sensitive semi-nested PCR (snPCR) for the analysis of IgH gene rearrangements. This method detects a rearranged IgH gene from a single B lymphocyte which may or may not represent the neoplastic B-cell population. We therefore performed multiple snPCR (three to five) experiments on the same CSF sample, postulating that the detection of a band of the same size and sequence in different PCR runs was highly indicative of a clonal population. 17 consecutive cases with a differential diagnosis of primary (PCNSL) (n¼10) or secondary (SCNSL) (n¼7) CNS lymphoma or leukaemia were investigated by the new strategy. The clonal nature of the B-cell population was confirmed in 3/10 of suspected PCNSL, and in six other cases the PCR study was indicative of reactive lymphocytosis. One case revealed a clonal B-cell population in the clinical context of an autoimmune disorder. Evidence of clonal B-cell population was found in 4/7 of suspected SCNSL. In one of these cases the detected band and its sequence proved identical to that of the primary nodal lymphoma. We believe that the evaluation of B-cell clonality in CSF requires multiple snPCR amplification on the same sample to compare the size of the products and, if necessary, the DNA sequences to ascertain the diagnosis of malignancy in equivocal cytologic and clinical findings.

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