Leukaemic cells from 2 patients with B-prolymphocytic leukaemia were immortalized in vitro by means of Epstein-Barr virus and phorbol-ester TPA. The resulting cell lines, named JVM-2 and JVM-3, have been growing continuously in liquid culture for more than one year. JVM-2 is characterized cytogenetically by t(11;14)(q13; q32), and JVM-3 by trisomy 12. The immunoglobulin (Ig) heavy- and light-chain genes showed the same pattern of rearrangement in both lines as in the original prolymphocytes from each case. The cells from these lines showed a spectrum of morphological and immunological features corresponding to different stages of B-cell maturation. The expression of Ig, IgM-lambda in JVM-2 and IgMD-K in JVM-3, changed from a predominantly membrane pattern in the original cells to a cytoplasmic one in the cell lines. By comparison with their original progenitors, the cells from both lines showed reduced reactivity with the monoclonal antibody (MAb) FMC7, and increased expression of the antigens recognized by the MAbs OKT10, alpha-Tac, FMC53 and Ki-I. The availability of cell lines from this rare type of lymphoid leukaemia offers a potential tool for the study of molecular events associated with the expression of Ig and other antigens by neoplastic cells.
A leukaemic phase is uncommon in large cell lymphoma, particularly at presentation of the disease, and very few cases have been reported since immunological markers became available. We have studied 24 cases presenting over a period of 15 years. 16 were B-lineage and eight T-lineage. In five patients the large cell lymphoma represented a transformation of a preceding low grade, small cell lymphoproliferative disease. Three other patients with T-lineage large cell lymphoma presented with skin infiltration (two cases) or lymphoma-associated nephrotic syndrome (one case) 3-9 months before leukaemia occurred. The other 16 patients (12 B-lineage and four T-lineage) presented with de novo large cell leukaemia/lymphoma. With the exception of skin infiltration, clinical features did not differ between T and B-lineage cases. Prognosis was generally poor although a minority of patients lived 1-2 years. Median survival was 7 months. In the B-cell lymphomas immunological markers were those expected but among the T-cell cases there were many unusual immunophenotypes with frequent failure to express markers which are usually positive in peripheral T cells. Without the availability of immunological markers diagnosis would have been difficult, with acute myeloid leukaemia being the most important differential diagnosis. T-lineage and B-lineage cases could not be distinguished on cytological features. In the majority of cases the cells were very pleomorphic with nuclear lobulation, prominent nucleoli and marked cytoplasmic basophilia being commonly observed. Marked nuclear lobulation was not confined to T-lineage cases but was seen also in seven of 12 cases of de novo B-lineage leukaemia/lymphoma; it was readily apparent in histological sections and on ultrastructural examination.
SUMMARY Ten patients with follicular lymphoma presented with a high white cell count (45-220 x 109/1) which resembled chronic lymphocytic leukaemia (CCL): all had pronounced splenomegaly and, except one, generalised lymphadenopathy. The blood lymphocytes were small with scanty cytoplasm, densely condensed nuclear chromatin, and deep clefts originating in sharp angles from the nuclear surface. CLL cells are larger, have more cytoplasm, a different pattern of chromatin condensation, and may have shallow nuclear indentations or foldings rather than clefts. The circulating follicular lymphoma cells had moderate to strong membrane immunoglobulins (SmIg), low mouse (M)-rosettes, strong reactivity with the monoclonal antibody FMC7, and occasional expression of the CD5-antigen; at least one third of cells in each case were positive with anti-cALLa (J5,CD10). Half the cases were referred as B-CLL but none had the typical B-CLL immunophenotype: weak SmIg, M-rosettes of > 50%, CD5 positive, FMC7 and J5 negative. The diagnosis of follicular lymphoma was confirmed by lymph node biopsy in seven of the 10 cases. The overall response to treatment was poor and five patients died within three years of diagnosis. This aggressive form of follicular lymphoma needs to be distinguished from B-CLL as different management is required.
We describe a method for electron microscopy that combines myeloperoxidase or acid phosphatase cytochemistry with the labelling of cell surface antigens with monoclonal antibodies and colloidal gold. This technique was tested in samples of normal mononuclear cells, leukaemic T cells with a mature or immature phenotype and an acute myeloid leukaemia. This method allows the demonstration at a single cell level of ultrastructural morphology, cytochemical reaction and the presence of a membrane antigen. It will improve further the analytic power of electron microscopy in the characterization of leukaemic cells particularly in cases of mixed leukaemias and in the study of normal haemopoietic differentiation with monoclonal antibodies.
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