Manuel E. Kaplan

Distinctive Chromosomal Abnormalities in Histologic Subtypes of Non-Hodgkin's Lymphoma

Using a new high-resolution technique for chromosomal analysis, we have successfully studied biopsy specimens of lymph nodes from 42 of 44 patients with non-Hodgkins lymphoma and have categorized them using the new international histologic formulation and immunologic markers. Abnormalities of the clonal chromosomes were detected in all 42 patients. Three recurrent chromosomal aberrations were found to correlate with certain histologic types: a translocation between chromosomes 18 and 14 in 16 of 19 patients with follicular lymphomas (small cleaved cell, mixed cell, and large cell); a translocation between chromosomes 8 and 14 in 5 of 6 patients with small noncleaved-cell (non-Burkitts) or large-cell immunoblastic lymphoma; and a trisomy 12 in 4 of 11 patients with small-cell lymphocytic lymphoma. Our findings suggest that characteristic chromosomal defects occur in certain lymphoma subtypes and that high-resolution chromosomal analysis promises to become an important tool in improving our basic understanding of lymphoid cancers.

Red-cell-membrane polypeptide aggregates in glucose-6-phosphate dehydrogenase mutants with chronic hemolytic disease. A clue to the mechanism of hemolysis.

Red-cell membranes from patients with glucose-6-phosphate dehydrogenase deficiency were studied with polyacrylamide gel electrophoresis and gel filtration chromatography in sodium dodecyl sulfate. Membranes from each of five such patients who also had chronic hemolytic disease contained polypeptide aggregates within two molecular-weight ranges (4.4 X 10(5) and greater than 50 X 10(6) daltons). The 4.4 X 10(5) dalton aggregates were not detectable in red-cell membranes of patients with the enzyme deficiency without chronic hemolysis or in membranes from normal subjects, and the greater than 50 X 10(6) dalton aggregates were not found in appreciable amounts in these cells. The aggregates were dissociated by mercaptoethanol or dithiothreitol -- indicating that they were formed by intermolecular disulfide bonds. The polypeptide aggregates contained spectrin but not globin. Red-cell deformability was decreased in aggregate-containing cells. We postulate that the polypeptide aggregates are indicators of oxidant damage to the red-cell membrane, which results in decreased deformability and chronic hemolysis.

Frequent Association of IgMλ with Crystalline Inclusions in Chronic Lymphatic Leukemic Lymphocytes

Abstract In four of 30 patients with chronic lymphocytic leukemia, studied by immunofluorescence, 1 to 31 per cent of peripheral blood lymphocytes were found to contain intracellular, rod-shaped inclusions that stained specifically for both μ and λ (IgMλ) determinants. Essentially all inclusion-containing lymphocytes exhibited monoclonal, membrane-bound IgMμ (M-IgM λ). However, only a minority of lymphocytes with M-IgM λcontained inclusions. On electron microscopy the homogeneous inclusions were found within the rough endoplasmic reticulum, and they exhibited an electrondense periodicity consistent with crystalline structure. In this series, the chance occurrence of crystals present solely in lymphocytes bearing M-IgMμ is 0.5 per cent. These studies indicate that in some patients with chronic lymphocytic leukemia, at least two morphologically distinct populations of neoplastic lymphocytes are present. (N Engl J Med 289:113–117, 1973)

Leukocytosis and large cell lung cancer. A frequent association.

In a retrospective study of 105 patients with non‐small cell lung cancer during a 5‐year period, 43 had leukocytosis. In 19 of the 43 patients, no clear cut etiology for the leukocytosis was apparent and it was attributed to the tumor itself. In these 19 patients, absolute neutrophilia was detected in 13, eosinophilia was present in three, and eleven exhibited concomitant thrombocytosis. Tumor‐associated leukocytosis occurred predominantly, and eosinophilia exclusively, in patients with large cell pulmonary neoplasms. These results suggest an unusual myeloproliferative stimulus in this type of cancer. It may result from tumor cell production of hemopoietic growth factors such as granulocyte‐macrophage colony‐stimulating activity; however, additional studies are needed to elucidate the underlying mechanism(s), and to determine whether this is a peculiar characteristic of the cells that comprise large cell undifferentiated carcinoma of the lung.

In vitro suppression of erythropoiesis by bone marrow adherent cells from some patients with fungal infection

We present evidence that alterations in marrow adherent cell (Mø) function may play a role in the suppression of erythropoiesis in some patients with fungal infection. Bone marrow (BM) cells from 12 normals and 10 patients with histoplasmosis were cultured in plasma clots before and after removal of Mø. BM from five patients (Group A) produced normal numbers of erythroid colonies (EC). In the remaining patients (Group B), smaller numbers of EC were detected (P<0.01). Removal of Mø from BM of normals and Group A patients resulted in decreased growth of EC. In contrast, Mø depletion of BM from patients in Group B resulted in greater EC formation (P< 0.01). When normal Mø were admixed with normal or patients’BM‐Mø, enhanced EC formation resulted. Whereas, at similar concentrations, Mø from group B patients caused inhibition of EC formation (P< 0.005). The erythro‐regulatory function of Mø, including the inhibitory action of patients’Mø, was mediated via a soluble agent(s) since media conditioned by Mø mimicked the action of these cells. Three patients in Group B were restudied 14 months after treatment with amphotericin B, when blood parameters had returned to normal. At this time, normal patterns of EC formation and Mø activity were observed.

Human monocyte and macrophage modulation of lymphocyte proliferation.

Abstract The effects of human monocytes and mature human macrophages on lymphocyte proliferation in response to PHA and allogeneic lymphocytes were examined. Monocytes enhanced and macrophages markedly suppressed lymphocyte proliferation to both stimuli. Monocyte enhancement of lymphocyte proliferation was, in part, due to a soluble mediator. Macrophage suppression was not due to (a) media depletion, (b) soluble lymphotoxins or inhibitors of proliferation, (c) media depletion, (d) macrophage production of prostaglandins, (e) decreased lymphocyte survival, or (f) induction of suppressor lymphocytes. These data emphasize the dichotomy of human monocyte and macrophage effects on lymphocyte proliferation and suggest, by exclusion, that macrophage suppression may require cell-cell contact.

A new method for isolation of human monocytes

Abstract A technique for obtaining purified suspensions of human blood monocytes is described. Purified monocyte monolayers (⩾95% monocytes) were prepared and then recovered into suspension by exposure to 30 mM lidocaine. Recovery of monocytes from adherent monolayers was quantitative and no significant loss of viability occurred. Once lidocaine was removed from cell suspensions, function of purified monocytes was equivalent to monocytes in the original unpurified cell preparations. This technique should be of value in future studies of human monocytes.

Terminal transferase levels in chronic myelogenous leukemia in blast crisis and in remission

Abstract Terminal deoxynucleotidyl transferase (TdT) has been cited as a possible biological marker for acute lymphoblastic leukemia (ALL) and the blast phase of chronic myeloid leukemia (CML). In the present study high levels of TdT, comparable to levels present in normal thymus, were observed in circulating leukocytes of a CML patient in blast crisis. Sequential studies revealed disappearance of TdT from circulating leukocytes after remission was induced with vincristine and prednisone therapy. Blood TdT returned to moderately elevated levels in the absence of detectable circulating blasts, 5 months before the reappearance of blast crisis. During blast crisis the malignant cells were devoid of myeloid features on light and electron microscopy and were peroxidase negative and periodic acid-Schiff (PAS) positive. These studies demonstrate that elevated TdT levels may occur with or without detectable morphologic evidence of circulating blast cells. The presence of complement receptors on agranular blasts during the blast phase of chronic myeloid leukemia is a new finding.

Two Mechanisms of Erythrocyte Destruction in Penicillin-Induced Hemolytic Anemia

Abstract Antiglobulin testing revealed IgG, C3, C4 and C5 on the erythrocytes in a case of penicillin-induced, immune hemolytic anemia. When no penicillin was in the patients serum, the survival times of autologous and of normal erythrocytes, measured concomitantly, were both shortened (half-time of 21 days). Antibody specific for penicillin was eluted from patient erythrocytes. On addition of eluate to a mixture of normal B cells coated with penicillin and O cells in the presence of complement, C3, but not IgG, was bound to the O cells. Addition of eluate and complement to a mixture of O erythrocytes treated with 2-amino-ethylisothiouronium bromide and penicillin-coated B cells resulted in hemolysis of the O cells. Two mechanisms of erythrocyte destruction are possible: the IgG, complement-binding, antipenicillin antibody causes destruction of patient erythrocytes coated in vivo with penicillin; and, in the absence of detectable serum penicillin, normal erythrocytes may be destroyed because they bind ac...

Non-Hodgkin’s Lymphoma after Treatment for Hodgkin’s Disease

A patient is described who developed a diffuse histiocytic lymphoma (DHL) 9 years after radiation therapy for Hodgkins disease. This occurrence is of particular interest because the treatment for Hodgkins disease included no chemotherapy and the second tumor appeared to originate remote from the irradiated site. Thus, the role of Hodgkins disease treatment in the etiology of this patients second lymphoma appears doubtful. The development of DHL in this patient could represent an unrelated event or conceivably an event facilitated by Hodgkins disease itself.