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We applied a newly developed ultrastructural cytochemical technique utilizing 2-naphthyl thiol acetate (NTA) as a substrate in 3 cases of true histiocytic-monocytic malignancy and 3 controls. The enzyme NTA esterase was identified in both neoplastic and benign histiocytes-monocytes in the form of electron-dense material within the cytoplasm. The NTA esterase varied in size and shape and showed no clear relationship with cytoplasmic organelles. The distribution and pattern of staining were very similar to those of the light-microscopically demonstrated alpha-naphthyl acetate esterase reaction. The main advantages of this method are: 1) histiocytes-monocytes can be identified in the absence of lysosomes or phagocytosis; 2) unequivocal, simultaneous documentation of both histiocytic esterase and subcellular structure is possible; and 3) both blood and solid tissue specimens can be utilized. Our findings demonstrate the usefulness and applicability of ultrastructurally demonstrated NTA esterase in the study of histiocytic-monocytic malignancies.

Two distinct differentiation antigens of human myelomonocytic cells are defined using murine monoclonal antibodies. The antigens recognized by antibodies 20.2 and 20.3 are expressed by all cells of the monocyte lineage in both peripheral blood and bone marrow. Cell-sorting experiments demonstrated that histiocytes and immature bone marrow cells with detectable alpha-naphthyl butyrate esterase activity also express both antigens. Within cells of other lineages, the antigens had distinct patterns of expression. Immature myeloid cells were 20.2 negative, but 20.3 positive; whereas mature myeloid cells were 20.2 positive, but 20.3 negative. Nucleated erythroid cells and platelets expressed only the 20.3 antigen. These results indicate that myeloid and monocytic cells share common differentiation antigens with cells of the erythroid and megakaryocytic lineages. The 20.2 and 20.3 antibodies reacted with the leukemic cells from some patients with acute nonlymphocytic leukemia (FAB, M1-M5) and with some cell lines derived from patients with nonlymphocytic leukemia, but not with blast cells from patients with lymphoid leukemia or with lymphoid leukemic cell lines. These antibodies may prove useful in studying the differentiation of bone marrow stem cells, in defining the cellular origins and classification of leukemias, and in the identification of distinct prognostic subgroups of acute nonlymphocytic leukemia.

Tocainide is an effective oral antiarrhythmic agent. We report a 77-year-old man who developed agranulocytosis and anemia while receiving tocainide therapy. These hematologic abnormalities were detected on routine evaluation six weeks after beginning tocainide therapy. The absolute granulocyte count decreased to 50/mm3 (0.05 X 10(9)/L). The anemia was mild; hemoglobin count, 10.9 g/dL (109 g/L). These abnormalities were associated with local and stromal adipocytic bone marrow damage, and decreased production of red blood cells and granulocytes. The platelet count was not affected. The patient had no evidence of infection. Hematologic values were restored to normal two weeks after discontinuation of tocainide therapy, indicating that bone marrow toxicity of tocainide is reversible.

In a comparative immunologic and cytochemical analysis, Reed-Sternberg (RS) cells from seven patients with Hodgkin's disease were found to more closely resemble interdigitating reticulum cells (IDC) than normal or malignant histiocytic reticulum cells (macrophages); malignant macrophages were established as a cell line from a patient with malignant histiocytosis. RS cells and IDC both showed strong expression of human Ia-like antigens, a close physical association with helper/inducer T cells, and a lack of at least two common macrophage antigens. Supportive evidence of the identity of RS cells and IDC was their lack of lymphoid and myeloid characteristics smaller content of nonspecific esterase and acid phosphatase than that in histiocytes, similar cytology, and location in thymic-dependent regions of lymphoid tissues. Malignant transformation of the IDC to an RS cell could result in diminished antigen-presenting capacity and contribute to the defect in T-cell-mediated immunity that commonly occurs in Hodgkin's disease.

Clinical and pathological studies of cutaneous T-cell lymphomas (CTCL) reveal variations in tumor cell morphology and surface membrane phenotype that are of diagnostic and prognostic importance. Our study investigates blastic transformation and surface antigen change on CTCL cells in vitro under the influence of tumor-promoting phorbol ester (TPA) and phytohemagglutinin. Both agents transformed tumor cells with cerebriform nuclei into blast cells within 5 days; however, Sézary cells were somewhat resistant to transformation with phytohemagglutinin. Multinucleated cells with prominent nucleoli resembled Reed-Sternberg cells of Hodgkin's disease. These morphological changes simulated the appearance of the aggressive tumor stage of mycosis fungoides. During blastic transformation, the erythrocyte rosette receptor was induced by TPA on sheep erythrocyte-rosette-negative Sézary cells from one patient. During the first 24 hr in vitro, Sézary and MF cells stimulated by TPA lost Leu 3a (T4) antigen while maintaining original high levels of Leu 1 antigen. In contrast, leukemia cells from patients with adult T-cell leukemia (ATL) were resistant to modulation of Leu 3a antigen by TPA; 3A1 antigen on CTCL and ATL cells was unaffected by TPA. Blastic transformation of CTCL cells was observed with both TPA and phytohemagglutinin, but helper T-cell antigen Leu 3a (T4) and erythrocyte rosette receptor changes occurred only with TPA. Thus, blastic transformation and surface differentiation were not directly related. These results provide a possible model for the study of blastic transformation and surface antigen/receptor variation in CTCL. They also may provide an independent test for the distinction of CTCL and ATL in vitro. Finally, they illustrate the relative resistance of ATL to surface antigen modulation as previously shown for Tac antigen modulation by anti-Tac antibody.

Adult T-cell leukemia/lymphoma (ATLL) is a recently described distinct clinicopathologic entity characterized by a leukemic or lymphomatous proliferation of hyperlobulated peripheral T-cells, which is usually widespread at presentation and is associated with infection by a type C retrovirus. ATLL rarely is described outside of endemic regions, which include southwestern Japan, the Caribbean region, and the southeastern United States. The authors report the clinical, pathologic, and immunologic features of two cases of nonendemic ATLL that occurred in patients from the midwest United States. One patient was a 16-year-old white girl from rural Iowa, and the other was a 46-year-old white man from rural Minnesota. The features of 13 other probable nonendemic ATLL cases from the United States were compiled and reviewed. In the United States, nonendemic ATLL occurred in widespread geographic locations, affected mostly white people, and was characterized by an aggressive course with generalized adenopathy, blood and bone marrow involvement, and hepatosplenomegaly at presentation. Skin involvement was present in one-fourth of the patients. Hypercalcemia was rare. Although antibodies to type C retrovirus were detected in three of the five patients tested, the available data is not sufficient to establish a conclusive association between nonendemic ATLL and type C retrovirus infection.

Four young women presented with diarrhea, malabsorption, and intestinal pseudoobstruction. Intestinal biopsy specimens (both peroral and full-thickness) showed flat small intestinal mucosa, sparsity of crypts, and a widespread lymphoid infiltrate in the lamina propria, muscularis propria, and myenteric plexus. There was no neuron or nerve fiber loss or damage in the plexus; muscle cell absence in the vicinity of lymphoid cell infiltration in the muscularis propria probably accounted for the pathogenesis of pseudoobstruction. Immunochemical stains showed that the infiltrate was polyclonal, and none of the patients has developed lymphoma on clinical follow-up of 4-16 yr. Transient improvement in symptoms occurred after antibiotic therapy in 3 patients, and 1 patient had improvement after treatment with cyclophosphamide and prednisone; however, symptoms of pseudoobstruction persist in all. These cases illustrate yet another cause of intestinal pseudoobstruction which is histologically distinct from visceral myopathies and neuropathies. The pathogenesis of this illness may be related to that of diffuse immunoproliferative diseases seen in Third World countries.

Neoplastic cells from 13 cases of hairy cell leukemia were investigated for immunoglobulin production and lysozyme activity by an electron-immunoperoxidase technique. In 10 cases cytoplasmic immunoglobulins were found, but lysozyme activity was absent in all cases. Immunoglobulins were detected in the perinuclear space and endoplasmic reticulum and at the surface of hairy cells. Of the cases in which immunoglobulins were detected in hairy cells, nine were positive with IgM antiserum and one with IgG antiserum. The immunoglobulins were monoclonal in all cases; six were positive with lambda antiserum and three with kappa antiserum. The class and type of surface immunoglobulins were identical to those of cytoplasmic immunoglobulins in the hairy cells. These results support the conclusion that hairy cells are commonly derived from immunoglobulin-producing B cells at an earlier stage of differentiation than plasma cells.

Ricinus communis agglutinin (RCA) staining of malignant histiocytosis (two cases) and tissue infiltrates of monoblastic leukemia revealed two distinct macrophage-histiocyte types: one with cytoplasmic staining (RCA+), the other without (RCA-). RCA+ cells corresponded to benign-appearing histiocytes without nuclear atypia, whereas RCA-cells were cytologically malignant. In one case of malignant histiocytosis, many RCA-tumor cells were surrounded by a thin cytoplasmic extension of RCA+ cells, which suggested an interaction between the tumor cells and stromal macrophage-histiocytes. These observations support the view that most benign-appearing histiocytes in malignant histiocytosis are reactive in nature and suggest that RCA staining can be useful in distinguishing between benign and malignant disorders of macrophage-histiocytes.

This paper describes a newly recognized clinicopathologic syndrome of regressing cutaneous nodules, peripheral lymphadenopathy and fever occurring mainly in children and adolescents. Skin lesions show pseudoepitheliomatous hyperplasia, absence of epidermotropism and pleomorphic large lymphoma cells infiltrating the dermis and subcutaneous fat. Lymph nodes have a distinctive appearance with lymphoma cells in sinuses and paracortex. Despite their usual histiocyte-like appearance, the lymphoma cells in most cases have an aberrant T-cell phenotype, expressing several activation antigens including Hodgkin's disease associated antigen Ki-1 (CD30). Clonal rearrangements of the beta chain genes for the T-cell antigen receptor (TCR) were found and revealed a T-cell origin in one case lacking T-cell surface antigens. Identical rearrangements of the TCR genes indicated a monoclonal origin for separate lesions on the arm and leg of another patient. Treatment results in the first group of patients indicate that radiation treatment of an affected area is associated with frequent relapse at other sites, but multiagent chemotherapy (COMP or D-COMP) produces durable remissions in nearly all patients. Bone marrow transplantation was effective salvage therapy for treatment failures. Further studies are underway to investigate the mechanism(s) of tumor regression, the epidemiology, and etiology of Ki-1+ lymphomas which appear to be relatively more frequent in Oriental and Black patients, and in which the first lesion may resemble an insect bite.

Three patients had leukocytosis of large granular lymphocytes and chronic neutropenia. Clonal chromosomal abnormalities (trisomy 8 and trisomy 14) and lymphocytic infiltration of splenic red pulp, hepatic sinusoids, and bone marrow indicated the neoplastic nature of the large granular lymphocytes. Demonstration of a T3+, T8+, HNK-1 + phenotype and low natural killer cell activity that was augmented by interferon treatment showed the leukemic cells to be immature natural killer cells. Multiple autoantibodies were present and included rheumatoid factor and antinuclear, antineutrophil, antiplatelet, and antierythrocyte antibodies, suggesting a defect of B-cell immunoregulation. In addition, in-vitro studies showed impaired suppression of immunoglobulin biosynthesis by abnormal cells from one patient. Antineutrophil antibodies and absence of direct cell-mediated inhibition of granulocyte-macrophage colony formation supported a humoral immune mechanism for the neutropenia. In these patients the syndrome of splenomegaly, multiple autoantibodies with neutropenia, and lymphocytosis of large granular lymphocytes is due to a neoplastic proliferation of immature natural killer cells.

To investigate the prognostic value of histologic and clinical factors in lymphomas of the gastrointestinal (GI) tract, the clinicopathologic findings in 56 Japanese patients with GI lymphomas were reviewed. They included 37 patients with gastric and 19 with intestinal lymphomas. The male to female ratio was 2.7:1 in gastric and 11:1 in small intestinal lymphomas. Histologically, all but one of intestinal lymphomas were high-grade lymphomas. Gastric lymphomas comprised 47% of low-grade and 53% of high-grade tumors. Significant factors for favorable prognosis identified by Cox's multivariate analysis were female sex, the presence of reactive lymphoid hyperplasia (RLH) in the primary site, early stage of disease, gastric lymphomas, and low-grade histologic type. The important role of chronic lymphocytic infiltration for development of gastric lymphoma was suggested by the high incidence of RLH and the intermediate lymphocytic type of lymphoma.

Comparison of LyP and HD suggests that these are closely related lymphoproliferative disorders that may occur in the same patient, share similar histologic features, and have the same immunologic phenotype. Recent studies of the molecular biology of both disorders confirm a similar distribution of T-cell receptor gene rearrangement patterns. The study of LyP, although a rare and benign disorder, has improved our understanding of HD, which appears, in some cases, to be a proliferation of malignant activated T lymphocytes. Further analysis of LyP may lead to a better understanding of the spontaneous regression of this disorder and result in new methods for treatment of HD.

Stabilization of cell surface antigens and preservation of tissue morphologic characteristics are important for diagnostic immunologic studies. Current reports continue to regard unfixed frozen sections as the material of choice for immunoperoxidase studies of lymphoproliferative diseases. In this study, periodate-lysine-paraformaldehyde (PLP) is shown to be a valuable fixative for the improved detection of surface antigens in lymphoid tissue. In cases of non-Hodgkin's lymphoma and Hodgkin's disease, more frequent detection of diagnostic markers and ease of interpretation was demonstrated by use of PLP-fixed frozen tissue as compared with unfixed frozen tissue. Immunoglobulin staining was more easily interpreted in 30% of B-cell non-Hodgkin's lymphoma. In Hodgkin's disease, Ki-1 antigen, a diagnostic marker of Reed-Sternberg cells, was found in PLP-fixed tissue from two cases in which this antigen was not detected in corresponding unfixed frozen tissue. The authors have demonstrated that PLP-fixed tissue can be sent to a central reference laboratory at ambient or room temperature, avoiding the expense and inconvenience of transporting specimens on dry ice. The authors conclude that PLP fixation is the preferred method for immunopathologic study of human lymphomas.

To understand the molecular basis for aberrant expression of T cell antigens in T cell malignancies, the authors studied the immunophenotypes, rearrangements of T cell receptor (TCR)-beta chain genes, and transcription of TCR-alpha and -beta, CD3(T3), and CD2(T11) genes in 5 postthymic activated T cell lines, 2 lymphoblastic T cell lines, and 2 histiocytic cell lines. Rearrangements of TCR-beta genes and functional 1.6 kb transcripts of TCR-alpha gene were demonstrated in each of the 7 T cell lines, but not in the 2 histiocytic cell lines. There often was incomplete transcription of TCR-beta chain and CD3-delta genes in postthymic T cell malignancies. At least 3 patterns could be defined: Pattern A, presence of TCR-beta and CD3-delta transcripts; Pattern B, presence of CD3-delta but lack of TCR-beta transcripts; and Pattern C, lack of both TCR-beta and CD3-delta transcripts. Expression of TCR-beta gene could be induced by phorbol ester in 3 cell lines from postthymic T cell malignancies, indicating that there was functional rearrangement of the TCR-beta gene. CD2 transcripts were detected in 1 cell line from T lymphoblastic and 1 postthymic activated T cell malignancy. No transcripts of TCR-beta, CD2, and CD3 genes were detected in the 2 malignant histiocytic cell lines. These studies demonstrate a molecular mechanism for the aberrant expression of T cell antigens in malignancies derived from postthymic activated T cells and indicate that the detection of TCR-alpha transcripts may be useful for recognizing anaplastic T cell malignancies and distinguishing them from those of true histiocytic origin.

The pathogenesis of pure red cell aplasia (PRCA) was studied in a patient who had no evidence of malignancy. In marrow culture, no erythroid colonies (from late erythroid progenitors [CFU-E]) but normal numbers of well-haemoglobinized erythroid bursts (from early erythroid progenitors [BFU-E]) were found, indicating that BFU-E existed in the patient but that their subsequent in vivo differentiation was inhibited. Autologous coculture studies suggested that inhibition was mediated by the patient's ER + lymphocytes. After remission was induced with cyclophosphamide, autologous ER + cells no longer suppressed in vitro erythropoiesis. However, cryopreserved ER + cells, obtained with anaemia, suppressed BFU-E growth from remission marrow. An expanded population of large granular lymphocytes (LGL) with ER +, Fc gamma +, T3+, T8+, HNK-1+, Ia-, M1 -- phenotype and no functional natural killer (NK) cell activity was noted during PRCA that reverted to normal with remission. For this patient, both in vivo and in vitro evidence demonstrates a cellular inhibition of erythropoiesis at the level of differentiation between BFU-E and CFU-E.

The prognosis of acute lymphoblastic leukemia is affected adversely by T-cell markers on the malignant lymphoblasts. Direct involvement of the thymus has been assumed because of frequent mediastinal enlargement in these patients and the important role of the thymus in certain mouse leukemias. We report here an adult with T-cell leukemia and mediastinal enlargement who had physiologic involution of the thymus. The possible role of diphenylhydantoin in the pathogenesis of this disease is reviewed. The possible usefulness of Nu/Nu mouse xenografting for the study of this lethal disorder is discussed.

Five patients with polyarthritis and neutropenia had numerous circulating large granular lymphocytes with a phenotype attributed to immature natural killer cells. All five had splenomegaly and recurrent infections. Arthritis was most prominent at the wrists and hands, and all patients were considered to have atypical cases of Felty's syndrome. Antinuclear antibodies, rheumatoid factor, antineutrophil antibodies, and immune complexes were detected in most patients. Bone marrow biopsies revealed a maturation arrest at the myelocyte stage and lymphoid infiltrates. Large lymphocytes with azurophilic cytoplasmic granules were found on peripheral blood smears and showed a characteristic reactivity pattern with monoclonal antibodies suggesting a natural killer cell lineage. Peripheral blood mononuclear cells showed less than normal natural killer activity against K562 target cells. Increased numbers of large granular lymphocytes with a phenotype of immature natural killer cells may be important in the pathogenesis of neutropenia, humoral immune disturbances, and synovitis in a subset of patients with Felty's syndrome.

The authors conducted a histopathologic study on tissues from 11 patients with the recently described syndrome of large granular lymphocyte (LGL) leukemia. Distinctive pathologic findings were found most often in the bone marrow, liver, and spleen. The bone marrow biopsies contained nonparatrabecular lymphoid infiltrates that were nodular or diffuse and interstitial. Plasmacytosis was found and in two cases there was myeloid maturation arrest. The liver biopsies contained sinusoidal and portal infiltrates and the spleen had red pulp cord and sinus infiltrates, plasmacytosis, and follicular hyperplasia. Lymph node involvement was nondiagnostic, consistent with the usual absence of lymphadenopathy. The morphologic findings were sometimes indistinguishable from other reactive or low-grade lymphoproliferative disorders, especially chronic lymphocytic leukemia/well-differentiated lymphocytic lymphoma (CLL/WDLL) and hairy cell leukemia. These results suggest the need to correlate peripheral blood cell counts and morphology as well as immunophenotypic studies with tissue histology to distinguish LGL leukemia from other disorders. Establishing a correct diagnosis of LGL leukemia may help clarify the etiology of unexplained peripheral blood cytopenias, arthritis, and other autoimmune manifestations in individual patients.

A cDNA of the C beta 2 gene of the T-cell receptor was used as a probe to investigate the clonal composition of T cells in skin lesions of 5 patients with lymphomatoid papulosis (LyP), a chronic recurrent eruption characterized by morphologically abnormal activated T cells in the cutaneous infiltrate. Clonal T-cell populations, as evidenced by rearranged DNA bands, were demonstrated in the skin lesions of four patients, one of whom has shown clinical progression toward lymphoma. Three of these patients had lesions of type A histology, a type previously shown to be associated with aneuploidy. The remaining patient with clonal lesions appeared to have the same gene rearrangement pattern in DNA obtained from separate lesions taken 11 months apart, providing evidence that the T cells in both sites were derived from the same clone. This patient had lesions of type B histology, which is not associated with aneuploidy. Absence of a rearranged band and deletion or near absence of the 10.8 kb band in Eco RI digests was interpreted as evidence of polyclonal T-cell hyperplasia, accounting for the skin infiltrate of a fifth patient who had a prolonged clinical course without progression to lymphoma. This patient had lesions of type A histology with frequent Ki-1-positive Reed-Sternberg-like cells. Our results show that gene rearrangement analysis provides information that is independent of histology in LyP and may in part explain the variable progression of LyP to lymphoma in 10-20% of patients.

Expression of T-cell antigens by Reed-Sternberg (RS) cells has not been detected in most studies of Hodgkin's disease (HD). The authors employed an improved method of fixation (paraformaldehyde-lysine-periodate), which sharply defined cell borders and revealed T-cell antigens on RS cells in 8 of 30 (27%) cases of HD. Antigen-specific staining was confirmed by immunoelectron microscopy. RS cells expressed T11 (8/8 cases), Leu-3 or T4 (4/8 cases), Leu-4 or T3 (3/8 cases), but not other T-cell specific antigens (Leu-1, T8, T6, 3A1). RS cells were negative for leukocyte common antigen (LCA/T200), in contrast to positive LCA/T200 staining of RS-like cells in T-cell lymphomas. RS cells in all HD cases were positive for Ki-1 and/or Leu-M1 antigens. The percentage of RS cells expressing T-cell antigens was less than 20% (2 cases), 20-50% (3 cases), or greater than 50% (3 cases). This percentage and the specific T-cell antigens expressed varied in tissues from different sites in each of 2 cases. Expression of T-cell antigens by RS cells was found in nodular sclerosis (6 of 20 cases) and mixed cellularity (2 of 5 cases) but not in lymphocyte predominance (2 cases), lymphocyte depletion (1 case), or unclassified types (2 cases). Two cases of nodular sclerosis contained areas of necrosis surrounded by sheets of lacunar cells (syncytial variant of NSHD). Two other cases were associated with cutaneous lymphoma. One of these cases was mixed cellularity HD, which appeared to be confined to the skin. In a second case, tumor cells of similar phenotype (T4+, Ki-1+) were found in skin and lymph nodes of a patient with coexistent mycosis fungoides and HD. These results are consistent with an origin of RS cells from T cells in some cases of nodular sclerosing and mixed cellularity HD. They also suggest that the same cell type, an activated helper T-cell, is involved in the pathogenesis of both skin lesions and lymphadenopathy of some patients with coexistent mycosis fungoides and HD.

Extracorporeal photopheresis (ExP) was administered every other week in an outpatient setting to four patients with chronic refractory psoriasis vulgaris without arthropathy. The duration of treatment ranged from six to 13 months. Two patients received methotrexate concomitantly during the initial phase of the study. All patients demonstrated a decrease in erythema, induration, and scaling of lesional skin, accompanied by incomplete clearing of lesions such that the percentage of involvement (SI) ranged between 40 to 80 percent of baseline scores. Exacerbations of psoriasis occurred with minor provocations, and two patients who were predisposed to developing epithelial skin neoplasms as a consequence of prior treatments continued to develop tumors during the study interval. Prolonged ExP treatment was otherwise well tolerated, without evidence of toxicity on routine laboratory safety tests or changes in lymphocyte counts. All patients, however, exhibited decreased intradermal skin responses to recall antigens and a decreased capacity of peripheral lymphocytes to produce interleukin 2 (IL-2) in response to polyclonal stimuli in vitro. These observations suggest that the observed anti-inflammatory effect of alternate-week ExP on psoriasis is mediated in part to a direct inhibition of lymphokine production or release by psoralen-ultraviolet-exposed lymphocytes.

Using electron microscopy and cytochemical techniques, the authors determined the distribution of acid phosphatase (AcPase) within the organelles of lymphocytes from blood rosetted with either neuraminidase-treated sheep erythrocytes (En) or sheep erythrocytes coated with antibody and complement (EACs). Subsequently, the various reactive organelles of the rosetted lymphocytes were counted, affording a comparison of T and B cells. It was found that AcPase was present in approximately 80% of T cells and 45% of B cells and was most frequently observed in secondary lysosomes of varying size and content. Although more T cells than B cells were reactive for AcPase, the extent of reaction in some B cells clearly precludes the use of AcPase for differentiating the two cell lines. It should be recognized that while the En rosetting procedure detects T cells in a nonselective manner, the EAC rosette is a marker of a major subpopulation of B lymphocytes, ie, those bearing complement receptors. We believe that the distribution of lysosomal enzymes in B and T lymphocytes probably reflects the functional state of individual cells rather than being a reliable indicator of cell lineage. A surprising finding (which could be established only by a fine-structural study) was the fact that 20% of circulating "resting" T cells contained reaction product for AcPase within endoplasmic reticulum and the perinuclear cisterna indicating that these cells are actively synthesizing AcPase, probably due to a foregoing inductive event. Such stimulus could be the result of recent endocytosis of surface receptors in combination with antigen, antibody, or immune complexes and/or recent mitosis, or possibly some unrelated autophagic incident.

The authors report an unusual B-cell lymphoma that simulated hairy cell leukemia (HCL) not only clinically but also pathologically in peripheral blood, bone marrow, and lymph node specimens. A diagnosis of lymphoma could be made only after pathologic examination of the spleen, indicating that caution should be exercised in making a primary diagnosis of HCL on bone marrow examination. Morphologically this lymphoma resembled monocytoid B-cell lymphoma (MBCL) but the immunophenotype (monoclonal Ig-kappa +, Ia+, B4+, Leu-1+, LN-2+, lambda-, Bl-, LN-1- and CALLA-) and clinical findings were more consistent with mantle zone lymphoma (MZL). Because this case demonstrates features of both MBCL and MZL, the authors suggest that these two entities may have a common histogenesis.

We describe two childhood cases of a T-cell malignancy indistinguishable from nonendemic, idiopathic adult T-cell leukemia/lymphoma. The cases illustrate the heterogeneity of nonendemic, idiopathic ATLL, including the presence of mediastinal masses and more heterogeneous immunologic markers than endemic cases, and represent the two youngest cases of ATLL yet described. The differential diagnosis of nonendemic, idiopathic ATLL is discussed.

We studied the peanut agglutinin (PNA) staining patterns of histiocytosis X (H-X) (three cases, including one case of Letterer-Siwe disease) and well-characterized cases of malignant histiocytosis (two cases) and monoblastic leukemia (one case) in paraffin-embedded specimens. H-X cells showed staining identical to that of interdigitating reticulum cells, with unique paranuclear and cell surface deposits of reaction products. Thus, H-X cells were easily distinguished from benign macrophage-histiocytes, which show diffuse cytoplasmic staining, and malignant cells of malignant histiocytosis and monoblastic leukemia, which did not stain with PNA, PNA staining proved more useful than Leu-M1 antibody for this purpose, since neither interdigitating reticulum cells in ten reactive lymph nodes nor Langerhans'-type cells in three cases of H-X were stained with Leu-M1 antibody. These observations support a close relationship between H-X cells and interdigitating reticulum cells, and demonstrate the usefulness of PNA for the identification of these cells in paraffin-embedded material.

An immunoperoxidase technique was used to examine the distribution of lymphocyte subsets in bone marrow biopsies of 15 patients with neutropenia and seven non-neutropenic controls. The bone marrow of most patients and controls had similar distributions of immune effector cells characterized by a diffuse array of predominantly cytotoxic/suppressor T-cells and occasional nodular aggregates of helper T-cells. Cells displaying the natural killer cell marker HNK-1 were sparse in controls and most neutropenic patients. However, marked increases in marrow HNK-1 + cells were identified in four of the 15 patients. Three of these patients had diffuse HNK-1 + infiltrates associated with increased Leu 4+ (OKT-3+) T-cells while one had a nodular HNK-1+ infiltrate associated with small B-cell follicles. Each of these patients had clinical features similar to those described in the large granular lymphocyte (LGL) lymphocytosis (leukemia) syndrome, but only one of four demonstrated persistently increased numbers of LGLs in the peripheral blood. Thus, this study extends the association of neutropenia and increased numbers of cells with a T/NK phenotype to include patients whose bone marrow is the only demonstrable site of involvement. Since morphologic examination of the bone marrow could not identify the bone marrows with increased HNK-1+ cells, immunologic techniques are required to detect these cases.

This article describes the heterogeneity of post-thymic T-cell malignancies and correlates specific clinicopathologic entities with morphology and immunophenotype. Peripheral T-cell lymphomas are subclassified into six main histologic types with different prognoses. Special attention is given to the theme of proposed origin and spread of T-cell malignancies.

Reed-Sternberg and other Hodgkin's giant cells derived from involved spleens and lymph nodes of patients with Hodgkin's disease were examined for surface markers of T and B cells and macrophages. Attachment and phagocytosis of untreated (E) or sensitized (EA and EAC) sheep red blood cells and yeast by Reed-Sternberg cells did not occur. IgG frequently detected at the membrane of Reed-Sternberg cells was partially removed by incubation and washing at 37 degrees C. Fluorescence with a specific anti-T cell serum was not seen on Reed-Sternberg and other Hodgkin's giant cells lack most detectable normal human lymphoid cell markers, but do exhibit membrane bound immunoglobulin possibly of exogenous origin.

A hypothesis is proposed to explain the nature of the Reed-Sternberg (RS) cell and the unexpectedly common clinical associations between lymphomatoid papulosis, mycosis fungoides, and some types of Hodgkin's disease. The RS cell appears to be a lymphoblast arising from activated helper T cells. In lymphomatoid papulosis, a cutaneous eruption with histological features of Hodgkin's disease, there is a spectrum of activated helper T cells, including cerebriform cells and large transformed RS-like cells. Clonal expansion of cerebriform cells in lymphomatoid papulosis leads to mycosis fungoides. Similar expansion of large transformed cells results in Hodgkin's disease. Progressive transformation of T4-positive cerebriform cells to Ki-1-positive RS cells accounts for rare cases of coexistent mycosis fungoides and Hodgkin's disease. Confirmation of this hypothesis should focus attention on the family of human T-cell leukaemia/lymphoma viruses as possible aetiological agents in these helper-T-cell disorders.

This study was undertaken to explain the molecular basis for the diverse pathology and clinical behavior of postthymic T cell malignancies. Total cellular RNAs were extracted from four HTLV-1 positive and ten HTLV-1-negative T cell lymphomas and cell lines, and investigated for homology with cloned DNA probes specific for interleukin-2 (IL-2), IL-2 receptor (IL-2R), transforming growth factor beta (TGF-beta), platelet-derived growth factor (PDGF), and epidermal growth factor receptor (EGF-R). Tumor cells associated with clinically high grade HTLV-1-positive adult T cell leukemia (ATL) and large cell morphology (T immunoblastic lymphomas) were found to have higher levels of expression of IL-2 and TGF-beta genes than low grade T cell neoplasms (mycosis fungoides and Sezary's syndrome). High expression of IL-2R gene was restricted to Ki-1-positive lymphomas and to one ATL. Cell lines corresponding to the advanced stage of a cutaneous T cell lymphoma (CTCL) showed enhanced expression of PDGF. Therefore, high grade T cell malignancies had consistently elevated expression of growth factor/receptor (GF/R) genes. Expression of EGF-R was negligible in all T cell malignancies. An inverse relationship was found between the expression of T cell antigen receptor (differentiation antigen) and GF/R (activation antigen) genes, accounting for the frequent aberrant expression of T cell antigens in high grade T cell lymphomas. The results suggest that post-thymic T cell malignancies derived from activated T cells produce and secrete GF, conferring a growth advantage on neoplastic T cells, and correlating well with their histologic subtype and clinical behavior.

Angio-immunoblastic lymphadenopathy with dysproteinaemia (A.I.L.D.) is a lymphoproliferative disorder associated in one third of the patients with exposure to an allergen. In four patients with allergen-associated A.I.L.D. single cytotoxic drugs were ineffective but maintenance low-dose prednisone for 6--12 months induced complete remission for up to 13 years. In contrast, three patients with non-allergen-associated A.I.L.D. did not respond to prednisone but achieved remission with combination chemotherapy.

Lymphomatoid papulosis (LyP) is a papulonodular skin eruption characterized by spontaneous regression. In order to evaluate the role of host immune reactions in the phenomenon of regression, we analyzed skin biopsies from various stages of evolution in 16 patients with LyP using morphologic and immunoperoxidase techniques. Both rapidly regressing type A and more chronic type B lesions of LyP had modest perivascular and lesional infiltrates of small helper and suppressor T lymphocytes as well as histiocytes. Type A lesions in addition to numerous neutrophils and eosinophils had a more extensive lymphocytic infiltrate than type B lesions. Natural killer cells were not prominent in the skin lesions, although several patients had increased numbers of circulating natural killer cells. Our results demonstrate a correlation between the inflammatory components found in skin biopsies from patients with LyP and clinical behavior and suggest that cell-mediated immune reactions may play a role in the spontaneous regression of LyP.

The L-428 cell line derived from nodular sclerosing Hodgkin's disease was verified to be a human female cell line with surface marker and morphologic characteristics similar to native Hodgkin's cells. Single cells were cloned and subcloned twice to determine the characteristics of the clonogenic L-428 Hodgkin's cell (resulting in a 10% cloning efficiency). Both mononuclear L-428 cells and classical Reed-Sternberg cells arose from solitary cells. The clonogenic cell was the mononuclear Hodgkin's cell, although small abortive colonies sometimes arose from classical binucleate Reed-Sternberg cells. Cytogenetic and phenotypic analysis supported the clonality of three subclones and indicated, among many findings, consistent abnormalities of the long arm of chromosome 7 (beta-chain of the T cell receptor) and 14q32 (Ig heavy chain). Distinctive abnormalities of cytogenetics, phenotyping and transforming growth factor-beta production were exhibited for each clone as well. These observations demonstrate the relationship of the continuum of malignant mononuclear and multinuclear Reed-Sternberg cells in this cell culture from nodular sclerosing Hodgkin's disease and suggest that a similar relationship exists in native Hodgkin's disease tissue. These observations also support the theory of clonality in Hodgkin's disease and suggest that in vivo contiguous metastasis in the L-428 Hodgkin's disease patient was most likely accomplished by a Ki-1 positive small mononuclear cell.

We studied Reed-Sternberg cells from 14 patients with Hodgkin's disease to learn whether they had monoclonal immunoglobulin synthesized by the cell or polyclonal immunoglobulin of external origin. Double-label immunofluorescence with F(ab')2 anti-serums to human light chains showed that cytoplasmic immunoglobulin of individual Reed-Sternberg cells is always polyclonal and usually associated with membrane-bound immunoglobulin of the same type. The predominant immunoglobulin was IgG; in one case IgM was also present. In vitro studies confirmed the internalization of exogenous IgG and phagocytosis of immune complexes by viable Reed-Sternberg cells. Their exclusion of trypan blue dye and lack of albumin and fibrinogen suggests relatively specific uptake of immunoglobulin, mediated by the Fc receptor or antigen (or antigens) associated with Hodgkin's disease at the cell membrane. Our studies support other recent evidence that the Reed-Sternberg cell is derived from a macrophage.

A patient with a long history of celiac sprue developed a pleomorphic intestinal lymphoma. Cell suspensions of tumor cells and immunoperoxidase labeling of cell surface antigens in frozen tissue sections clearly showed the lymphoma to be of T-cell origin, despite the presence of other nontumor cells bearing enzyme markers of histiocytes. These findings may have important implications on the cellular origin of lymphomas developing in patients with celiac sprue.

Nodular sclerosing Hodgkin's disease is characterized by dense collagen fibrosis. Although transforming growth factor-beta (TGF-beta) is an important bifunctional growth factor for fibroblasts and is stored and released by many cells, it requires acidification to pH 2.0-3.0 before it becomes a biologically active growth factor. We show here that the L-428 Hodgkin's cell releases a high molecular weight TGF that competes for the TGF-beta cell membrane receptor but not the TGF-alpha receptor. This growth factor is most active at physiologic pH and is 97% inactivated by acidification. Hodgkin's TGF is also inactivated by proteases and can be preserved by protease inhibitors. The Hodgkin's TGF can be separated from an autocrine growth factor using either column chromatography or electroelution from gels and is shown to have a molecular weight of approximately 350,000. Incubation of the Hodgkin's TGF in SDS releases a 25,000-D protein with reduced biological activity but which cross-reacts with anti-TGF-beta IgG. We propose that L-428 nodular sclerosing Hodgkin's disease fibrosis is mediated by a potent high molecular weight TGF-beta which, unlike TGF-beta characterized to date, is secreted in a form most active at physiologic pH.

One hundred four cases of malignant lymphomas, including 90 cases of non-Hodgkin's lymphoma, 5 cases of histiocytic malignancy, and 9 cases of Hodgkin's disease were analyzed pathologically and immunologically using a panel of monoclonal and conventional antibodies for T-, B-, histiocyte, and Hodgkin's neoplastic cells. Our results revealed a high frequency of T-cell lymphoma (42.3%), a low percentage of follicular lymphoma (10.5%), and Hodgkin's disease (8.7%) in Taiwan. More than half of the malignant lymphomas belonged to the high-risk unfavorable group. Peripheral T-cell lymphomas (33 cases) showed characteristic clinical and histologic features, which can sometimes be confused with Hodgkin's disease. Monoclonal antibodies Leu-M1 and 2H9 were an important aid for their differential diagnosis. Five of the 33 peripheral T-cell lymphomas were positive for antibody to adult T-cell lymphoma/leukemia (ATL) virus associated antigen (ATLA). Four patients were from the northeast coast of Taiwan, I-Lan county. Five (4.8%) were diagnosed as true histiocytic malignancies, including two true histiocytic lymphoma and three malignant histiocytosis. Two cases each of large cell lymphoma and immunoblastic lymphoma showed no identifiable marker expression. The distribution of lymphoproliferative disorders in Taiwan is similar to that in Japan but much different from western countries.

Pure red cell aplasia (PRCA) results from the failure of erythrocyte differentiation and may respond to immunosuppressive therapies. We have treated nine patients with PRCA refractory to steroids and/or cyclophosphamide with anti-thymocyte globulin (ATG). Six patients had normal numbers of erythroid bursts (from erythroid burst-forming units) or erythroid colonies (from erythroid colony-forming units) detectable in vitro, and all responded to therapy with ATG. In vitro studies suggested T-cell inhibition of erythropoiesis in four of these six patients and humorally mediated erythroid suppression in one. In three individuals, virtually no erythroid progenitors were detected in marrow culture. None of these patients responded to ATG. Myelofibrosis, 5q- chromosomal abnormality, or the subsequent development of thrombocytopenia in these individuals suggested that PRCA resulted from an intrinsic stem cell disorder. Our studies demonstrate that ATG is effective therapy for PRCA, and it may be especially useful in children or other patients in whom alkylating agents are not appropriate. We also confirm that erythroid growth in marrow culture predicts those patients who will respond to ATG or other immunosuppressive therapies.

The authors analyzed 50 cases of Hodgkin's disease (HD) with a panel of antibodies which detect B-cell and T-cell specific markers and activation antigens using a sensitive immunocytochemical technique and paraformaldehyde-lysine-periodate (PLP) fixed-frozen tissues. In 60% of cases either T-cell or B-cell specific antigens were detected on Reed-Sternberg (RS) cells. Most T-cell cases were of nodular sclerosing (NS) and mixed cellularity (MC) type (65% and 30%, respectively) and most B-cell cases were either of NS or lymphocyte predominant (LP) type (55% and 45%, respectively). Leukocyte common antigen (LCA) was usually negative on RS cells in NS, but was present in approximately 50% of the cases of MC and LP types. Almost all cases were positive for the CD30 antigen (Ki-1). Most cases were also positive for CD15 (LeuM1) with the exception of the LP type. Activation antigens (Ia, CD25, T9) were expressed in a high proportion of cases regardless of subtype. The results suggest that most cases of HD are histogenetically derived from activated T-cells or B-cells.

We attempted to determine the reliability of surface markers in distinguishing 21 small round cell tumors from lymphoid malignancies. Using immunofluorescence on tumor cell suspensions and immunoperoxidase on fresh frozen sections, we found that specimens of neuroblastoma (n = 7), rhabdomyosarcoma (n = 7), Ewing's tumor (n = 5), and two unclassified small round cell tumors all lacked human HLA-DR antigens. Each of eight tumors tested also lacked common leukocyte antigen (T200). In each of 13 cases studied, neither polyvalent surface immunoglobulin (sIg) nor receptors for sheep erythrocytes (E), complement (EAC), or the Fc portion of IgG immunoglobulin (EA) were found. Conversely, we found HLA-DR and/or T200 antigens, usually one or more receptors for E, EAC, or EA, and not infrequently, monoclonal sIg on malignant cells in each of 42 cases of lymphocytic lymphoma and leukemia. We conclude that study of surface DR and T200 antigens, sIg, and receptors for E, EAC, and EA aids the differential diagnosis of small round cell tumors from lymphocytic lymphoma and leukemia.

We performed splenectomy in four patients with severe neutropenia (less than 0.5 X 10(9)/l), recurrent infections, and splenomegaly associated with large granular lymphocyte leukaemia. Prior to splenectomy, elevated levels of neutrophil-reactive IgG were detected in sera of all three patients tested. In all patients, enlargement of the spleen was due to a characteristic lymphoid infiltration of red pulp cords. Splenectomy resulted in an increased neutrophil count greater than 0.5 X 10(9)/l in all patients; this response was sustained in two patients who benefited clinically by a dramatic reduction in frequency of infections. Poor clinical response was associated with elevated levels of antineutrophil antibody post-splenectomy. All four patients had an increase in number of circulating large granular lymphocytes post-splenectomy; one patient who had attained a sustained neutrophil response died of an accelerated lymphoproliferative disorder 19 months post-splenectomy. We conclude that splenectomy may be of value in correcting severe neutropenia and reducing infections in some patients with large granular lymphocyte leukaemia. However, splenectomy appeared to be of no value in treatment of the underlying lymphoproliferative disorder.

The authors studied the occurrence of peanut agglutinin (PNA)-binding cells in paraffin-embedded specimens of 145 patients with Hodgkin's disease (HD). The staining reaction of lymphocytes was consistently negative. A positive staining reaction was observed in two types of cells: macrophage-histiocytes (M-H), and Reed-Sternberg (R-S) cells and their variants. Diffuse or globular cytoplasmic staining was found in M-H, which was easily distinguished from a unique "cell surface and cytoplasmic" staining pattern of R-S and related cells. Thus defined, M-H were numerous in lymphocyte depletion and mixed cellularity, less common in lymphocyte predominance, and least frequent in the nodular sclerosis type. Numerous M-H correlated with B-symptoms and a poor response to therapy. Among the asymptomatic patients with localized disease at presentation, the presence of large numbers of M-H was associated with a high incidence of relapse within 2 years of therapy. These findings suggest that the number of non-neoplastic M-H in HD may be an important determinant in the clinical presentation and course of disease. Peanut agglutinin staining may be useful for the detection of M-H in routine diagnosis and classification of HD, which has not been feasible by conventional methods.