H Uchino

Effect of adult T-cell leukemia cells on pokeweed mitogen-induced normal B-cell differentiation

Abstract Leukemic T cells from Japanese patients with adult T-cell leukemia were studied for their effects on pokeweed mitogen-induced peripheral blood lymphocyte differentiation into immunoglobulin-producing cells. Allogeneic peripheral blood lymphocytes, normal T cells, nonlymphoid leukemia cells, lymphoid leukemia cells other than adult T-cell leukemia cells, and long-term T-cell lines did not suppress normal B-cell differentiation in coculture experiments. Leukemic T cells from three of six patients with adult T-cell leukemia showed a marked suppressive effect. Supernatant fluids obtained from a short-term (48-hr) culture of leukemic T cells from one of these three patients also showed a marked suppressive effect on B-cell differentiation at a final dilution of 1 5 or less when added at the beginning of the culture (on Day 0) or on Day 1. Supernatant fluids, however, did not manifest suppressor activity when added on Day 3 or later, or when heated at 56°C for 30 min before addition.

Selective IgA deficiency in Japanese blood donors: frequency and statistical analysis.

Abstract. The incidence of selective IgA deficiency (SIgAD) was determined in a healthy adult population of 222,597 Japanese volunteer blood donors. Of the blood donors screened, only 0.007% (1:14,840) were found to be IgA‐deficient (less than 10 mg/dl) by means of the double diffusion method, while 0.005% (1:18,500) were less than 5 mg/dl, and 0.003% (1:31,800) were less than 1 mg/dl by means of the single radial immunodiffusion method. Statistical analysis of the results clearly showed that the incidence of SIgAD in Japanese blood donors is very much lower than that in blood donors of European ancestry. The Japanese population may occupy a unique position in the ethnical peculiarities. Anti‐IgA antibodies were found in 3 (25.0%) of 12 IgA‐deficient blood donors whose IgA levels were less than 5 mg/dl, a prevalence rate comparable to that in donors of European ancestry. Although it is difficult to develop a suitable file of IgA‐deficient donors in Japan, the establishment of a Rare Donor Registry System on IgA deficiency is a matter of urgency.

Suppressive activity of some leukemic T cells from adult patients in Japan

Abstract When the stimulatory capacity of leukemic lymphocytes was investigated in mixed lymphocyte culture (MLC) with peripheral blood lymphocytes from healthy individuals, some leukemic T cells from Japanese adult patients reduced background proliferation of normal lymphocytes. These leukemic T cells were effective in reducing normal lymphocyte response to phytohemagglutinin (PHA) or to lymphoblastoid cell lines with B-cell properties (B-LCL) in MLC, when added as attending cells, and the activity was mitomycin-C resistant. The culture supernatants also reduced normal lymphocyte response to B-LCL, but failed to suppress markedly the response to PHA.

Report of a patient with POEMS/takatsuki/crow-fukase syndrome associated with focal spinal pachymeningeal amyloidosis

The authors present the results of an autopsy of a 67‐year‐old Japanese man with POEMS/Takatsuki/Crow‐Fukase syndrome (P/T/CFS) diagnosed in 1972. Each component of the syndrome was gradually recognized after the resection of lumbar vertebral solitary plasma‐cytoma in 1967. The patient died in 1989 of generalized infection and renal failure. Autopsy revealed in the vertebral canal between the fifth and seventh thoracic vertebrae dorsal pachymeningeal fibrosis, with prominent amyloid deposition that oppressed the spinal cord. This condition was consistent with the final neurologic manifestation of the patient, bilateral motor and sensory disturbance below the sixth thoracic level. Myelopathy remained clinically unnoticed because neurologic disturbance had begun as peripheral polyneuropathy. Normo‐cellular marrow with heterogeneously scattered lambda light chain‐positive plasma cells and degeneration of the myelinated fibers of sciatic nerve also were observed. This is the first report of focal spinal amyloidosis associated with P/T/CFS. Cancer 1992; 70:882–886.

Stromal Cell-dependent Growth of Leukemic Cells from Murine Erythroblastic Leukemia

Transplantable erythroblastic leukemia was induced by 300‐rad irradiation of C3H mice. Conditions for in vitro growth of the leukemic cells were studied. None of interleukin‐3, granulocyte/macrophage colony‐stimulating factor and erythropoietin could support the growth of the cells in vitro. In contrast, the leukemic cells grew into a stroma‐dependent cell line, ELM‐D, in close contact with the stromal cell layer of 900‐rad‐irradiated long‐term bone marrow culture. A stroma‐independent cell line, termed ELM‐I‐1, was further established from the non‐adherent population in the co‐culture of the leukemic cells, ELM‐D, with stromal cells. Reverse transcriptase activity was not detectable in ELM‐D or ELM‐I‐1 cells. Studies on binding and cross‐linking of 125I‐erythropoietin showed that ELM‐I‐1 cells had erythropoietin receptors, and two major radiolabeled protein products with molecular weights of 120 kDa and 140 kDa were detected on sodium dodecyl sulfate/polyacrylamide gel electrophoresis under reducing conditions.

A Novel B Cell Line Established from Ki-1-positive Diffuse Large Cell Lymphoma

A novel cell line, designated KIS‐1, was established from a patient with Ki‐1‐positive diffuse large cell lymphoma. Multiple phenotypic analysis of the KIS‐1 cells was carried out with a total of 22 monoclonal antibodies defining hematopoietic cell subsets and lineages. The KIS‐1 cells were positive for Ki‐1, B4, HLA‐DR, and 2D1 (common leucocyte) antigens, but were negative for the antigens reportedly specific for T cells, natural killer cells, granulocytes, monocytes, interdigitating reticulum cells and dendritic reticulum cells. The genomic analysis of the KIS‐1 cells showed not only the rearrangement of JH and Jk genes but also the probable rearrangement of Cγ genes. Moreover, the cells produced immunoglobulin γ chains. Thus, KIS‐1 was considered to be of B‐cell lineage. The lymphoma‐cell derivation of KIS‐1 was based on the following facts. The cytochemical, immunologic, cytogenetic properties and the results of the molecular genomic analysis in the KIS‐1 cells were essentially the same as those of the original tumor cells, and the KIS‐1 cells were negative for Epstein‐Barr virus‐associated nuclear antigen. KIS‐1 is the only known B‐cell line derived from Ki‐1‐positive diffuse large cell lymphoma, and should be useful for defining the biological implications of Ki‐1 antigen.

SERUM LEVELS OF SOLUBLE INTERLEUKIN 2 RECEPTOR IN PATIENTS WITH NON-HAEMATOLOGICAL DISORDERS

Recently Pizzolo et d (1987) reported the elevation of soluble Interleukii 2 Receptor (sIL 2R) levels in haematological disorders, i.e. hairy cell leukaemia, Hodgkin’s disease, nonHodgkin’s lymphoma, B-chronic lymphocytic leukaemia, etc. They described that the increase of sIL 2R levels is not diseasespecific but it can be clinically useful as a index of disease activity in a number of haematological disorders. They found no obvious increase of sIL 2R levels in most non-haematological malignancies. We have examined serum sIL 2R levels in a number of nonhaematological disorders (Table I). sIL 2R values were determined using the enzyme-linked immunosorbent assay (Cellfreem interleukin-2 receptor kit, T cell Sciences, Inc., Cambridge, Mass.), based on the use of two monoclonal antibodies (2R1.2 and 7G7/B6) developed against two different epitopes of the IL 2R molecules (MacKeen et al, 1986). Serum levels of sIL 2R were expressed in Units/ml (U/ml) relative to a set of standards supplied with the test kit which Pizzolo’s group also used (Chilosi et al. 1987). In 29 patients with chronic renal failure, the levels of sIL 2R were significantly higher than those seen in normal controls. These patients had received the haemodialysis twice or three times a week and had high levels of urea nitrogen in the blood (BUN= 80.9 f 22.4 mg/dl, meanf SD) before the haemodialysis. No significant decrease of sIL 2R values was found after the haemodialysis. Sixteen patients with non-haematological malignancies, including seven gastric cancer, three colon cancer, three lung cancer and others, also showed high serum levels of sIL 2R. Furthermore, elevated serum sIL 2R values were found in infection. Sixteen patients with infectious disease, including eight viral disease and eight bacterial disease, revealed elevated sIL 2R values. The very high sIL 2R values (up to 50000 U/ml) as observed in adult T-cell leukaemia (Yasuda et al, 1988) or hairy cell leukaemia (Pizzolo et aJ, 198 7) were not detected in our series of the patients with non-haematological disorders. However, in the remission state or the smouldering cases of haematological disorders (Table I), their levels of sIL 2R are very close to those seen in our cases. Our &dings suggest that sIL 2R levels may vary not only with disease activity of haematological disorders but also with the renal function and the presence of infection and non-haematological malignancies. Hence, when serum sIL 2R values might be used as a index of disease activity of haematological disorders, one should rule out the influences from these factors.

Vacuolated plasma cell: ultrastructural distribution of acid-phosphatase and intracellular immunoglobulin

In two cases of vacuolated plasma cells, one of which was associated with primary macroglobulinemia and the other was with kappa-chain Bence Jones multiple myeloma, we examined the immunopathological features of the vacuoles in order to know whether the Ig-secreting system or the lysosomal system is of importance in the process of vacuole formation. Immunofluorescent studies detected no Ig in the vacuoles. Detection of intracellular Ig by immunoelectron microscopic technique revealed that Ig was localized only in a small portion of the vacuoles but not in most vacuoles. Even when Ig was included in the vacuoles, only the contents of the vacuoles were positive for Ig but their demarcating membrane was negative for Ig. In contrast, electron microscopic studies of acid phosphatase activity revealed the presence of its activity in all vacuoles. These findings suggest that the lysosomal system but not the Ig-secreting system may play a major role in vacuolation of these myeloma cells.

A New Hypothesis on the Cellular Origin of Reed-Sternberg and Hodgkin Cells Based on the Immunological and Molecular Genetic Analysis of the KM-H2 Line

Previously we reported a novel cell line, KM-H2, established from the pleural effusion of a patient who was initially diagnosed as having Hodgkin’s of mixed cellular type (Kamesaki et al. 1986). We describe here the results of further investigation of this line (ultrastructural, immunological, and molecular genetic studies) and discuss its cellular origin. Based on these analyses of the KM-H2 line, as well as those of other cell lines derived from Reed-Sternberg and Hodgkin cells (e.g., L428, L540), we propose a new hypothesis for the cellular origin of Hodgkin’s disease.

Sandwich radioimmunolabeling for the study of surface properties of bone marrow lymphocytes.

A modification of sandwich radioautographic method was applied to the study of surface immunoglobulin and/or specific antigens on small lymphocytes in mouse and human bone marrow. After incubation of marrow cell suspensions at 37 degrees C, cells were reacted at 0 degree C for 30 min with graded dilutions of rabbit anti-mouse or anti-human immunoglobulin followed by further reaction with a sheep anti-rabbit immunoglobulin labeled with 125I. Detectable surface immunoglobulin was demonstrated in approximately one-third of mouse marrow lymphocytes and 20-25% of human marrow lymphocytes. The densities of surface immunoglobulin as assessed by grain counts on individual labeled lymphocytes tended to be lower in the marrow than in spleen or peripheral blood. When the same rabbit antiserum was used to compared the sensitivity of the sandwich method with that of the direct radioautography, the former was found sufficiently sensitive to give a plateau level of labeling without seriously increasing background grains. The advantages of the method are discussed with reference to studies on T and B cells specific antigens on human bone marrow lymphocytes.