Lymphoid Leukemia Cells Research Articles

"Target" cells, cellular phenotypes, and lineage fidelity in human leukaemia.

A detailed analysis of lymphoid leukaemia cells with a panel of monoclonal antibodies plus other markers indicates a striking conservation of phenotype that closely mimics that of normal lymphoid precursor or progenitor cells and reflects the imposition of maturation arrest. The composite phenotypes detected show some minimal deviations or asynchronies (with respect to maturation "stages") but are faithful with respect to cell lineage. These observations are suggested to shed light on the available "target" populations for lymphoid malignancy in humans and challenge the view that dedifferentiation or qualitatively aberrant gene expression is common and important in malignancy. They are further taken to support the view that normal gene products that regulate or couple proliferation and maturation are critically involved in the successive clonal selection events underlying the evolution of leukaemia. No qualitatively unique and consistent leukaemia "markers" may exist or be required. The only exception to this rule are the nonrandom chromosome changes, which, it is suggested, involve genetic loci critical to the regulation of growth and maturation in particular normal cell types.

Involvement of human membrane-associated complement components in the rosette formation between Marmoset red blood cells and human leukocytes

The majority of human monocytes, a subpopulation of B lymphocytes, and all B lymphoblastoid cell lines (B-LCL) tested but one form rosettes with Marmoset red blood cells (MaRBC). None of the human peripheral T cells, T-LCL, and B chronic lymphoid leukemia cells (B-CLL) used bind to MaRBC. The binding could not be correlated with any membrane markers or antigens present on cultured cells or peripheral blood leukocytes (PBL). Blocking of the rosette formation by preincubation of MaRBC with purified human complement (C) components and cobra venom or by pretreatment of leukocytes and cultured cells with antisera to human C components suggested that membrane-associated C components present on PBL or B-LCL are involved in the binding to MaRBC.

Hematopoietic cells transformed in vitro by REV T avian reticuloendotheliosis virus express characteristics of very immature lymphoid cells

REV T (avian reticuloendotheliosis virus) is a replication defective acute leukemia virus (J. D, Hoelzer, R. B. Franklin, and H, R. Bose, 1979, Virology 93, 20–30) possessing a transformation-specific sequence designated rel. Hematopoietic cells transformed by REV T in vitro expressed cell surface markers of immature B lymphocytes. Some clones of transformed cells also expressed cell surface determinants of immature T lymphocytes. An antiserum specific for REV T-transformed cells reacted with the cell surface of lymphoid leukemia cells of both the B- and T-cell lineage as well as with a subpopulation of cells in normal spleen, thymus, and bone marrow. REV T-transformed cells did not express surface or cytoplasmic immunoglobulin chains nor terminal deoxynucleotidyl transferase, nor were they susceptible to infectious bursal disease virus. As expected, they were also negative for the expression of all erythroid and myeloid differentiation parameters tested.

Manifold expression of new cellular genes in human lymphoid neoplasia.

Previous studies performed in our laboratory have shown that a minor single-stranded DNA component (ssDNA) isolated from the bulk of double-stranded nuclear DNA (nDNA) of human, murine, or avian cells consisted mainly of active transcription sites. In the present work, ssDNA isolated from human lymphoid cells, either malignant or not, amounted to 1.2-1.4% of the total nDNA. This was labeled with 125I and utilized as a probe in RNA-driven iterative hybridization and cross-hybridization assays. In all cases of neoplasia studied, the same subfraction of ssDNA (equivalent to 10-12% of the total ssDNA) from malignant cells could be hybridized with cytoplasmic RNA from malignant cells, whatever their origin: acute lymphoid leukemia cells collected from blood of leukemic patients, local lymphosarcomas, or cells from two established strains grown in suspension, one Epstein-Barr virus positive (Raji strain), the other Epstein--Barr virus negative (Ramos strain). The majority of these tumor-specific DNA sequences could be reassociated with unique-copy DNA from both normal and malignant cells and were not expressed in normal lymphoid cells, including those of an established strain immortalized by Epstein--Barr virus and multiplied in culture. It is concluded that a great number of new transcription units, in the range of 2500-3000 per cell genome, were activated after spontaneous malignant transformation of lymphoid cells.

The contribution of scanning electron microscopy in haematology: its role in defining leucocyte and erythrocyte disorders.

This report reviews the contribution of scanning electron microscopy (SEM) in haematology. Important observations regarding red cell shape and deformities are referred to and contributions in the definition and classification erythrocyte disorders are stressed. In this field there is no doubt that SEM has contributed much to the three-dimensional visualization of RBC disorders. As far as leucocyte pathology is concerned the situation is less clear. SEM has contributed much to current knowledge and understanding of the surface properties of white blood cells. Normal leucocytes have different surface features and can be distinguished under the SEM. However, some overlap does occur, making individual distinction on the basis of surface architecture alone extremely difficult. The difficulties in this regard are discussed in this review and factors influencing the variability of surface microprojections are reviewed briefly. Leukaemic cells of different origins may also be distinguished under the SEM. However, while "hairy' cells have typical surface features and non-lymphoid leukaemic cells, in particular monocytes, are readily distinguished from lymphoid leukaemic cells, there is much overlap of surface topography. Undifferentiated cells and early myeloblasts and lymphoblasts have similar surface features and cannot be distinguished under the SEM. While SEM adds a valuable third dimension to morphology and ultrastructure, it cannot be used alone in the definition of difficult cases of acute leukaemia.

Nature of leukemia-associated antigenicity of dinitrophenylated human lymphocytes and lymphoblasts

Using dinitrophenylated human lymphocytes and phytohaemagglutinin-stimulated human lymphoblasts as antigens, antibodies were produced in rabbits. The immunological reactivities of the antisera so produced were tested against various types of leukemic cells after absorbing the sera with pooled normal leukocytes. Both the sera showed reactivity with all types of leukemic cells and no specific affinity for lymphoid leukemic cells was seen. This may suggest the presence of some common antigens on all types of leukemic cells or that dinitrophenylation brings about similar changes on all types of normal leukocytes.

Evidence for the cellular origin of Gross virus-induced leukemia in the rat: description of a unique LDH isozyme sub-band in leukemic lymphoid cells and lymphohemopoietic precursor cells.

Neoplastic thymocytes from rat thymic lymphoma-leukemias induced by the rat-adapted Gross-leukemia virus (RAGV) were analyzed for a variety of differentiation markers to define their differentiation state and possible cellular origin. A majority of thymocytes from leukemic rats had the phenotypic characteristics of subcapsular cortical thymocytes that are the most ancestral of the thymocytes. These cells exhibited readily detectable levels of Thy-1 and histocompatibility antigens on their surfaces, they contained terminal deoxynucleotidyl transferase (TdT) and they contained low adenosine deaminase (ADA) and high purine nucleoside phosphorylase (PNP) specific activity. The leukemic thymocytes also contained a sub-band of the LDH-5 isozyme (LDH-5') that was not detected in normal thymocytes but that was present in lymphocyte-rich fractions of postnatal bone marrow, fetal and prepubertal spleen, and fetal and neonatal liver. The tissue distribution and ontogeny of LDH-5'-containing cells is similar to prethymic TdT+ cells in the rat and both TdT and LDH-5' are enriched in a subset of bone marrow "null" cells. These results suggest that TdT+ thymocyte progenitors or their precursors are the targets of leukemic transformation of RAGV.

Destruction of erythroleukemia, myelocytic leukemia and Burkitt lymphoma cells by photoactivated protoporphyrin.

The effect of protoporphyrin on erythroid, myeloid and lymphoid leukemic cells and their destruction induced by the photoactivated porphyrin was studied. Friend erythroleukemic cells (FL) and myelocytic leukemic cells (ML) accumulated protoporphyrin in a cap or patch-like pattern observed by fluorescence microscopy. Photoactivated protoporphyrin induced the appearance of "holes" on the cell membrane demonstrated by scanning electron microscopy. On the other hand, Burkitt lymphoma (BL) and mastocytoma (MS) cells accumulated porphyrin intracellularly around the nuclear envelope and as circular profiles, respectively. Photoactivated protoporphyrin induced development of multiple blebs on the cell membrane, and even complete cell destruction. Cytotoxicity of protoporphyrin at short-term incubation periods was determined by [3H]thymidine and [3H]uridine incorporation. Protoporphyrin, unexposed to light, reduced the incorporation of both precursors only to a moderate extent. On the other hand, porphyrin-treated cells exposed to light showed complete inhibition of RNA and DNA synthesis. Long-term exposure of ML and BL cells to porphyrin in the dark induced a nearly 50% inhibition of RNA and DNA synthesis. Although the cytotoxic effect of protoporphyrin in the dark was lower than that of photoactivated porphyrin, this may possess a potential activity in vivo even without illumination.

Lectin staining of carbohydrates of haemic cells. II. The cells of normal lymphoid origin, of lymphatic leukaemias and related diseases.

Normal and leukaemic lymphoid cells, both human and murine, were stained for specific carbohydrates with three fluorescein-labelled lectins: Aprotinin for sialyl (or uronyl) groups: Ricinus agglutinin for galactosyl groups; and Concanavalin A for mannosyl (or glucosyl) groups. The method gives permanent preparations of sections from methanol fixed, paraffin embedded tissues, from blood and bone marrow films or touch preparations of lymph nodes that were methanol fixed. Whereas normal lymphocytes and lymphoblasts reacted strongly for sialyl groups, lymphoblasts of acute lymphoblastic leukaemia and lymphocytes of chronic lymphocytic leukaemia gave a much weaker reaction. The same was the case of the lymphocytes of the Sézary variant and the lymphocytes of macroglobulinaemia. The fine processes of the cells of hairy cell leukaemia stained well for sialyl groups. No obvious differences were detected between normal monocytes and the cells of monocytic leukaemia, nor between normal plasma cells and those of myeloma.

Adenosine Triphosphatases on Plasma Membrane and Membrane Surrounding Lipid Vacuoles in L1210 Lymphoid Leukemic Cells

The activity of adenosine triphosphatases in L1210 lymphoid leukemic cells was demonstrated by ultracytochemical methods. The activity of Na+, K+-ATPase was found only in the plasma membrane, but the activity of Mg2+-ATPase and Ca2+-ATPase was found in both the plasma membrane and in the membrane surrounding lipid vacuoles in the cytoplasm. The activity of Ca2+-ATPase predominated on the periphery of the lipid vacuoles. The localization of these enzymes is discussed in reference to its possible significance in transformed cells.

Cytochemical visualization of calcium binding to the plasma membrane of L-1210 lymphoid leukaemic cells1)

In L-1210 lymphoid leukaemic cells fixed in glutardialdehyde supplemented with CaCl2, calcium-dependent electron dense deposits were detected. The deposits were observed on the extra-cellular surface and were nonfrequent. Abundant deposits were also found at the periphery of lipid globules. The formation of deposits and the possible physiological significance of these findings are discussed.

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

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.

Membrane Marker and Cell Separation Studies in Ph1-positive Leukemia

AbstractFrom 32 patients with Ph1-positive leukemia whose cells reacted with a specific antiserum made against non-T, non-B acute lymphoid leukemia (ALL) cells, five typical patients were studied in detail. Three of these had had Ph1-positive chronic myeloid leukemia (CML) and later developed acute blast crisis with lymphoid involvement. The other two patients presented with acute leukemia with a mixture of myeloid and lymphoid blasts. In all five cases lymphoid blasts expressed the same phenotype as Ph1-negative non-T, non-B ALL: they reacted with anti-ALL and anti-p28,33 (anti-la) sera but lacked differentiation markers of thymocytes, B lymphocytes, and myeloid cells. In these cell populations high levels of terminal transferase enzyme activity were detected. Blast cells reacting with anti-ALL serum (ALL+) were separated from unreactive cells (ALL-) on a fluorescence-activated cell sorter. ALL+ cells had the morphology of ALL blasts, while ALL- cells appeared to be myeloid. These studies also suggest but do not prove that both ALL+ (lymphoid) and myeloid populations carried the Ph1 chromosome. The implication of these findings is that at least some Ph1-positive leukemias may arise from a prelymphoid, premyeloid progenitor and during blast crisis a variable proportion of blasts may retain their essentially stem cell-like characteristics. Since patients with lymphoid involvement sometimes respond to therapy designed for ALL, the early diagnosis is clinically important. Assays for ALL antigen and for terminal transferase have an important role in the diagnosis of lymphoid blast crisis, particularly if the lymphoblasts are present in the company of myeloid cells.

ChemInform Abstract: INHIBITORS OF POLYAMINE BIOSYNTHESIS. V‐ IN VIVO EFFECTS OF α‐METHYL‐(.+‐.)‐ORNITHINE AND α‐METHYL‐(.+‐.)‐ORNITHINE TERT‐BUTYL ESTER

AbstractDie Wirkung der Titelverbindungen auf die Lebensdauer und den Polyamidspiegel von Mäusen, die mit L‐1 210 lxmgh ‐ tischen Leukämie‐Zellen behandelt worden sind, wird untersucht.

1,2,4-Triazole amino nucleosides. 1-.beta.-D-3'-amino-3'-deoxyribofuranosyl-1,2,4-triazole-3-carboxamide and related nucleosides

The synthesis of 1-beta-D-3'-amino-3'-deoxyribofuranosyl-1,2,4-triazole-3-carboxamide--the 3'-amino analogue of ribavirin--and five related nucleoside analogues is described. Each analogue exhibited LD50 concentrations greater than 100 microgram/mL against P-388 mouse lymphoid leukemia cells in tissue culture. Antiviral testing indicated that none of the compounds exhibited significant activity.

Inhibitors of polyamine biosynthesis V: In vivo effects of alpha-methyl-(+/-)-ornithine and alpha-methyl-(+/-)-ornithine tert-butyl ester.

Abstract The effects of a-methyl-(±)-ornithine and its tert -butyl ester on the survival time and the levels of polyamines in spleen tissue of mice inoculated intraperitoneally with L-1210 lymphoid leukemic cells were studied in vivo . These compounds were administered at doses of 100–300 mg/kg from Day 1 of tumor inoculation until death. This treatment neither significantly increased survival time nor altered the increases in polyamine levels normally observed during tumor growth. A method was developed to determine polyamine levels in spleen tissue. The polyamines in the tissue extracts were converted to their 5-(dimethylamino)-1-naphthalenesulfonyl derivatives, and these derivatives were separated by TLC followed by high-pressure liquid chromatography on a bonded phase microparticle column. The in vivo metabolism of 1- 14 C- α -methyl-(±)-ornithim was studied and compared to that of 1- 14 C-(±)-ornithine. 1- 14 C- α -Methyl-(±)-ornithine was rapidly excreted after administration to mice inoculated with L-1210 leukemic cells. Within 24 hr after injection, over 70% of the radioactive dose was found in the urine, primarily as the unchanged compound. Less than 0.1% of the administered dose appeared as 14 C-labeled carbon dioxide during the 24 hr following injection. In contrast, 34% of the administered dose of 1- 14 C-(±)-ornithine was metabolized to labeled carbon dioxide and 28% was excreted in urine during the 24 hr following injection.

Possible effects of non-HLA antibodies in common typing sera on HLA antigen frequency data in leukemia

Selective adsorption of several common "monospecific" HLA typing sera with HLA typed platelets, purified B lymphocytes, cultured lymphoid cells, or lymphocytes from patients with active chronic lymphocytic leukemia demonstrated that many of these sera contain antibodies to non-HLA antigens. Antibodies were detected to antigens present on peripheral blood B-lymphocytes, cultured lymphoid cells and leukemic cells from patients with both myelocytic and lymphocytic forms of leukemia but absent from T-lymphocytes and platelets. Since these kinds of antibodies appear to be present in a large proportion of common HLA typing sera, caution should be used in interpreting all data related to HLA antigen expression in leukemia.

Acute Lymphoblastic Leukemia (ALL) Antigens Detected with Antisera to E Rosette-Forming and Non-E Rosette-Forming All Blasts

Abstract Based on the presence or absence of erythrocyte receptors(E) a T cell marker, acute lymphocytic leukemia (ALL), can be divided into E+ALL and E-ALL. We studied cell surface antigens on blasts from 12 children with untreated ALL: eight with E-ALL and four with E+ALL. Heterologous antisera were raised against thymus cells, E+ and E-ALL blasts, approximately absorbed and tested by immunofluorescence and a radiolabeled antibody assay with normal and leukemic lymphoid cells. By both methods, anti-thymus and anti-E+ALL sera reacted with human thymocytes. Specific binding of anti-E+ALL serum to T antigens was indicated by the fact that a single absorption with thymocytes abolished its binding to allogenic thymocytes, and the reactivity of anti-E+ALL serum with thymus, blood and bone marrow lymphocytes was similar to that of anti-thymus serum. After exhaustive absorption with blood leukocytes, anti-E+ALL and E-ALL sera were negative against normal lymphocytes and bone marrow cells from children with ALL in remission. Antithymus and anti-E+ALL sera reacted with blasts from patients with E+ALL, but not with E-ALL. In contrast, anti-E-ALL serum reacted with 40 to 96% of blasts from all children with E-ALL, whereas of the four patients with E+ALL, two were negative and two had the lowest percentage of immunofluorescent cells (10 to 22%). These results were confirmed with the radiolabeled antibody assay. Patients with active E-ALL had cells bearing E-ALL antigen(s) in the peripheral blood and bone marrow, but the number of immunofluorescent cells was lower in blood. Cells reactive with anti-E-ALL sera also were consistently found in sequential bone marrow or blood samples from children with E-ALL who had relapsed and were resistant to chemotherapy. Anti-E-ALL serum did not react with thymus cells, blood lymphocytes, remission bone marrow cells, Raji cells, PWM and PHA-induced blasts and CLL cells bearing mIg (uk). These data suggest that the antigen detected on E-ALL blasts by anti-E-ALL serum is neither a HLA-related nor a cell differentiation antigen. Thus, by using antiserum to E+ALL blasts, we have confirmed the presence of a T cell-specific antigen(s) on E+ALL cells. This antiserum did not recognize other leukemia-associated antigens common to E+ and E-ALL. We have also demonstrated an antigen(s) which is regularly expressed on E-ALL blasts and is either not detectable or is present in a lower proportion of E+ALL blasts.

E Receptors on Blasts from Untreated Acute Lymphocytic Leukemia (All): Comparison of Temperature Dependence of E Rosettes Formed by Normal and Leukemic Lymphoid Cells

The presence of receptors for sheep erythrocytes (E) and sufrace Ig on the bone marrow blasts was investigated in 29 children with untreated acute lymphocytic leukemia (ALL). In 6 of them more than 50% of the bone marrow blasts formed E rosettes, while in none of the 29 were surface Ig detected. The six children with rosette-forming blasts had WBS greater than 5 times 10-4/mm-3 at admission and 4 of them presented with thymic enlargement. E-positive ALL blasts and normal human thymocytes formed rosettes after incubation for 1 hr at 4 degrees C or 37 degrees C. In contrast, normal human peripheral T lymphocytes from blood, spleen, and pleural fluid also formed rosettes at 4 degrees C but these rosettes dissociated at 37 degrees C. In two patients with E-positive ALL, the disappearance from blood of cells forming rosettes at 37 degrees C during the 1st week of treatments paralleled the reduction in circulating blasts. Conversely, after 6 days of therapy almost one-half of the remaining cells formed rosettes at 4 degrees C and had the morphologic features of normal lymphocytes. We conclude that, in contrast to peripheral T cells, normal thymic cells and E-positive blasts share the property of forming E rosettes after 1-hr incubation at 37 degrees C. In patients with E-positive ALL this property may be used to evaluate drug effects upon leukemic and normal T lymphocytes.

Macroglobulinemia in a Child With Acute Leukemia

Abstract A 12-yr-old boy with acute leukemia was found to have paraproteinemia and Bence-Jones proteinuria. The paraprotein was characterized as immunoglobulin M, type κ and the Bence Jones protein as free κ-chains. Increased amounts of β2-microglobulin were found in the patient’s serum and urine. Electron microscopic and immunofluorescence studies provided evidence of the presence of lymphoid leukemia cells apparently responsible for the production of the paraprotein.