Cap formation on lymphocytes from patients with leukemic diseases induced by four different lectins.

When ficoll purified peripheral blood lymphocytes were treated with fluorescein conjugated lectins from lentils (LCH), castor beans (RCA) and phaseolus coccineus beans (L-and E-PHA) for 15 min and the percentages of the cap forming cells were examined, the values of leukemic lymphocytes were reduced compared to the values obtained with normal lymphocytes. The reduction was more than half in patients with acute and chronic myelogenous leukemia and immunoblastoma, it was only one quarter in patients with chronic lymphocytic leukemia, Hodgkin's disease and lymphosarcoma. The lowest number of cap forming cells was found in lymphoblasts of established lymphoblastoid cell lines. The four different lectins showed nearly the same capacity in the induction of caps. After successive binding, the different lectins showed cocapping on the lymphocyte surface.     

Lymphosarcoma with lymphoblastic leukemia in a New Zealand white rabbit.

An 8 to 10-week-old female New Zealand white rabbit, Oryctolagus cuniculus, which exhibited clinical signs of anorexia, depression, and torticollis was found to have lymphosarcoma with lymphoblastic leukemia. The multiple visceral involvement with neoplastic lymphoid cells observed in this animal was similar to previously reprted cases of lymphosarcoma in the rabbit. An unusual finding was the occurrence of lymphoblastic leukemia since lymphosarcoma in the rabbit has previously been reported as aleukemic.     

Stimulant of antibody producers isolated from the supernatants of cell cultures of normal human bone marrow and in various diseases

The biological activity of a stimulant of antibody producers (SAP) isolated from normal human bone marrow was studied and compared with that of a stimulant of antibody producers from the bone marrow of patients with acute myeloblastic leukemia, acute lymphoblastic leukemia, lymphosarcoma, and multiple myeloma. The activity of the SAP from human bone marrow in health was similar to that of analogous transmitters from the bone marrow of other species of the mammals and birds. The activity of the SAP in patients with multiple myeloma was elevated, whereas in patients with acute myeloblastic leukemia, it was lowered.     

Prevalence of antigen receptor variants in human T cell lines and tumors

Previously, we have shown that a human T acute lymphoblastic leukemia cell line, HPB-ALL, exhibits clonal heterogeneity within its Ag receptor, as revealed by varying reactivity patterns with a panel of anti-idiotype mAb. We now extend these findings to another human T acute lymphoblastic leukemia cell line, SUP-T13, and to two fresh human chronic lymphocytic leukemias, JE and EF. In the two cell lines, two types of Ag receptor variants could be found: those that retained a receptor molecule but lost reactivity with an anti-idiotype mAb (idiotype variants), and those which had lost surface receptor expression completely (receptor-negative variants). The idiotype variants, at least in HPB-ALL, have differentially glycosylated receptor alpha-chains from the parent. The receptor-negative cells, in HPB-ALL as well as in SUP-T13, produce cytoplasmic receptor and CD3 proteins but do not transport them to the surface. Neither idiotype nor receptor-negative variants could be detected in either of the fresh tumors of chronic lymphocytic leukemias. The limit of sensitivity in these analyses was about 0.05%. We conclude that antigen receptor variants can spontaneously occur in cell lines derived from acute lymphoblastic leukemias, but are infrequent in chronic lymphocytic leukemias in vivo, and that therapy with anti-idiotype mAb may be a viable strategy for these malignancies.     

Antileukemic Activity of Sulforaphane in Primary Blasts from Patients Affected by Myelo- and Lympho-Proliferative Disorders and in Hypoxic Conditions

Sulforaphane is a dietary isothiocyanate found in cruciferous vegetables showing antileukemic activity. With the purpose of extending the potential clinical impact of sulforaphane in the oncological field, we investigated the antileukemic effect of sulforaphane on blasts from patients affected by different types of leukemia and, taking into account the intrinsically hypoxic nature of bone marrow, on a leukemia cell line (REH) maintained in hypoxic conditions. In particular, we tested sulforaphane on patients with chronic lymphocytic leukemia, acute myeloid leukemia, T-cell acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia, and blastic NK cell leukemia. Sulforaphane caused a dose-dependent induction of apoptosis in blasts from patients diagnosed with acute lymphoblastic or myeloid leukemia. Moreover, it was able to cause apoptosis and to inhibit proliferation in hypoxic conditions on REH cells. As to its cytotoxic mechanism, we found that sulforaphane creates an oxidative cellular environment that induces DNA damage and Bax and p53 gene activation, which in turn helps trigger apoptosis. On the whole, our results raise hopes that sulforaphane might set the stage for a novel therapeutic principle complementing our growing armature against malignancies and advocate the exploration of sulforaphane in a broader population of leukemic patients.     

Divergen properties of two human lymphocytic cell lines isolated from a single specimen of peripheral blood

Summary Two lymphocytic cell lines were obtained from a single specimen of peripheral blood buffy coat from a patient with acute lymphoblastic leukemia who concurrently developed classical infectious mononucleosis. These two cell lines exhibit distinguishing differences in culture in parallel with reviously described differences in the cells when implanted into eonatal Syrian hamsters. These observations suggest that the cell lines were derived from two pre-existing different classes or “stelines” of cells—one of which resembles other cell lines derived from patients with acute lymphoblastic leukemia, whereas the other resembles cell lines derived from other, patients with infecttious mononucleosis. These studies were suported in part by research Grants C-6516 from the National Cancer Institute and FR-05526 from the Division of Research Facilities and Resources, National Institutes of Health.     

中药联合化疗对ALL患者CD4、CD25细胞调节及sE-cad的影响

目的:研究中药联合化疗方案对急性淋巴细胞白血病(Acute lymphocytic leukemia,ALL)患者CD4+、CD25+细胞调节的影响、血清s E-cad的影响以及临床意义。方法:选取我院血液科收治的急性淋巴细胞白血病患者60例,随机分为两组,其中对照组30例给予常规化疗方案治疗,中药组30例在常规化疗方案的基础上加用中药辅助治疗。对比治疗前后患者血常规、CD4+、CD25+细胞及血清s E-cad的改变。结果:1治疗后两组患者CD4+、CD25+T细胞较治疗前均显著下降,差异有统计学意义(P0.05),与对照组相比,中药组CD4+、CD25+T细胞下降明显,差异有统计学意义(P0.05);2治疗后两组患者血清s E-cad均改善,且中药组(55.58±10.47)较对照组(67.27±11.32)明显下降,差异有统计学意义(P0.05);3两组有效率比较重,中药组总有效率(86.67%)明显优于对照组(66.67%),差异有统计学意义(P0.05)。结论:中药联合化疗治疗急性淋巴细胞白血病CD4+、CD25+T细胞调节及血清s E-cad的改变明显,对临床具有指导意义。     

Phosphatidylethanolamine methylase and cyclic nucleotide phosphodiesterase activities in human B lymphoid hemopathies

Phospholipid methylase and cyclic nucleotide phosphodiesterase activities were studied in human B lympho?d hemopathies (51 patients: acute lymphoblastic leukemia, B lymphoma, chronic lymphocytic leukemia, hairy cell leukemia) and compared with activities in lymphoblast?d and Burkitt lymphoma cell lines and with normal B lymphocytes: methylase activity proved to be lower in ALL and high grade lymphoma and inversely related to the percent of cells in S phase state; the A/G ratio of phosphodiesterases was low in ALL and CLL and high in hairy cell leukemia and it was related to the percent of cells in S phase state.     

In vitro culture studies of granulocyte/macrophage and erythroid progenitor cells in lymphoproliferative disorders

Granulocyte/macrophage progenitor cells (CFU-GM) and erythroid progenitor cells (BFU-E) have been assayed in peripheral blood (PB) and/or bone marrow (BM) from 12 patients with acute lymphocytic leukemia (ALL), 16 patients with chronic lymphocytic leukemia (CLL) and 31 patients with various forms of non-Hodgkin lymphoma (NHL) without BM involvement. Progenitor cell growth in PB and BM from the NHL patients did not differ statistically from controls (p greater than 0.1). CFU-GM and BFU-E per ml PB were markedly increased in ALL and CLL patients (p less than 0.001) while CFU-GM and BFU-E per plated BM cells from these patients were severely depressed (p less than 0.001). Lymphoblasts from one ALL patient failed to inhibit CFU-GM and BFU-E-derived colony growth from control PB mononuclear cells. The high levels of circulating progenitor cells in ALL and CLL patients clearly distinguish them from other cytopenic hematological malignancies, in which decreased progenitor cell levels have been demonstrated previously (acute myeloid leukemia, hairy cell leukemia). The cause of this finding and its pathophysiological implication still remains to be established.     

A novel human T cell antigen preferentially expressed on mature T cells and shared by both well and poorly differentiated B cell leukemias and lymphomas

A new lymphocyte differentiation antigen shared by all normal T cells and some malignant B cells was defined by a monoclonal antibody designated 12.1. This antibody reacted with all peripheral blood T cells but not with normal B cells and B cell lines. Analysis with a fluorescence activated cell sorter showed that the expression of 12.1 antigen changes during T cell maturation. Most thymocytes, blasts of acute T cell leukemia, and cells from established leukemic T cell lines bear a small amount of 12.1 antigen. In contrast the majority of peripheral blood T cells, activated T cells, and leukemic T cells of the Sezary syndrome bear a large amount of 12.1 antigen. Unexpectedly, antibody 12.1 reacted with leukemic cells from most patients with B-type chronic lymphocytic leukemia (CLL) and some patients with lymphosarcoma cell leukemia (LSCL). Among these leukemias, expression of the 12.1 antigen was not correlated with the stage of B cell maturation, with the amount of surface immunoglobulin on the cells, or with the presence or absence of monoclonal gammapathy. In a comparative serologic analysis the antigen defined by antibody 12.1 was distinct from the p67 T cell antigen (defined by monoclonal antibody 10.2) that is also known to be expressed by B-type CLL cells.     

Cyclic AMP phosphodiesterase in human lymphocytes and lymphoblasts.

Cyclic nucleotide phosphodiesterase activities were examined in lymphocytes from 12 transformed human B cell lines, two T cell lines, six patients with lymphocytic leukemia, and 10 normal donors. A consistent difference bwtween cells from the normal and leukemic state was observed. The cyclic AMP phosphodiesterase activity from normal lymphocytes is inhibited greater than 80% by muM cyclic GMP while this concentration of nucleotide has little or no effect on the enzyme from transformed lymphocytic cell lines or from lymphocytic cells of leukemia patients. The reported lack of cyclic GMP phosphodiesterase in human lymphocytes from several sources is confirmed. The apparent absence of a cyclic GMP degradation mechanism and of cyclic GMP control of cyclic AMP hydrolysis may be related to defective lymphocyte growth control.     

Deoxycytidylate deaminase activity in non-stimulated and phytohemagglutinin-stimulated human lymphocytes,and in leukemic cells

Deoxycytidylate deaminase isolated from normal human lymphocytes and from mononuclear leucocytes from patients with acute lymphoblastic leukemia, chronic lymphocytic leukemia and acute monocytic leukemia has been characterized in regard to the substrate, dAMP and the allosteric regulators dCTP and dTTP. The enzymes exhibited sigmoidal initial velocity versus dCMP concentration whereas in the presence of the activator, dCTP, Michaelis-Menten kinetics were obtained.At saturating substrate concentrations dTTP acted as an allosteric inhibitor of the enzyme isolated from non-stimulated as well as from stimulated lymphocytes. However, the enzymes isolated from the leukemic cells had lost the allosteric regulation by dTTP.At low substrate concentrations the competitive inhibitor, dAMP, activated all the enzymes. This activation was abolished in the presence of dCTP which indicates that dAMP might be involved in the regulation of dCMP deaminase activity and thus influence the dCTP and dTTP pools under physiological conditions.Abbreviations dCMP deaminase deoxycytidylate deaminase - PHA Phytohemagglutinin - ALL acute lymphoblastic leukemia - CLL chronic lymphocytic leukemia - AMOL acute monocytic leukemia - WBC white blood cells     

Vincristine in the Treatment of Neoplastic Disease

Seventy-eight patients with advanced malignant disease were treated with vincristine, an alkaloid derived from Vinca rosea Linn. Fifty-nine of these survived one month from the beginning of treatment and could be evaluated. Twenty made a good response, with return to activity and more than 75% regression of tumour deposits. Seven made a fair response with resumption of partial activity and at least a 25% regression of tumour deposits. Favourable results were seen in patients with Hodgkin''s disease, reticulum cell sarcoma, lymphosarcoma, carcinoma of the breast, acute leukemia and choriocarcinoma. The duration of remission was usually brief but in five instances exceeded six months. Toxic reactions include a high incidence of alopecia and neurologic complications. The bone marrow depression is constant and predictable.     

Morphometric study of bone marrow in chronic lymphocytic leukemia

A morphometric study utilizing the point counting method was carried out on bone marrow biopsies of 44 patients with chronic lymphocytic leukemia (CLL) in order to correlate the objective volume of the lymphocytic infiltration with the histologic patterns (nodular, interstitial, mixed and diffuse) and the clinical stages (A, B and C). The prognostic significance of isolated lymphocytic tumor cell burden was also analyzed. The results suggest that there is a significant correlation between the amount of tumoral lymphoid tissue (VL greater than 60% versus VL less than 60%) and the interstitial and diffuse histologic patterns, as well as with the clinical stages A and C. However, the lymphoid burden did not correlate with the nodular and mixed patterns, nor with the clinical stage B. When patients with VL greater than 60% were compared with those with VL less than 60%, the difference in cumulative survival rates was not significant after the fourth year.     

Quantification of the leukocyte common antigen (CD45) in mature B-cell malignancies.

CD45 is a glycoprotein expressed on all lymphohematopoietic cells. Its expression increases during normal B-cell differentiation and remains stable on mature cells. Although it is widely known that CD45 antigen expression is decreased in B-acute lymphocytic leukemia (ALL), only scarce and contradictory information is available on CD45 expression on mature B-cell malignancies. In healthy adults (n = 15), CD45 expression on B lymphocytes was lower than that on T cells. In patients with chronic lymphocytic leukemia (CLL; n = 22), CD45 expression on malignant cells was lower than that on the whole lymphocyte population of healthy adults (n = 28) and on normal B lymphocytes (n = 15). In 6 of the 22 CLL patients, the malignant cell population could be separated from the normal lymphocyte population on the CD45-side scatter (SSC) plot. In 16 CLL patients, there was some degree of overlap between the malignant and normal cells with respect to CD45 expression. For these patients, there was an inverse correlation between CD45 expression on the whole lymphocyte population and the percentage of malignant cells in this population. In two patients with mantle cell lymphoma (MCL), CD45 expression on the malignant cells appeared lower than that on normal B cells and on the whole lymphocyte population. In six patients with hairy cell leukemia (HCL), CD45 expression on hairy cells was comparable to that on the whole lymphocyte population of healthy adults, but slightly higher than that of normal B cells. Evaluation of CD45 expression may help to characterize mature B-cell malignancies.     

Plasma fibronectin in acute leukemia. Effects of the cytostatic therapy on plasma fibronectin level

Twice a week plasma (Pl.)-fibronectin was determined quantitatively in the course of disease with immunoelectrophoresis according to Laurell in 12 patients suffered from acute non-lymphoblastic leukemia (ANLL) and in 12 patients affected with acute lymphoblastic leukemia (ALL). At diagnosis Pl.-fibronectin concentration was found to be significantly lowered only in those patients affected with ANLL. During the induction therapy Pl.-Fibronectin could be observed to decline significantly in all patients: in acute non-lymphoblastic leukemia from mean 270 micrograms/ml, s 93 micrograms/ml, to mean 185 micrograms/ml, s 89 micrograms/ml (p less than 0.01), and in acute lymphoblastic leukemia from mean 290 micrograms/ml, s 98 micrograms/ml, to mean 180 micrograms/ml, s 94 micrograms/ml (p less than 0.01). After administering L-asparaginase there is a strong decline of Pl.-fibronectin. Pl.-fibronectin concentration could be observed to be significantly lower in patients without remission in comparison to those with remission. A correlation between Pl.-fibronectin concentration and tumour mass could not be identified.     

B-cell lymphoma lacking Fc- and C3d-receptors.

Various cell surface markers were studied in a patient with lymphosarcoma cell leukemia. The B-cell derived feature of the neoplastic cells could be identified by demonstration of monoclonal surface immunoglobulin of IgM-kappa type of high density synthesized by the cells. Interestingly, there were no Fc- nor C3d-receptors demonstrable using various techniques. Only 22% of the leukemic cells expressed C3b receptors. The failure of rosette formation with mouse erythrocytes was an additional surface feature distinguishing from ordinary chronic lymphatic leukemia of the B cell type. The phenotype of the leukemia cells is discussed as corresponding to that of a less differentiated B lymphocyte.     

Suspension cultures of human malignant lymphoma in the leukocyte migration test

The effects of supernatants of primary and secondary malignant human lymphoma cell cultures were analyzed as parameters of spontaneous secretion of factors by these cells using the leukocyte migration test (LMT). Spontaneous cultivation for up to five weeks was successful in four cases. The postulated production of mediators, i.e. the inhibitory and stimulating effects on leukocyte migration were characterized by testing the influence of (a) concentration, (b) temperature and (c) absorption with normal blood leukocytes on the effect. Reproducible stimulatory and inhibitory effects on the migration of normal leukocytes were dependent on concentration and temperature and were apparently mediated by one or more factors. The supernatants of a lymphoblastic lymphosarcoma of the T-cell type and of a lymphoblastic lymphosarcoma clearly revealed congruous and reproducible inhibitory effects. A further case of lymphoblastic lymphosarcoma that could not exactly be defined with immunological methods either and a case of centroblastic/centrocytic lymphosarcoma exhibited stimulating effects which could be reduced in a time-dependent manner through preincubation with blood leukocytes. The results of these studies support the assumption that malignant lymphoma cells are capable not only of secreting immunoglobulin, but also of other biologically effective secretion. The effects of such secretion are differentiated into stimulating and inhibitory ones. They might be important for the spreading of a tumor or for resistance of the organism to the disease.