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.     

Paul-Bunnell antigen in lymphoma and leukemia spleens.

Lymphoid cells obtained from spleens of patients with lymphomas or leukemias were studied for the presence of heterophile (Paul-Bunnell (P-B)) antigen. A mixed agglutination (MA) test was established utilizing monolayers of cells attached to poly-L-lysine-coated wells of plastic U plates. After incubation of the monolayers with infectious mononeucleosis (IM) sera, indicator cells, sheep, or trypsinized bovine erythrocytes were added. The results were assessed according to sedimentation patterns of the indicator cells on the monolayers. Positive MA reactions were shown to be due to specific binding of P-B antibodies to the corresponding antigens on the spleen cells. Positive results were obtained with 15 of 37 spleens from patients with Hodgkin's disease, 5 of 8 lymphoma spleens, 4 of 15 chronic myelocytic leukemia spleens and 2 of 4 chronic lymphocytic leukemia spleens. Only 2 of 25 spleens from patients with various other diseases and 1 of 26 apparently normal thymus specimens gave positive results. This study confirmed demonstration of P-B antigen in lymphoma and leukemia by means of absorption experiments, which was reported previously.     

Detection of trisomy 12 by fluorescence in situ hybridization on archival cytopathologic material in chronic lymphocytic leukemia/small lymphocytic lymphoma

OBJECTIVE: To evaluate fluorescence in situ hybridization (FISH) for the detection of trisomy 12 in archival cytologic specimens of chronic lymphocytic leukemia/small lymphocytic lymphoma. STUDY DESIGN: The cytopathology database was searched for all cases of chronic lymphocytic leukemia/small lymphocytic lymphoma. Six cases of chronic lymphocytic leukemia/small lymphocytic lymphoma obtained by fine needle aspiration and one case of small lymphocytic lymphoma with plasmacytoid features were analyzed for trisomy 12 by FISH. These cases had been archived between 1 week to 16 months prior to analysis. RESULTS: We detected trisomy 12 in four of the six cases of small lymphocytic lymphoma/chronic lymphocytic leukemia. The case of small lymphocytic lymphoma with plasmacytoid features was negative for trisomy 12. CONCLUSION: Detection of trisomy 12 by FISH can be effectively performed on routinely prepared, stained and coverslipped archival cytologic material.     

Effect of treatment with thymustimulin (Tp-1) on T and B cells in lymphoproliferative disorders

The effect of the calf thymus extract thymustimulin (Tp-1) on lymphocyte subpopulations of 12 patients affected by lymphoproliferative disorders with low T-cell level was studied. T and B cells of four patients with non-Hodgkin lymphoma, three with Hodgkin's disease, one with myeloma and four with chronic lymphatic leukemia were evaluated before and after treatment for 3 months with Tp-1. The total number and the percentage of T-cells increased significantly (p less than 0.05) in patients with non Hodgkin lymphoma, Hodgkin's disease and myeloma and only numerically in patients with chronic lymphatic leukemia, while no significant change of the total WBC count and of the total number and percentage of B-cells occurred in any patients. These results suggest that Tp-1 treatment might be effective in restoring immunocompetence in patients with T-cell deficiency secondary to lymphoproliferative disorders.     

Growth arrest of BCR-ABL positive cells with a sequence-specific polyamide-chlorambucil conjugate

Chronic myeloid leukemia (CML) is characterized by the presence of a constitutively active Abl kinase, which is the product of a chimeric BCR-ABL gene, caused by the genetic translocation known as the Philadelphia chromosome. Imatinib, a selective inhibitor of the Bcr-Abl tyrosine kinase, has significantly improved the clinical outcome of patients with CML. However, subsets of patients lose their response to treatment through the emergence of imatinib-resistant cells, and imatinib treatment is less durable for patients with late stage CML. Although alternative Bcr-Abl tyrosine kinase inhibitors have been developed to overcome drug resistance, a cocktail therapy of different kinase inhibitors and additional chemotherapeutics may be needed for complete remission of CML in some cases. Chlorambucil has been used for treatment of B cell chronic lymphocytic leukemia, non-Hodgkin's and Hodgkin's disease. Here we report that a DNA sequence-specific pyrrole-imidazole polyamide-chlorambucil conjugate, 1R-Chl, causes growth arrest of cells harboring both unmutated BCR-ABL and three imatinib resistant strains. 1R-Chl also displays selective toxicities against activated lymphocytes and a high dose tolerance in a murine model.     

Lack of detectable somatic hypermutation in the V region of the Ig H chain gene of a human chronic B lymphocytic leukemia

Monoclonal human B cell tumors are a model system for the study of somatic hypermutation of the Ig genes of humans. It was previously shown that a number of B cell lymphomas exhibited striking V region point mutation, hypothesized to result from the somatic hypermutation mechanism. In this study we have extended the analysis to chronic lymphocytic leukemia. We have cloned and sequenced the productive Vh representing five different cells from a monoclonal chronic lymphocytic leukemia. All five Vh sequences were identical. Therefore, the Vh region in this leukemia was not the subject of detectable somatic mutation. These data suggest that chronic lymphocytic leukemia might lack the mechanism for somatic hypermutation and represent a stage of normal B lymphocyte differentiation in which the somatic hypermutation mechanism is not active.     

Cellules grumelées: old terminology revisited. Regarding the cytologic diagnosis of chronic lymphocytic leukemia and well-differentiated lymphocytic lymphoma in pleural effusions

Distinguishing chronic lymphocytic leukemia (CLL) or well-differentiated lymphocytic lymphoma (WDLL) from a benign chronic inflammatory process involving the serous cavities is often a difficult task for the cytopathologist faced with a lymphocyte-rich effusion fluid. Spriggs and Boddington decribed characteristic heavy chromatin clumping (cellules grumelées) of the lymphocytic nuclei in effusions as diagnostic of chronic lymphocytic leukemia. A study of 23 cases of lymphocyte-rich pleural effusions in our laboratory showed that, while this cytomorphologic feature is a function of cytopreparatory technique and fixation, it was observed only in cases of CLL or WDLL and not in benign inflammatory processes.     

Studies on delayed hypersensitivity of patients with Hodgkin's disease during remission.

The studies of skin sensitivity, the reactivity to intradermally transferred lymphocytes and lymphocytic transformation in vitro showed that delayed hypersensitivity had been slightly impaired in patients with Hodgkin's disease during long time remission. The regression of the disease favoured the recovery of the lymphocytic functions and the delayed type immune reactions associated with them. These data suggest that the enhancement of the delayed hypersensitivity of the tumour defence respectively could promote the treatment or prevention of the disease.     

The 1952 outbreak of encephalitis in California; vector control aspects

X-radiation remains the treatment of choice in most cases of leukemia and lymphoma, but new agents are playing an increasing role in therapy. Radioactive phosphorus does not produce radiation sickness and results with it are comparable to those of x-ray therapy in chronic leukemia. Urethane and nitrogen mustard may produce remissions in patients with chronic leukemia who have become resistant to radiation. Triethylene melamine may be administered orally with nitrogen mustard-like effects and is undergoing further trial. Aminopterin, ACTH and cortisone often cause short remissions in acute leukemia. Urethane is the best treatment available for multiple myeloma. Polycythemia vera is well controlled by radioactive phosphorus combined with venesection. Nitrogen mustard is often effective and triethylene melamine shows promise in Hodgkin's disease. Antianemic substances such as iron and liver extract are of no value in the treatment of anemia caused by leukemia, lymphoma and myeloma.     

The carcinolytic effect of actinomycin C; a report of clinical studies

Actinomycin C was administered to 35 patients with a variety of malignant states. There was no evidence of any toxic reaction. Of 11 patients with Hodgkins disease, four had brief objective remission ranging from two to four months. Individual patients with chronic myelogenous leukemia, multiple myeloma and neuroblastoma had short periods of clinical improvement. The usefulness of actinomycin C appears to be limited. It may best be used in instances of Hodgkin's disease where pancytopenia prevents the use of other agents.     

Biological weapons threat leads to inoculations for sailors

The authors describe a 51-year-old man with chronic lymphocytic leukemia who presented with respiratory distress and then died suddenly while in hospital. Autopsy revealed pulmonary leukostasis and a large intracardiac mass containing mostly mature lymphocytes and fibrin. Although leukostasis and lymphocyte thrombi have been described (albeit rarely) in chronic lymphocytic leukemia, an intracardiac "clot" has not. It seems plausible that this intracardiac mass caused the patient''s death.     

Osmotic fragility in subfractions of leucocytes in hematological diseases (author's transl)]

Leucocytes of normal persons and patients with acute and chronic granulocytic leukemia, chronic lymphocytic leukemia, and non-Hodgkin lymphoma were separated into subfractions by centrifugation in discontinuous Ficoll density gradient. Osmotic resistance was examined in hypotonic NaCl solutions with decreasing concentration and by determining LDH activity in the supernatant. Suspensions of myelocytes, polymorphnuclear granulocytes, and lymphocytes of normal persons and patients with chronic lymphocytic leukemia demonstrated the same osmotic resistance. Only myeloblasts were osmotically less fragile, and tumor cells of non-Hodgkin lymphoma more fragile.     

The Relation Between Lymphosarcoma and Leukemia

Of 283 cases of lymphocytic disease, 81% fell within three distinct categories: lymphocytic and lymphoblastic lymphosarcoma and lymphocytic leukemia. The remaining 19% showed transitions from more mature to less mature cell types or from local to general anatomic distribution. The clinical course was related to the cell type and the extent of disease rather than to the presence of blood stream invasion. Survival of patients with lymphocytic leukemia and of those with lymphocytic lymphosarcoma was the same, while that of patients with lymphoblastic lymphosarcoma was much shorter. Survival curves are simple exponentials and do not suggest two populations, one with disease less malignant than the other.     

Deoxyribonucleic Acid Polymerase Activity Associated with a Plasma Particulate Fraction from Patients with Chronic Lymphocytic Leukemia

Plasma from two cases of chronic lymphocytic leukemia have been examined for deoxyribonucleic acid (DNA) polymerase activity. In both cases, detectable enzyme activity was present. In the plasma from a patient known to have chronic lymphocytic leukemia for 10 years, the enzyme activity was sufficiently high to study product formation, buoyant density of the enzyme activity, deoxyribonucleoside triphosphate requirements, and kinetics of DNA synthesis. These studies are presented in this report.     

Amplification of RNA and DNA specific for erb B in unbalanced 1;7 chromosomal translocation associated with myelodysplastic syndrome

Previous work has established the presence of an unbalanced chromosome abnormality [+der(1),t(1;7)(p11;p11)] in some therapy-associated myelodysplastic disorders. Recently the EGF receptor has been found to reside at 7p11. Using a probe specific for erb B oncogene, which encodes a truncated form of the EGF receptor, we examined RNA and DNA derived from bone marrow and peripheral blood mononuclear cells from three patients with myelodysplastic syndromes (MDS) and one with acute lymphocytic leukemia (ALL), all bearing an abnormal clone in their bone marrow with a similar unbalanced 1;7 translocation. DNA-excess slot blot hybridization to 5'-32p-labeled cellular RNA revealed from ten- to thirtyfold enhancement in accumulation of mRNA specific for erb B in both peripheral blood and bone marrow cells of the three MDS patients when compared to normal controls. In addition, enhancement of H-ras mRNA accumulation was detected in some, though expression of other genes such as actin, N-ras, myc, src, B-lym, and 20 other genes was not found to be enhanced. Increased erb B expression was not apparent in mononuclear cells from patients with other hematologic disorders such as chronic lymphocytic leukemia, Hodgkin's disease, or lymphoma. Southern blot analysis of restriction-enzyme-cleaved DNA from three MDS patients with an unbalanced 1;7 translocation revealed that erb B gene was amplified at least twentyfold in peripheral blood white blood cells, while levels of actin hybridization were comparable to those of the controls. No such amplification was evident in the ALL patient. Our data suggest that +der(1),t(1;7)(p11;p11) chromosomal anomalies can be specifically associated with amplification of erb B DNA and RNA sequences.     

CD47 ligation induces caspase-independent cell death in chronic lymphocytic leukemia.

Thrombospondin forms a 'molecular bridge' between phagocytic and apoptotic cells through interaction with alphavbeta3/CD36. We report here that engagement of CD47, a newly described thrombospondin receptor, by immobilized monoclonal antibody against CD47 or by thrombospondin induced in all B-cell chronic lymphocytic leukemia clones the cytoplasmic features of apoptosis (cell shrinkage, decrease in mitochondrial transmembrane potential and phosphatidylserine externalization) without the nuclear features (chromatin condensation, appearance of single-stranded DNA, DNA fragmentation and cleavage of poly ADP-ribose polymerase). These cytoplasmic events of apoptosis were not prevented by the addition of caspase inhibitor z-VAD-fmk, or by the presence of survival factors (such as interleukin-4 and gamma interferon) or cell activation. Morphological studies confirmed the integrity of the nucleus and showed swelling of the mitochondria. This caspase-independent death pathway may be relevant to the development of alternate therapeutic strategies in chronic lymphocytic leukemia, which remains an incurable disease.     

Production and characterization of a monoclonal antibody that binds Reed-Sternberg cells

A murine monoclonal antibody, termed HeFi-1, was produced after immunization with the L428 Hodgkin's disease tissue culture cell line. HeFi-1 selectively stained only the Reed-Sternberg or Hodgkin's cells in 18 of 18 cases of Hodgkin's disease, including the nodular sclerosis, mixed cellularity, and lymphocyte-depleted histologic subtypes. HeFi-1 did not stain any cells in normal lung, brain, salivary gland, thyroid, gall bladder, pancreas, liver, testis, breast, endometrium, or kidney. Rare large cells at the edge of the lymphoid follicles were stained in normal tonsil, colon, and hyperplastic thymus. There was no staining of any cells in 14 cases of B cell non-Hodgkin's lymphoma; however, the malignant cells in three of 11 cases of non-Hodgkin's lymphoma which appeared to express T cell markers were also stained with HeFi-1. Tissue culture cell lines including the T cell acute lymphocytic leukemia lines MOLT4 and CEM, the histiocytic cell line U-937, and the amniotic cell line WISH were not stained. Seven Epstein Barr virus (EBV)-positive lymphoblastoid cell lines were stained with HeFi-1, but there was no staining of three EBV+ African Burkitt's lymphoma cell lines or three EBV- American Burkitt's cell lines. HeFi-1 did not block the ability of the L428 cells to stimulate a mixed lymphocyte reaction or function as accessory cells for mitogen-induced human T cell proliferative responses. Modulation of the HeFi-1 cell surface antigen on the L428 cells was not observed. HeFi-1 specifically immunoprecipitated a cell surface protein of approximately 120,000 daltons from both the L428 and EBV+ lymphoblastoid cell lines. HeFi-1 monoclonal antibody should prove useful not only in the diagnosis, staging, and potential therapy of Hodgkin's disease, but also for determining the cell of origin of the Reed-Sternberg cell.     

Risks of leukemia in Japanese atomic bomb survivors, in women treated for cervical cancer, and in patients treated for ankylosing spondylitis.

The dose-response relationship for radiation-induced leukemia was examined in a pooled analysis of three exposed populations: Japanese atomic bomb survivors, women treated for cervical cancer, and patients irradiated for ankylosing spondylitis. A total of 383 leukemias were observed among 283,139 study subjects. Considering all leukemias apart from chronic lymphocytic leukemia, the optimal relative risk model had a dose response with a purely quadratic term representing induction and an exponential term consistent with cell sterilization at high doses; the addition of a linear induction term did not improve the fit of the model. The relative risk decreased with increasing time since exposure and increasing attained age, and there were significant (P < 0.00001) differences in the parameters of the model between datasets. These differences were related in part to the significant differences (P = 0.003) between the models fitted to the three main radiogenic leukemia subtypes (acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia). When the three datasets were considered together but the analysis was repeated separately for the three leukemia subtypes, for each subtype the optimal model included quadratic and exponential terms in dose. For acute myeloid leukemia and chronic myeloid leukemia, there were reductions of relative risk with increasing time after exposure, whereas for acute lymphocytic leukemia the relative risk decreased with increasing attained age. For each leukemia subtype considered separately, there was no indication of a difference between the studies in the relative risk and its distribution as a function of dose, age and time (P > 0.10 for all three subtypes). The nonsignificant indications of differences between the three datasets when leukemia subtypes were considered separately may be explained by random variation, although a contribution from differences in exposure dose-rate regimens, inhomogeneous dose distribution within the bone marrow, inadequate adjustment forcell sterilization effects, or errors in dosimetry could have played a role.     

Alpha interferon in T helper phenotype chronic lymphocytic leukemia: a report of three cases

Three patients affected by T helper chronic lymphocytic leukemia were treated with low dose interferon alpha-2b (3 MU/m2 3 times weekly). The disease presented different pathologic expressions with diffuse skin lesions in one patient, a mild clinical course and a prolymphocytic variant with aggressive features, respectively, in the other two cases. A consistent response was observed within 3-6 weeks; by that time a reduction of blood and marrow lymphocytosis in the three patients and a regression of the cutaneous lesions were documented. Therefore, it should be emphasized that the use of alpha IFN, whose effectiveness on cutaneous T cell lymphomas has been already demonstrated, may represent an active agent in the treatment of leukemic T helper phenotype chronic lymphocytic proliferations.     

Detection of a soluble form of the receptor for interleukin 2 in the serum of patients with hairy cell leukaemia

The sera of patients with hairy cell leukaemia (HCL) contain a factor which inhibits the binding of anti-IL-2R antibody to its target (activated T lymphocytes). The presence of this factor, which probably corresponds to the soluble form of IL-2R (sIL-2R), can be easily detected using a simple immunocytochemical inhibition assay. In a series of patients with chronic lymphoproliferative diseases the presence of sIL-2R appeared to be specific for HCL, using the sensitivity of our test, since it could not be detected in sera from normal subjects and patients with B-cell lymphocytic leukaemia or Hodgkin's disease. Thus it might be used as an additional tool for characterizing HCL patients.