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Shedding and reappearance of Fc, C3 and SRBC receptors on peripheral lymphocytes from normal donors and chronic lymphatic leukaemia (CLL) patients.

The shedding and resynthesis of Fc, C3 receptors, and receptors binding SRBC has been studied in normal peripheral lymphocytes, in lymphocytes from CLL patients and in a Raji cell line during 4 h incubation at 37 degrees. During the first part of the incubation, an increased release of receptors was observed. The shed, functionally active, receptors were detectable in the incubating medium on the basis of their ability to inhibit rosette formation specifically. After 4 h incubation the original ratios of the different rosette forming cells were observed, showing that the cells were capable of resynthesizing their shed receptors. This later phenomenon may be the result of an active metabolic process. The shedding was not detectable on Raji cells. Different patterns of receptor shedding and resynthesis were demonstrated by the lymphocytes of CLL patients. The correlations between the reappearance of the receptors on the cells, and the metabolism of the lymphocytes, is discussed.

Antigens↗

Enumeration of TG lymphocytes in B-chronic lymphocytic leukaemia (B-CLL) by a rapid double-rosetting technique.

T lymphocytes bearing the Fc receptors for IgG (E+ F c gamma + or TG cells) in 38 patients with B-cell chronic lymphocytic leukaemia (B-CLL) were quantified by a simple one-step double-rosette technique. This assay detects the TG lymphocytes from total peripheral blood mononuclear cells (MNC) on the basis of their capacity to bind simultaneously sheep red cells and chicken erythrocytes sensitized with anti-CRBC rabbit IgG. The data obtained were comparable to those evaluated by the more elaborate conventional two-step method, which requires initial T-cell purification. The double-rosette technique is specific for T cells and it may be adapted as a routine clinical laboratory assay for TG cells in B-CLL patients who have very low T/B cell ratio owing to clonal expansion of B cells.

Aged↗

T cell subpopulations in CLL: methods of T cell enrichment artificially alter proportions of OKT4 and OKT8 positive cells.

There is growing speculation about the meaning of reported imbalances in subpopulations of T lymphocytes in patients with chronic lymphocytic leukaemia (CLL). This study compared two techniques for producing T cell enriched subpopulations from both patients with CLL and normal individuals and the effects of these techniques on relative proportions of OKT4 and OKT8 positive cells. A sheep red cell rosetting technique resulted in significantly larger OKT8 and smaller OKT4 positive populations than did nylon wool column elution. Similar results were obtained in normal individuals. The nylon wool column elution technique produced less distortion of unfractionated OKT4/OKT8 ratios in normals than did the rosetting technique. Studies of T lymphocyte subpopulations should be interpreted with great caution as the methods used to study them can influence the results.

Antibodies, Monoclonal↗

[Present status of chemotherapy in ALL and CLL].

Present status of chemotherapy for acute lymphocytic leukemia (ALL) and chronic lymphocytic leukemia (CLL) was reviewed. In ALL, a 2-drug combination consisting of vincristine and prednisone (VP) obtained an overall complete remission rate of approximately 50% and when daunomycin or adriamycin was added to the regimen, the overall remission rate escalated to approximately 70%. Furthermore, L-asparaginase appeared to have a similar activity to the anthracyclines in combination with VP. Thus, VP plus one of these agents seemed to be a recommendable inductive chemotherapy against ALL. In CLL, remission rates obtained by the use of chlorambucil alone were ranged from 45 to 69%, while a 2-drug combination consisting of chlorambucil and prednisone obtained remission rates ranging 38 to 80%. Thus, the combination appeared to have superiority over chlorambucil alone. When this first line chemotherapy fails to obtain remissions, a 3-drug combination consisting of cyclophosphamide, vincristine and prednisone is worthwhile to try as a second line treatment.

Adult↗

Monoclonal immunoglobulin of CLL lymphocytes detected by radioimmunofixation with 125I-protein A of Triton X-100 solubilized lymphocyte material.

A sensitive radioimmunofixation method (detection limit 1 mg/1 for monoclonal IgM) utilizing 125I Protein A and immunofixation for demonstration of monoclonal immunoglobulin in Triton X-100 extracts of isolated chronic lymphocytic leukemia (CLL) lymphocytes is presented. Thirty-two untreated patients with clinically typical CLL were studied and the results compared to those obtained by immunofluorescence. Monoclonal IgM was detected with similar frequency by the two methods (69% and 75%), but IgD was detected less frequently by radioimmunofixation (25% and 40%). When the results obtained with the two methods were combined, all of the cases were immunoglobulin positive. Two complete IgM type M-components were demonstrated in two cases, an excess of seemingly free heavy chains was found in four cases, and an excess of seemingly free light chains in six cases.

Antibodies, Monoclonal↗

Activation of lymphocytes in CLL by protein A from Staphylococcus aureus.

The mitogenic activity of protein A (PA), leucoagglutinin (LA) and pokeweed mitogen (PWM) was tested in six cases of chronic lymphatic leukaemia (CLL), and the response to PA and LA in six healthy controls. The effect of foetal calf serum (FCS) and pooled human AB serum in the cultures was tested in two patients. The response to LA in the patients was about one tenth of that in the controls. The response to PA was strong both in patients and in healthy controls. However, in lymphocytes from CLL patients this response was serum-dependent, being very strong with FCS in the culture medium, but absent when the medium contained human AB serum. Lymphocytes from normal controls responded to PA in the presence of AB serum.

Agglutinins↗

Fludarabine phosphate in chronic lymphoproliferative diseases. The French Group on CLL.

The review of Phase II trials in patients treated by Fludarabine phosphate (Fludara I.V) has been demonstrated to have unequivocal benefit for some patients with Chronic Lymphocytic Leukemia (CLL) non Hodgkin lymphoma and macroglobulinemia. Comparison of Fludara I.V with other treatment regimens is now studied in randomised protocols for B and C stages of CLL.

Antimetabolites, Antineoplastic↗

Oat cell carcinoma of the lung in patients with CLL.

Two malignancies in one patient was a remote possibility in the distant past, but currently more of these cases are being seen. At our institution, several patients with Chronic Lymphocytic Leukemia (CLL) were seen to develop oat cell carcinoma of the lung. The decreased immunity and B-cell dysfunction in CLL probably accounts for this secondary malignancy. We are presenting these cases to alert physicians of combined malignancies in a patient with increased aggressiveness of oat cell carcinoma.

Aged↗

[CD5-negative B-cell chronic lymphocytic leukemia (B-CLL) associated with massive splenomegaly and without lymphadenopathy].

A 39-year-old male was referred to our hospital in June, 1993, because of leukocytosis. Physical examinations showed massive splenomegaly without any lymphadenopathy. The white blood cell count was 13,800/microliters with 87% morphologically mature lymphocytes. Bone marrow aspirate revealed hypercellularity with 67% lymphocytes morphologically similar to peripheral lymphocytes. The lymphocytes displayed monoclonal rearrangements of immunoglobulin genes and the phenotype of CD5-CD19+CD20+ CD21+ and Smlg+. Splenectomy was effective against neutropenia and thromboytopenia. The clinical and laboratory findings of this case were unusual compared to those of typical B-CLL in massive splenomegaly, no lymphadenopathy and CD5-phenotype, suggesting the heterogeneity of B-CLL.

Adult↗

Long-term follow-up of patients with chronic lymphocytic leukemia (CLL) receiving fludarabine regimens as initial therapy.

One hundred seventy-four patients with progressive or advanced chronic lymphocytic leukemia (CLL) have received initial therapy with fludarabine as a single agent or fludarabine combined with prednisone. The overall response rate was 78% and the median survival was 63 months. No difference in response rate or survival was noted in the 71 patients receiving fludarabine as a single agent compared with the 103 patients who received prednisone in addition. The median time to progression of responders was 31 months and the overall median survival was 74 months. Patients over the age of 70 years had shorter survivals. Patients with advanced stage disease (Rai III and IV) had a somewhat shorter survival than earlier stage patients. More than half the patients who relapsed after fludarabine therapy responded to salvage treatment, usually with fludarabine-based regimens. Second remissions were more common in patients who had achieved a complete remission on their initial treatment. The CD4 and CD8 T-lymphocyte subpopulations decreased to levels in the range of 150 to 200/microL after the first 3 courses of treatment. Although recovery towards normal levels was slow, the incidence of infections was low in patients in remission (1 episode of infection for every 3.33 patient years at risk) and decreased with time off treatment. There was no association of infections or febrile episodes with the use of corticosteroids or the CD4 count at the end of treatment and a poor correlation with the increase in CD4 counts during remission. Infectious episodes were less common in patients who had a complete response compared with partial responders. Richter's transformation occurred in 9 patients and Hodgkin's disease occurred in 4 patients. Five other patients died from other second malignancies. Fludarabine appears to be an effective initial induction therapy with a reasonable safety profile for patients with CLL.

Adult↗

Prolonged survival in Richter syndrome with subsequent reemergence of CLL: a case report including serial cell-surface phenotypic analysis.

The development of a large cell lymphoma in a patient with chronic lymphocytic leukemia (Richter syndrome) is associated with a poor prognosis. There is dispute regarding the clonal origin of the large cell component. We described a patient in whom prolonged remission of the large cell component was achieved with combination chemotherapy, followed by subsequent reemergence of the less aggressive CLL clone. Serial cell surface phenotypic data are presented suggesting origin of both histologies from a common B-cell clone.

B-Lymphocytes↗

TNF-alpha induces spreading of B-CLL via the CD11c/CD18 molecule.

Activation of malignant B cells can lead to extensive morphological changes of these cells. A combination of PMA and TNF-alpha can induce adherence of purified B-CLL cells, which acquire a dendritic cell-like appearance. This phenomenon that we have termed spreading, is accompanied by upregulation of expression of both beta 1 and beta 2 integrin molecules. Spreading was inhibited by the addition of antibodies against CD18 or CD11c. In other B cell malignancies (HCL and NHL), morphological changes could be induced by PMA in the absence of TNF-alpha. Culturing in the presence of prednisolone resulted in an inhibition of spreading, most likely mediated via a down regulation of CD18 and CD11c expression. These data indicate that the CD11c/CD18 complex might be important for the adhesive properties of B cells.

Antigens, CD↗

B-CLL cell surface markers and mitogen-induced thymidine uptake: a comparison between lymph node cells and peripheral blood lymphocytes in 25 patients.

Lymphocytes from peripheral blood and lymph nodes from 25 patients with B-CLL were analysed by immunofluorescent staining of surface membrane immunoglobulin (SmIg), the B-cell marker B1, and the T-cell markers Leu 1/2/3/4. Tritium-thymidine uptake was measured in mitogen-stimulated 4-day cultures. Differences in these parameters between cells from the two sources in each patient were calculated with the paired T-test. All cell samples showed a clonal B-cell population. More blood lymphocytes than lymph node cells expressed the monoclonal SmIg (mu or gamma, p = 0.04; kappa or lambda, p less than 0.01), and T-cells were more frequent in lymph nodes (p = 0.01), where the T helper/suppressor ratio was higher. Furthermore, lymph node cells showed a higher thymidine uptake in response to the mitogens LPS, PWM, and Cowan (p = 0.01, p = 0.01, and p = 0.004, respectively), but there was no difference in the responses to EBV, DxS, TPA and PHA (p greater than 0.5). The higher response in lymph node cells to some mitogens might in part be explained by differences in the numbers of accessory cells, such as T-helper cells, but also by the existence of leukaemic B-cell subsets with different mitogen response patterns and different distributions within the lymphoid compartments. The characterization of subsets within a malignant cell clone might be of clinical importance.

Adult↗

Surface immunoglobulin pattern of the leukaemic cell population in chronic lymphocytic leukaemia (CLL) in relation to disease activity.

The surface membrane immunoglobulin (smIg) isotype pattern of the leukemic lymphocytes was studied in 66 unselected patients with CLL. Five distinct patient groups were identified according to the dominant heavy chain isotype(s); I: smmu + (n = 22), II: smmu +/smdelta + (n = 25), III: smdelta + (n = 4), IV: smgamma + (n = 5), V: no detectable heavy chains (n = 10). The majority of group I patients had a progressive disease at test while all patients in group II were in a non-progressive state. Moreover, the smIg pattern changed with the clinical activity of the disease: when the disease progressed, the relative number of smmu + cells increased and when patients entered an indolent stage after treatment the smmu + cell population decreased. In patients with stationary disease the smIg pattern remained essentially unchanged or the relative number of smdelta + cells increased. These observations might suggest that the smIg isotype pattern of the leukemic cell population reflects the biological behaviour and the clinical activity of the disease.

Adult↗

A pilot study of epirubicin and chlorambucil in the treatment of chronic lymphocytic leukemia (CLL).

In a pilot study of the combination of epirubicin and chlorambucil in the treatment of chronic lymphocytic leukemia (CLL), 10 patients with advanced or progressive disease were treated in four centres. Up to a total of 15 courses in individual patients were given. Toxicity was relatively mild with nausea due to epirubicin being the most commonly reported side-effect. One patient developed a probable septicemia during a period of neutropenia. In another patient who died from probable Pneumocystis carinii pneumonia (PCP) after only one course of therapy, death could not be regarded as necessarily treatment-related. In seven previously untreated patients, one complete response and four partial remissions were seen. All three previously treated patients showed partial remissions.

Adult↗

Chromosomal aberrations in a case of T-cell CLL with concomitant IgA myeloma.

Four consistent markers, involving chromosomes 8, 11, 14, 18, 21 and 22, were found in the majority of metaphases obtained after stimulation of peripheral blood and bone marrow from a patient with a T-cell chronic lymphocytic leukemia (CLL) and a concomitant IgA/lambda myeloma with phytohemagglutinin (PHA), lipopolysaccharide of E. coli (LPS) and S. aureus strain Cowan I bacteria (Cowan). A second malignant clone with three additional markers was observed after stimulation with Concanavalin A (Con A). A chromosome 14 aberration, inv(14) (q11-q32), may be of significance in the T-cell malignancy and may suggest a common clonal origin of the two tumors.

Aged↗

Signature of B-CLL with different prognosis by Shrunken centroids of surface antigen expression profiling.

With the aim of identifying the immunophenotypic profile of B-cell chronic lymphocytic leukemia (B-CLL) subsets with different prognosis, we investigated by flow cytometry the expression of 36 surface antigens in 123 cases, all with survivals. By analyzing results with unsupervised (hierarchical and K-means clustering) algorithms, three distinct immunophenotypic groups (I, II, and III) were identified, group I (51/123) with longer survivals, as compared to the group II (36/123) and III (36/123). The immunophenotypic signatures of these groups, as determined by applying the nearest Shrunken centroids method as class predictor, were characterized by the coordinated and differential expression of 12 surface markers, that is, group I: above-average expression of CD62L, CD54, CD49c, and CD25, below-average expression of CD38; group II: above-average expression of CD38, CD49d, CD29, and CD49e; and group III: below-average expression of the above markers, overexpression of CD23, CD20, SmIg, and CD79b. As opposed to groups II-III, group I B-CLLs lacked expression of ZAP-70 and activation-induced cytidine deaminase in the majority of cases, while more frequently had mutated IgV(H) genes and IgV(H) mutations consistent with antigen-driven selection. Our findings contribute to improve the immunophenotypical identification of disease subsets with different prognosis and suggest a set of surface antigens to be employed as prognosticators in routine diagnostic/prognostic procedures.

Adult↗

Patient accrual and interim statistical analysis in long-term randomized clinical trials: the French chronic lymphocytic leukemia CLL 80 protocol as a case study.

The CLL 80 clinical trial of treatment for chronic lymphocytic leukemia was designed using a three-stage prognostic classification. Within each stage patients received one of two treatments allocated by randomization, two adjacent stages having a common treatment. The fixed-sample design required 850 patients entered over a period of 8.5 years. Active patient recruitment led to 741 patients being randomized in four years. The first interim analysis, carried out at this point, showed a treatment difference in one stage which was statistically significant (p = 0.001) and justified an early termination of the protocol. This paper discusses practical aspects of patient accrual and interim analysis in this study.

Antineoplastic Combined Chemotherapy Protocols↗