T-lymphocytes bearing Fc-receptors for IgG in B-cell chronic lymphocytic leukaemia

Peripheral venous blood lymphocytes of 10 untreated B-CLL patients at stages Rai 0-III and of 10 healthy individuals were studied. In B-CLL the absolute number of T lymphocytes present in peripheral blood was similar to that in healthy controls; however, the ratio of T lymphocyte subpopulations in B-CLL was changed in favour of TG cells, the absolute number of which was found to be increased. In B-CLL DNA synthesis of the TG cells in response to PHA stimulation was more intense than in healthy individuals. These observations are discussed in the light of the conflicting reports on the numerical and functional changes in B-CLL T-cells.     

Identification of cellular immunoglobulins in chronic lymphocytic leukaemia by immunoperoxidase staining.

An indirect immunoperoxidase technique has been used for visualisation of cellular immunoglobulins in chronic lymphocytic leukaemia. Baker's formol calcium was used as fixative. Monoclonal light and heavy chain patterns were demonstrated in 24 out of 27 cases. Only one case did not have any demonstrable immunoglobulins. The presence of alpha or gamma heavy chain immunoglobulin isotypes in leukaemic lymphocytes was found to be related to low mouse rosetting capacity (p less than 0.05).     

Autoimmunity in chronic lymphocytic leukaemia.

The prevalence of autoantibodies in B cell chronic lymphocytic leukaemia (B-CLL) was investigated. A lower prevalence of autoimmune haemolytic anaemia than that found in other series was found: large numbers of non-progressive stage A disease cases were included, in which the prevalence of autoimmune haemolytic anaemia is low. Non-haematological autoantibodies were no commoner than in age matched controls. Whatever explanation is offered for autoimmune phenomena in B-CLL it must take account of the fact that those phenomena are virtually confined to autoantibodies against the formed elements of the blood.     

Prolegomenon for chronic lymphocytic leukaemia

Chronic lymphocytic leukaemia (CLL) is a unique lymphoproliferative disorder that scarcely occurs under the age of 40; thereafter the incidence of CLL increases exponentially with age. CLL is characterized by progressive expansion of malignant CD5+ME+ B-cell clone accompanied by a myriad of cellular and humoral immune defects. Each of them might be linked to different clinically manifested complications such as increasing rate of infections, autoimmune disorders and disturbed immune surveillance against tumour cells. We assume that CLL occurs as a consequence of age-dependent, genetically related functional restrictions of the thymic microenvironment in supporting common lymphoid progenitor cells (CD5+ME+CD4-CD8-) to differentiate into mature T-cell and B-cell descendants. In conjunction with genetic abnormalities developing in B-cell progenitors, presumably expressing P glycoprotein (Pgp+), we postulate that developmentally altered T-cell descendants, along with quantitative imbalance among CD4+, their subsets and CD8+ lymphocytes in the peripheral blood, play an important additional role in facilitating the malignant B-cell clone emergence and in modulating the CLL clinical evolution. Namely, imbalance of any of T-cell-mediated cell interactive homeostatic mechanisms accompanied by imbalance in the production of various cytokines might in CLL influence leukaemic B-cell growth by deregulating inducer (c-myc and p53) and/or suppressor (bcl-2 and mutant p53) oncogenes responsible for the promotion or suppression of B-cell mitogenesis that may in turn further contribute to their impaired differentiation and/or differentiation arrest. In conclusion, CLL might be interpreted as a primary immunodeficiency syndrome developing in elderly population due to gradually evolving restriction of genetically controlled programs in the thymic microenvironment responsible for irregular maturation of common lymphoid progenitor cells that constitutively express CD5 antigen and ME receptor into T-cell and B-cell descendants.     

Cell surface localized IgM-kappa immunoglobulin reactivity in a case of chronic lymphocytic leukaemia

The cell surface of peripheral lymphocytes from a patient with chronic lymphocytic leukaemia reacted strongly with fluorescein conjugated anti-IgM and kappa serum. Moreover in the presence of complement anti-IgM serum was cytotoxic. This patient was unusually resistant to various therapeutic measures. Serum immunoglobulin levels were slightly subnormal. The reactivity described differs from other chronic lymphocytic leukaemias studied but could be found in a number of Burkitt cases. The leukaemia cells may represent the malignant transformation of a normal lymphoid cell specialized to carry immunoglobulin on its cell membrane.     

Advances in therapy of chronic lymphocytic leukaemia

Chronic lymphocytic leukaemia (CLL) is a lymphoproliferative disorder characterized by the progressive accumulation of clonal small mature-looking lymphocytes, usually of B cell origin. In addition to a better understanding of many biological features, during the last 10-15 years a great progress has been made in the prognostic characterization of the disease. The therapeutic achievements, however, have been less impressive. The possibility to identify patients with different prognosis has renewed interest in the treatment of this disorder. A considerable number of controlled trials have been performed or are in course, and new perspectives for CLL treatment are emerging. Basically four different kinds of measures are used: 1) chemotherapy, 2) radiotherapy, 3) adjuvant measures and 4) new modalities.     

Non-cytotoxic antibodies in chronic lymphocytic leukaemia.

Non-cytotoxic Fc receptor blocking antibodies against autologous B lymphocytes were sought in sera from patients with chronic lymphocytic leukaemia (CLL), using a rosette inhibition assay. They were found in 11 of 52 (21%) of patients with CLL, but were not associated with previous blood transfusion or pregnancy, suggesting that they were unlikely to have resulted from allogeneic stimulation. Fc receptor blockade was more commonly detected in sera from patients with stage B rather than stage A CLL (Binet classification), though this did not achieve significance beyond the 90% level, and it was noted in 62.5% of those with lymphocyte doubling times of less than one year, compared with 36.3% of those whose lymphocyte doubling time was more than one year. The results indicate that autologous Fc receptor blocking antibody activity occurs in sera from patients with CLL, is more likely to be generated by the tumour itself than by allogeneic stimulation, and is associated with increased tumour load. Such antibodies may permit tolerance of tumour by the host.     

Karyotypic evolution in B-cell chronic lymphocytic leukaemia

Sequential cytogenetic studies were performed on a minimum of two and a maximum of nine occasions (mean 3.6) on the peripheral blood leucocytes of 112 patients with B-CLL. On initial cytogenetic analysis, 58 had a normal karyotype and 64 had a clonal abnormality. Karyotypic evolution occurred in 18 patients (16%). There was no significant difference in the incidence of disease progression between patients with a stable karyotype and those who underwent karyotypic evolution. In only one patient was there a clear association between disease progression, a change in cell morphology and karyotypic evolution.     

Serum paraproteins in chronic lymphocytic leukaemia.

The presence of paraproteins in the sera of 10 patients with chronic lymphocytic leukaemia (CLL) was investigated using immunoisoelectric focusing. Monoclonal immunoglobulins were found in nine of these 10 sera. Five sera contained a single monoclonal IgM paraprotein, one serum contained a single monoclonal IgG paraprotein, while three sera contained more than one monoclonal paraprotein--namely, IgM + IgD, IgM + IgG, and IgM + IgD + IgG. The results indicate that the malignant B cells of CLL may be at a later stage of differentiation than previously assumed and serum monoclonal immunoglobulin could be of value as a tumour marker.     

Cytoplasmic inclusions in lymphocytes of chronic lymphocytic leukaemia

Summary Peripheral blood from 90 CLL patients was examined by light- and electron-microscopy for the occurrence of crystalline inclusions in lymphocytes. Inclusions were demonstrated in 10 patients (11%). In these patients the inclusions were present in 5–45% of peripheral blood lymphocytes.In the light microscope the inclusions appeared as rectangular, unstained structures in May-Grünewald Giemsa and PAS stains. In the electron microscope the inclusions appeared as intracytoplasmic, completely partially membrane-bound bodies, which were often associated with dilated profiles of rough endoplasmic reticulum. The ultrastructure of the inclusions was granular.In immunofluorescence staining the inclusions were found to contain immunoglobulin of the same type and class as the surface membrane-bound immunoglobulin of the neoplastic lymphocytes, most frequently IgMlambda. The lymphocytes of one case with kappa light chains at the cell surface membrane contained inclusions of the same ultrastructural morphology as those of the other cases with lambda light chains.The presence of inclusions was not associated with any specific clinical or prognostic features. The inclusions persisted during antileukaemic therapy. Their formation may be related to a dysfunction in the synthesis of surface membrane-bound immunoglobulins.     

Spontaneous lymphocyte transformation in chronic lymphocytic leukaemia.

In ten patients with CLL and in eight controls T lymphocytes were separated out by the rosetting method. The in vitro reactivity of these cells and of unseparated lymphocytes was studied in cultures without stimulants. A net increase of spontaneous lymphocyte transformation was found in the populations enriched with T cells in patients with CLL with respect to analogous populations in the controls. This increase is interpreted as a cell-mediated immunological reaction and due to T-cell sensitization towards the neoplastic B cells.     

Unreleased intracellular monoclonal macroglobulin in chronic lymphocytic leukaemia

In two patients presenting as chronic lymphocytic leukaemia, immunofluorescence studies have demonstrated the presence of an accumulated intracytoplasmic material reacting with antisera to μ chains and only to one type of light chains, whereas their serum contained neither monoclonal IgM nor free μ chains.

In the first case, the IgM lambda inclusion bodies were crystals detected in the cytoplasm of small lymphocytes and were not clearly lined by rough endoplasmic reticulum. IgM λ molecules were present at the surface of the lymphocytes.

In the other patient, who elaborated free κ light chains, the inclusions found in marrow plasma cells and in peripheral blood lymphocytes were Russell bodies.

The practical and theoretical importance of non-secretory immunoproliferative disorders, characterized by an unreleased monoclonal immunoglobulin marker, is outlined.

     

Serum beta-2 microglobulin in chronic lymphocytic leukaemia

Serum-beta-2-microglobulin was measured by radioimmunoassay in 25 patients with chronic lymphocellular leukaemia in stages III-IV according to the Rai classification. A significant positive correlation was found between the absolute lymphocyte count and the serum-beta-2-microglobulin level. No similar relationship was observed between the score indicative of organ infiltration and the beta-2-microglobulin valve. On the evidence of the results, the increased production of beta-2-microglobulin is attributed in the first place to the circulating lymphocytes. The assay has been found to provide a reliable indicator, suitable for the monitoring and evaluation of therapy.     

Intracellular IgA immunoglobulin crystals in chronic lymphocytic leukaemia

Lymphocytes from a case of chronic lymphocytic leukaemia were found to contain crystalline protein inclusions within cisternae of rough endoplasmic reticulum. These inclusions were characterized as IgA λ immunoglobulin and differed from the immunoglobulins found at the surface of the cells containing the inclusions. No serum paraprotein was detected in the serum of the patient, although small amounts of λ light chain protein were present in the urine. These observations suggest that the crystalline immunoglobulin was not excreted in significant amounts by the lymphocytes.