Sixty-two explants from peripheral blood, bone marrow and cerebral fluid of children with acute lymphoblastic leukemia (ALL) and leukemic transformed non-Hodgkin lymphoma (NHL) were cultivated for at least 8 weeks. Although lymphatic cells persisted up to 16 weeks in tissue culture, no proliferation was observed in 54 cultures. From the remaining cultures, eight permanently growing cell lines were obtained. Five of these were EBNA (Epstein-Barr virus-specific nuclear antigen)-positive. Three, however, were ENBA-negative and lacked Epstein-Barr virus genomes. Two cell lines (KM-3 and SH-2) expressed neither B nor T cell characteristics. One line (JM) expressed T cell characteristics and complement receptors. The growing lymphatic cells represented leukemic cells, since the pattern of cytochemical staining and that of membrane receptors of lymphoblasts from the same donor prior to cultivation were identical. All leukemic cell lines were derived from patients in relapse. In contrast, no proliferation of leukemic cells occurred in explains from patients revealing the first manifestation of the disease. These results suggest enhanced growth potential of lymphoblasts resisting antileukemic therapy.
A T-cell marker was expressed by the leukemic lymphoblasts of a 14 years old boy. After 14 days of cultivation, a permanently growing lymphoblastic cell line JM, which manifested the T-cell marker over the whole period of subcultivation, was established from the blood. In vitro synchronization experiments showed that this important cell marker in the G1 phase of the cell cycle is expressed on the surface of the blasts to a reduced degree, compared to the level in the other cell cycle phases.
Immune complexes (ICs) participate in the pathogenesis of various diseases and can be shown in 18% of all hospitalized patients (excluding those with infectious diseases) by means of a sensitive method such as the Raji-cell radioimmune assay. However, before this test can be applied to quantify disease activity in renal, connective tissue and neoplastic diseases, it must be recognized that febrile infections of the upper respiratory tract also induce ICs in 86% of all patients. The immune complexes containing microbial antigens can be reduced or removed by a single injection of human immunoglobulin. This is a simple method to distinguish between the immune complexes of different specifities. The resulting removal of some immune complexes may be the explanation for the claimed therapeutic effect of gammaglobulin injection in normogammaglobulinemic patients.
The T-and B-lymphoeytes of 17 patients with immunodeficieney diseases were determined quantitatively. Simultaneously, their functional behavior, i.e. the response to stimulation by phytohemagglutinin and the immuneglobulin levels were studied.In a preliminary study devised to check the accuracy of the methods used, the cell markers (rosette formation with sheep erythrocytes for T-cells and membrane immunefluorescence for B-cells) were defined by means of two different lymphoid cell populations: as nearly pure T-cells the fresh suspensions prepared from thymie biopsies of 3 young children were used, as B-cell rich preparations the isolated lymphocytes of 3 patients with chronic lymphatic leukemia.Immunodeficieney syndromes were classified according to the recommendations of a World Health Organization expert committee [12,13]: 2 patients with agammaglobulinemia, 5 with variable immunodeficieney, 4 with ataxia teleangiectatica, 3 with severe combined immunodeficiency and 3 with Wiskott-Aldrieh syndrome were extensively studied ( Table 2). The results are relevant for the better understanding of norreal differentiation and its disturbances leading to the different deficiency syndromes (el. Fig. 1).
The binding of 12 different fluorescein-conjugated lectins to 10 ALL (acute lymphoblastic leukemia) cell lines and two cell lines derived from patients in blast crisis of myeloid leukemia was examined. The specificity of the membrane fluorescence was demonstrated by inhibition with various saccharides. Three of the lectins bound to all cell lines, four bound to only some of the lines, and five were not bound. There was no correlation between the binding pattern and the immunological phenotype of the cultured lymphoblasts. The lectin from Lens culinaris, however, in the experimental condition used (incubation at 4 degrees C, fluorescein conjugation at pH 8.5), bound only to the cell membranes of 'Ia-like antigen, positive cell lines. Although three lectins (Lens culinaris, Pisum sativum, concanavalin A) had an identical monosaccharide specificity, they bound to different cell lines. Membrane fluorescence with the lectins from Helix pomatia, Arachis hypogea, and Ricinus communis (MW 60,000) was achieved after treatment with neuraminidase. It was shown that binding of the lectins from Helix pomatia and Ricinus communis 60 was effected by enzymatically exposed glycoproteins, whereas the lectin Arachis hypogea was bound via neuraminidase which stuck to the cell membrane.
B-Cell-allo-antigen was demonstrated on lymphoblasts of children with acute lymphoblastic leukemia and on 14 permanent growing lymphoblastoid cell-lines. Pooled sera of 4 selected multiparas and a rabbit antiserum were used as specific antisera. Antigen was more effectively detected with heterologous antibodies than with human antisera. Primary lymphoblasts were characterized by B-cell-allo-antigen and lack of membrane immunoglobulins and Fc-receptors on the same cell. The quality of remission, i.e. the early diagnosis of relapse was better surveyed by immunological characterization than by cytological assessment. The bone marrow was examined in 8 patients with initial leukemia, in 13 patients with partial remission and in 30 patients in complete hematological remission. A striking discrepancy between immunological and cytological examination was observed in the bone marrow 4 weeks after inital anti-leukemic chemotherapy. At that time an increased number of blast-typed cells could still be counted in the immunological evaluation, although the cytological evaluation revealed already a complete hematological remission.
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