Immunohistochemical demonstration of cell surface antigens on tissue sections of lymphomas.
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Biomedical subjects
Publications and source records attributed to S Thierfelder.
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A series of 66 children with acute lymphoblastic leukemia (ALL) at diagnosis were investigated (simultaneously) for various surface markers. For this purpose the reaction of specific antisera against ALL antigens and T cell antigens was analysed in every case by several test systems, namely immunofluorescence, microcytotoxicity and complement fixation. A clearly defined classification in 6 subgroups of ALL emerged. The clinical data at presentation and possible correlations with the immunological subgroups were demonstrated.
Human lymphocytes derived from various central and peripheral sources were separated on linear density gradients (LDG). Cells from individual density fractions were tested in parallel for: the capacity to form nonimmune rosettes with neuraminidase-treated SRBC, the number of surface-associated HTLA, and in vitro proliferative responses to mitogenic lectins and alloantigens. Heterogeneous density distribution profiles were obtained for all sources of human T cells and revealed an organ specificity. The various T cell density classes obtained from identical organs as well as the identical density classes of different sources revealed to some extent differences in their surface marker patterns and/or their in vitro reactivities. On the basis of the combined techniques at least two major subsets among thymocytes were identified that differed in both surface properties and functional capacities. Density classes of T cells from all peripheral sources were distinguished from thymocytes by a homogeneous lowered HTLA expression. Whereas clear-cut differences in the in vitro functional capacity were observed between the two thymocyte subsets, less striking but still significant differences were found to exist among the various density classes of peripheral T cells.
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Acute lymphatic leukaemia (ALL) in infants is a very heterogeneous disease from the immunological point of view. Phaenotyping of leukaemic cells in 138 patients via different immunmarkers has so far allowed subdivision into 6 subgroups. Clinical risk factors and correlations with the immunologically defined subgroups, as well as their significance for prognosis, are demonstrated with the help of 61 examined ALL patients. 90-95% of all acute lymphatic leukaemias can presently be identified via heterologous antisera. After corresponding absorption, these antisera show high cytotoxicity against ALL cells without displaying any aggressivity towards normal haemopoetic stem cells. The authors make therapeutic use of anti-T cell and anti-common ALL (cALL) globulin in autologous bone marrow transplantation. This method involves withdrawal of bone marrow from patients with ALL in complete remission, the marrow being then fractionated, treated with antiserum to destroy residual leukaemic cells, and then cryopreserved. The leukaemia-free bone marrow will thus be available for transplantation in relapse.
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Cells of the Reh line, originally derived from an ALL of the 'Non-T, Non-B' type were cultured in diffusion chambers implanted intraperitoneally into perirradiated CBA mice. At different intervals over a period of 20 d changes in surface characteristics were examined by labelling the cells with AcALLG, ATCG as well as with polyvalent AIg. The evaluation was performed by using direct immunofluorescence. In addition, the ability to form rosetts with SRBC, AET-treated sheep erythrocytes, mouse red blood cells and EoxAC was tested. On day 0 of the diffusion chamber culture the cells only carried cALLA, and no rosette formation was observed. In the course of the diffusion chamber culture the cells unequivocally developed T-cell antigen, and in 1 of 2 experiments they further acquired a receptor for forming AET- and E-rosettes. Conversely, in the other experiment a receptor for mouse red blood cells was detected in a considerable portion of the cells. Our data show that the rather undifferentiated Reh line cells in vitro are able to develop features of mature T-cells and attributes of early B-cells during the diffusion chamber culture. In vitro they apparently retain a bivalent potentiality of lymphatic maturation. The diffusion chamber system proves to be a suitable tool for promoting differentiation in these cells.
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The specificity of an antiserum against ALL-cells lacking B- and T-cell markers was characterized. The antiserum was used in several indicator systems to investigate the expression of leukaemia-associated antigens on normal lymphocytes, lymphoid cell lines and various leukaemia cells. The relevance of such antisera for the detection and classification of leukaemic cell populations is discussed.
The lymphoblasts from 100 patients with acute lymphocytic leukaemia were investigated for the expression of receptors for sheep erythrocytes (E) and of a specific heterologous T cell antigen (T). In 17 cases, both T cell markers were expressed simultaneously on the leukaemic cells. In 13 cases only T antigens could be demonstrated on the lymphoblasts. A quantitative analysis of T antigens by immunoautoradiography revealed that the T expression of E-T+ -lymphoblasts was in general like that of E+T+-lymphocytes in the blood of normal persons, in several cases even higher. Therefore, the failure of E-rosette formation cannot be correlated to a decrease of the other T cell differentiation marker. In 7 out of 9 tested cases, a strong acid phosphatase reaction product located paranuclearly could be demonstrated. Complement-receptors were expressed in 3 of 5 cases which were also demonstrated in some cases of the E+T+-ALL group. The latter group was characterized by a T antigen expression like that of thymocytes. 4 cases of the E-T+ALL group were adults. Since the leukaemia cells of 2 cases were negative for acid phosphatase, PAS and all surface markers including cALL antigen, the T antigen can classify undifferentiated and otherwise unclassificable leukaemias. The clinical signigicance of the E-T+-ALL seems to be important since 5 out of 9 children with this type of ALL died soon after diagnosis.
The cellular immune status of 48 canine chimeras was studied between 15 days and 3 years after a lethal dose of cyclophosphamide (CY) and an allogeneic marrow graft. Marrow graft recipients were given no further immuno-suppressive treatment. Recovery of total white blood cell counts was complete by day 20 after grafting. Peripheral blood lymphocyte counts did not reach the normal range until day 100. Short-term chimeras (less than 100 days postgrafting) showed marked impairment of cell-mediated immunity as indicated by the inability to respond adequately to nonspecific mitogenic (PHA, ConA, PWM) and specific antigenic (allogeneic lymphocytes, PPD) stimulation in vitro. Skin test reactivity to PPD after immunization with BCG was also significantly depressed. Long-term chimeras (more than 100 days postgrafting) had normal in vitro response to mitogens; lymphocyte blastogenesis to specific antigens however remained decreased for prolonged periods postgrafting. The survival time at first set allogeneic skin grafts showed no significant difference between chimeras and normal control dogs. Lymphocyte transformation in response to stimulation with specific antigens may prove a useful parameter for the in vitro evaluation of defects in cellular immune reactivity following allogeneic marrow transplantation.
Acute lymphoblastic leukemia (ALL) in childhood is not a homogeneous disease. By membrane phenotyping 6 groups of ALL in 66 children could be differentiated: Group I (38%) common ALL (anti-cALL+. Group II (32%) common ALL with T cell marker in addition (anti-cALL+, anti-T+). Group III (14%) T. cell ALL with E-rosette marker (anti T+, E+). Group IV (9%) T cell ALL without E-rosette marker (anti T+, E-). Group V (6%) undifferentiated ALL (anti-cALL-, anti-T). Group VI (1%) B cell typ (SmIg+). 19 or 20% of children in groups I and II opposed to 67 or 100% of children in groups III and IV had a white cell count at diagnosis of greater than 25,0 x 10(3)/mm(3) Mediastinal involvement was typical for groups III and IV. No significant difference was found so far for age or sex in the groups.
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Hematological and immunosuppressive effects of various single doses of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) were evaluated in the preclinical canine model. Immune function appeared not to be impaired by treatment with CCNU, but severe myelotoxicity contributed to the death of 3 of 4 dogs given 10 mg CCNU per kg and 5 of 5 dogs given 15 mg CCNU per kg within 10 days after drug administration. Infusions of autologous bone marrow protected 6 of 6 dogs receiving 15 mg CCNU per kg and 6 of 6 dogs receiving 20 mg CCNU per kg from lethal marrow failure. Dogs given 30 mg CCNU per kg and autologous marrow died within 7 days from severe gastrointestinal toxicity. We conclude that autologous bone marrow support may allow the use of high-dose CCNU regimens and thereby increase its therapeutic efficacy in the treatment of advanced cancer.
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