Heteroantisera against a blast cell antigen.
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Biomedical subjects
Publications and source records attributed to R Billing.
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Fifteen lymphoblast cell lines, including B cell, T cell, Null cell, and myeloblast cells, were examined for the production of human neutrophil migration-inhibition activity. Only one T lymphoblast cell line established from a patient with hairy cell leukemia produced a neutrophil migration-inhibition factor spontaneously and after stimulation with Con A and PHA.
We have used in vitro immunotherapy before autologous bone marrow transplantation for three patients with acute lymphoblastic leukemia (ALL). Bone marrow was removed during remission, and mononuclear cells were separated by density-step centrifugation on Ficoll-Hypaque. The cells from each patient were treated with a heteroantiserum and complement to eliminate leukemic cells and were cryopreserved. Following chemotherapy and total body irradiation, the treated marrows were thawed and infused. All the patients showed positive evidence of returning marrow function before death. One patient who survived 4 months showed no evidence of leukemia at post mortem, and marrow sections demonstrated active hematopoiesis of all cell lines.
Recent studies using cytotoxic and cell separation techniques have identified differentiation-related antigens on human hematopoietic cells. These results, combined with bone marrow transplantation studies, have yielded a picture of antigenic modulation from the pluripotent hematopoietic stem cell through the differentiated end cells.
Normal human bone marrow contains cells capable of forming myeloid colonies (CFU-DG) in fibrin clots in diffusion chambers placed in the peritoneal cavity of cyclophosphamide-treated mice. Evidence has accumulated indicating that these colony-forming cells represent an earlier stem cell than the granulocyte-monocyte precursor cell (CFU-C) assayed in soft agar. We provide data showing that these stem cells express the "Ia-like" or DR antigen. In the presence of rabbit Ia antiserum at a titer of 1:300, all CFU-DG were inhibited. Cytotoxicity was complement-dependent. Data are also presented that suggest that the megakaryocyte stem cell also expresses the Ia antigen.
The sera of 233 kidney transplant patients before transplantation were tested by cytotoxicity against a panel of B and T lymphocytes at 5 degrees C and 37 degrees C. The results divided the patients into four groups: those whose sera reacted with B lymphocytes at 5 degrees C; those reacting with B lymphocytes at 5 degrees C and 37 degrees C; those reacting with T lymphocytes at 37 degrees C; and those with no antibodies. The patients with pre-transplant antibodies reactive with B lymphocytes at 5 degrees C had a significantly higher kidney-transplant survival rate at 6 months (70%) and 1 year (65%) than patients who had no antibodies (47% and 46%, respectively). Patients with antibodies reactive at 37 degrees C had a 6-month survival-rate of 38% when reactive against B cells and 43% when reactive against T lymphocytes. The cold cytotoxins were IgM.
Antisera have been raised in rabbits to the lymphoblastoid cell line NALM 1 precoated with anti-lymphocyte serum (ALS). Following absorption with chronic lymphocytic leukemia cells (CLL) the antisera reacted mainly with acute lymphocytic leukemia (ALL) cells, and were very similar in specificity to antisera raised to ALL cells precoated with ALS. Leukemia cells from the following numbers of patients were positive for the anti-NALM 1 sera in a complement-dependent cytotoxicity test; 11/14 ALL, 3/15 acute myelocytic leukemia (AML), 1/5 chronic myelocytic leukemia (CML) and 0/8 CLL. Normal B and T peripheral blood lymphocytes were negative. The titer of the anti-NALM 1 sera against positive cells was 1:64 to 1:256 whereas the undiluted sera did not react with negative cells. Ten out of 11 of the positive ALL cells were of the non-B non-T type. However, cells from 1/4 T ALL patients and a cultured T ALL line 8402 were also positive. Six of 12 cultured lymphoblastoid cell lines were positive, all of which were of malignant origin. The molecular weight of the ALL antigen detected by anti-NALM-1 serum was determined by immunoprecipitation and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) to be approximately 98,000 daltons.
Highly enriched preparations of monocytes, B and T lymphocytes, and granulocytes from 18 normal donors were serotyped in parallel in a complement-dependent cytotoxicity assay using allogeneic and heterologous antisera defining three independent tissue antigen systems. HLA and B-lymphocyte tissue antigens were detected on human monocytes although granulocyte antigens were absent. By cytotoxicity testing the presence of Ia-like antigens on monocytes was significantly diminished compared to the autologous B-lymphocyte population and has important implications in B-lymphocyte serology. The study indentified a number of human antisera obtained from multitransfused subjects and pre- and post-transplant organ recipients that were non-HLA and appeared to define monocyte-associated antigens. The serological implications of surface antigen expression on human monocytes compared with other peripheral blood cells are discussed.
Normal human granulocyte alloantigens were found on chronic myelogenous, acute myeloblastic leukemia cells, and a cell line of chronic myeloblastic origin (K562). Antigens were detected by human antisera positive for normal peripheral blood granulocytes but devoid of HLA activity. Very few acute lymphoblastic leukemia cells reacted positively, and none of the chronic lymphocytic leukemia cells seemed to bear granulocyte surface antigens. The recognition of these normal tissue isoantigens on myeloblastic leukemia cells is a necessary prerequisite for the identification of "leukemia-specific" of "leukemia-associated" antigens.
The participation of cell subpopulations in the expression of leucocyte migration inhibition factor (LMIF) in response to Concanavalin A and Protein A was evaluated for cells isolated from the peripheral blood of five healthy subjects. LMIF activity could not be attributed to the function of T cells, B enriched cells, or monocytes acting alone, or to a combination of B enriched cells and monocytes. The LMIF response was the result of a collaborative event that occurred between T cells and B enriched cells, or between T cells and monocytes.
Antisera were raised in New Zealand White rabbits against non-B, non-T acute lymphocytic leukemia (ALL) cells coated with antilymphocyte serum. Following minimal absorption with chronic lymphocytic leukemia (CLL) cells, the antiserum reacted mainly with non-B, non-T ALL cells. The following numbers of patients had leukemia cells that reacted with the ALL antisera: 13 of 18 with ALL, 3 of 27 with acute myelocytic leukemia, 1 of 8 with chronic myelocytic leukemia (CML), and 0 of 12 with CLL. The positive CML was a patient in CML blast crisis. Normal peripheral blood B- and T-lymphocytes and normal bone marrow were negative. Reactions of the anti-ALL serum (136K) were compared with the reactions of a rabbit anti-B-cell antiserum (63K) that reacted with approximately 70% of leukemia cells. Cultured lymphoblastoid cell lines from normal donors were negative by both cytotoxicity and immunofluorescence tests. However, by immunofluorescence testing, 8 of 17 known malignant lines from a variety of lymphoproliferative disorders were positive; 4 of these lines were of T-cell origin. By immunoprecipitation and polyacrylamide gel electrophoresis, the ALL antigen appeared to consist of a single polypeptide chain of approximately 98,000 daltons. The anti-ALL antiserum was not cytotoxic for normal myeloid stem cells (colony-forming units).
Antisera prepared against papain-digested spleen cell membranes were known to by cytotoxic for normal and neoplastic human B lymphocytes and for a majority of acute and chronic myeloid leukemic cells. It is now shown that these antisera are also cytotoxic for normal myeloid stem cells (CFU-C), thus providing a probable explanation for their occurrence in myeloid neoplasia.
Hybrids between two human lymphoma lines, Raji and Daudi (8A) and Raji BJAB (83) were examined for genetically determined and/or differentiation-related surface markers. HL-A B ce-l alloantigens, Fc and complement receptors, EBV receptors and beta2 microglobulin showed an autonomous ("co-dominant") expression in the hybrid. This is in contrast to most previous studies on other differentiation markers, involving as a rule crosses between cells of different lineages, where the differentiated pattern usually became "eclipsed" in the hybrid. Staining of activated complement and complement consumption tests showed intermediate or partially suppressed expression in the hybrids. This may be viewed in relation to the fact that these reactions do not merely depend on complement binding to the receptor, but also on subsequent activation and binding of the activated complement. A more complex interaction is also suggested for immunoglobulin production. Surface immunoglobulin showed a suppressive or intermediate pattern in both hybrids, whereas intracellular kappa chain production showed an amplification in the 83 hybrid. The beta2 microglobulin deficiency of the Daudi parent was corrected in the Raji/Daudi hybrid. Two new HL-A specificities,A10 and BW17, appeared on this hybrid which were not present on the parental lines. This suggests that the HL-A deficiency of the Daudi cell is due to its lack of beta2 microglobulin.
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The majority of human lymphocytic and myelocytic leukemia cells express a polymorphic antigen that is found on peripheral blood B-lymphocytes and cultured lymphoblastoid B-cell lines. These B-lymphocyte antigens were detected by 34 human alloantisera that were repeatedly absorbed with pooled platelets to remove all activity against HLA antigens and T-lymphocytes. Absorption studies indicated that a common antigen was present on both B-lymphocytes and positive leukemia cells. Leukemia cells could be subdivided into two groups based on the presence of the B-lymphocyte antigen. Fourteen of 18 acute myelocytic leukemia cells, 10 of 13 acute lymphoblastic leukemia cells, 4 of 6 chronic myelocytic leukemia cells, and 2 of 2 chronic lymphocytic leukemia cells were positive. This group of leukemia cells also reacted with rabbit anti-B-cell sera raised to papain digests of spleen cell membranes. F(ab')2 fragments of the rabbit antsera were shown to specifically block the reactions of the human antisera against B-cells and leukemia cells. These results suggested that the rabbit and human anti-B-cell sera were reacting with identical molecules. This conclusion was supported by immunoprecipitation data.
The effect of sera from 256 human cancer patients on the human antibody-dependent cellular cytotoxicity (ADCC) assay was studied. The cancer sera were compared to normal sera for their ability to alter the effector function of lymphocytes following 30 minutes' treatment at 37 degrees C. Even with this brief treatment, 74% of the 256 cancer sera inhibited effector activity. In most instances this inhibition was greater in patients with metastatic disease than in those without. Patients with colon and prostate cancer showed a statistically significant increase in inhibition among patients with disseminated disease (P less than 0.02 and P less than 0.01, respectively). An opposite effect was noted only in melanomas. It is suggested that the inhibition of the effector function in ADCC is a potential in vitro measure of the immunologic status of cancer patients.
Both F(ab')2 fragments and intact antibody from rabbit anti-B-cell antisera were shown to specifically block the mixed lymphocyte culture (MLC) reaction. The antisera, which were raised to papain-solubilized spleen cell membrane antigens, appeared to block the stimulator cell and not the responder cell. Specificity of the blocking was shown in that antisera to beta2 microglobulin would not block the MLC-stimulating cell. Membrane antigens from acute lymphocytic leukemia cells were labeled with 125I, solubilized with sodium deoxycholate and immunoprecipitated with either human or rabbit anti-B-cell antisera. Both sources of antisera precipitated polypeptide subunits of 27,000 and 35,000 daltons.
Two EBV-negative lymphoma lines of human B-cell origin, BJAB and Ramos, were compared with altogether six of their in vitro EBV-converted, EBNA- and EBV-DNA-carrying sublines (four of Ramos and two of BJAB derivation). All converted lines closely resembled the parental line with regard to karyotype and HL-A and B antigen typing. Induction of EBV antigens (EA and VCA) by P3HR-1 virus superinfection was either similar in the converted and the negative lines, or somewhat increased in certain converted lines. These findings argue against a simple, virally determined repressor model and emphasize the role or cellular controls in restricting the EBV cycle in virus-carrying B-lymphocyte lines of human origin. IUdR inducibility varied in the different converted lines. There was a possible relationship between average number of EBV-genome equivalents per cell and inducibility. Converted sublines did not differ from the original negative lines with regard to surface immunoglobulin and Fc receptors. There was a dramatic increase in complement-consuming ability, however, following EBV conversion. Among the EBV-positive lines, there was a linear relationship between complement-consuming and EBV-receptor activity, the latter measured by a quantitative absorption test.