Monoclonal anti-CD4 in arthritis.
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Antibodies, Monoclonal↗
Biomedical subjects
W Knapp
Publications and source records attributed to W Knapp.
Neopterin as discriminating and prognostic parameter in healthy homosexuals, ARC and AIDS patients.
Urinary neopterin, a sensitive marker for activation of cell mediated immunity, is compared to clinical and laboratory data in 5 patients with AIDS, in 15 patients with ARC and in 40 male subjects without AIDS-related clinical signs attending an AIDS outpatient clinic. The sensitivity of neopterin for AIDS-related diseases was higher than that of the CD4+/CD8+ subset ratio. The differences in neopterin levels between the controls, ARC and AIDS patients were found to be more significant than the T-cell subset data. The observation that the 3 AIDS patients with the highest neopterin levels have died whereas the two others with the lower levels are still alive also underlines the prognostic potential of urinary neopterin level determinations. Measurement of urinary neopterin is thus recommended as an additional criterion for monitoring ARC and AIDS patients.
AIDS-Related Complex↗
[Atenolol poisoning].
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Alcoholic Intoxication↗
Bone-marrow purging with monoclonal antibodies and human complement in ALL and AML.
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Antibodies, Monoclonal↗
Leukemic colony-forming cells in acute myeloblastic leukemia: maturation hierarchy and growth conditions.
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Bone Marrow↗
In vitro culture studies of granulocyte/macrophage and erythroid progenitor cells in lymphoproliferative disorders.
Granulocyte/macrophage progenitor cells (CFU-GM) and erythroid progenitor cells (BFU-E) have been assayed in peripheral blood (PB) and/or bone marrow (BM) from 12 patients with acute lymphocytic leukemia (ALL), 16 patients with chronic lymphocytic leukemia (CLL) and 31 patients with various forms of non-Hodgkin lymphoma (NHL) without BM involvement. Progenitor cell growth in PB and BM from the NHL patients did not differ statistically from controls (p greater than 0.1). CFU-GM and BFU-E per ml PB were markedly increased in ALL and CLL patients (p less than 0.001) while CFU-GM and BFU-E per plated BM cells from these patients were severely depressed (p less than 0.001). Lymphoblasts from one ALL patient failed to inhibit CFU-GM and BFU-E-derived colony growth from control PB mononuclear cells. The high levels of circulating progenitor cells in ALL and CLL patients clearly distinguish them from other cytopenic hematological malignancies, in which decreased progenitor cell levels have been demonstrated previously (acute myeloid leukemia, hairy cell leukemia). The cause of this finding and its pathophysiological implication still remains to be established.
Bone Marrow↗
Immunoenzymatic staining methods for simultaneous demonstration of chromosomes and cell surface markers.
We have developed immunocytochemical staining methods for the simultaneous phenotypic and karyotypic characterization of individual cells. Following a mild hypotonic pretreatment, isolated cells are cytocentrifuged on poly-L-lysine coated slides, fixed in formol buffered acetone, and subsequently labeled with monoclonal antibodies utilizing indirect immunoenzymatic staining procedures with horseradish peroxidase (HRP) or alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) as second antibodies. Preparations are refixed consecutively in methanol and 45% acetic acid and counterstained with either "Stains-all" (HRP labeled preparations) or Giemsa (APAAP labeled preparations). C-banding or weak G-banding, which allows the identification of individual chromosomes, can be induced in labeled as well as unlabeled mitotic cells by Ba(OH)2 and/or 2 X SSC treatment after refixation, respectively. Our method has been successfully tested with a variety of monoclonal antibodies against lymphoid, myeloid, erythropoietic, and thrombopoietic cell surface antigens. It is fast, allows the adjustment of the intensity of cell surface staining, and results in permanent preparations suitable for light microscopic analysis.
Alkaline Phosphatase↗
Induction of human complement activation without cytolysis by mouse monoclonal antibodies to human leukocyte antigens.
Ten monoclonal antibodies to human leukocyte subsets that had previously been shown to lyse their respective target cells in the presence of rabbit serum as complement source were evaluated for their cytolytic capacity with human complement. Four of the ten were lytic with human complement. All were of IgM type. Antibodies were also evaluated for their capacity to induce C3 binding to target cells. With this method we could demonstrate that, indeed, 3 of the 6 noncytolytic antibodies had the capacity to initiate the human complement activation process and to induce C3 binding. Two of these 3 antibodies were of IgM class (VIT3 and VIM13), one of IgG3 (562). From the practical point of view the most interesting of these 3 antibodies is the nonmitogenic anti-CD3 pan-T cell antibody VIT3. Therefore, this antibody was analyzed in more detail. VIT3 antibody concentrations needed to induce detectable C3 binding to human T cells are very low (down to 1 ng VIT3/ml). Human serum as complement source can also be considerably (100X) diluted before C3 binding becomes undetectable. Activation of C3 is a prerequesite for VIT3-induced C3 binding, and bound C3 seems to lack the C3a fragment. Bound C3, in contrast to the quickly occuring antigenic modulation of the CD3 complex and the simultaneous disappearance of the antibody coat, remains expressed also after prolonged incubation at 37 degrees C. C3 fragments bound to T cells after activation with VIT3 are also recognized by cells bearing C3 receptors of types CR1 and CR2.
Antibodies, Monoclonal↗
Changes of the phenotype of erythroid progenitor cells (BFU-E, CFU-E) and their progenies during early steps of differentiation.
Early differentiation processes of human erythroid progenitor cells (BFU-e, CFU-e) have been studied during in vitro proliferation using a panel of monoclonal antibodies with known reactivity on different levels of the erythroid cell line. Two antibodies recognizing structures on BFU-e (VIP-2b, BMA 021), two antibodies reactive with CFU-e and nucleated red cells (5F1, CLB-Ery-3) and one antibody directed against glycophorin A (VIE-G4) were used for this study. Normal human bone marrow cells were induced to proliferation in an erythroid progenitor cell assay and, after different periods of incubation, agar cultures were treated with these antibodies and complement. Thereafter, the remaining erythroid cells were incubated again to continue their proliferation with the same stimulators as before. The changes of the phenotype of BFU-e and CFU-e progenies during in vitro proliferation were determined by the reduction of colony formation in comparison with untreated control cultures. Our results indicate that the loss of HLA-DR antigens and the p45 structure is accompanied by the acquisition of structures recognized by the antibodies 5F1 and CLB-Ery-3. After 5-7 d of incubation BFU-e derived progenies exhibit the same antigenic structure as has been found for CFU-e. Glycophorin A expression could only be demonstrated at a late differentiation stage of the erythroid cell lineage.
Antibodies, Monoclonal↗
Establishment of three permanent human leukaemia cell lines producing immunoreactive calcitonin.
Three permanent human leukaemia cell lines, designated EW2, LG3 and MS6, were established from bone marrow aspirates of a patient with acute myelomonocytic leukaemia (EW2), acute monocytic leukaemia (LG3) and acute myeloblastic leukaemia (MS 6). In vitro all lines exhibited a myelomonocytoid marker profile in terms of classical staining reactions and cell-surface antigen structure. In supernatants and extracts of EW2 and MS 6 cells immunoreactive human calcitonin was found in raised levels. No significant levels of immunoreactive human calcitonin were found in supernatants of LG3 cells, while extracts of LG3 cells and the patients sera at diagnosis contained high levels, suggesting the in vitro selection of a non-secreting clone of leukaemic cells. Gel-chromatography showed several peaks of ihCT with higher molecular weight than normal human calcitonin, suggesting the production of precursor molecules. In addition to ihCT raised levels of immunoreactive calcitonin gene related peptide (ICGRP) was found in supernatants of EW2. These data support the concept of ectopic immunoreactive human calcitonin production by leukaemic cells.
Antigens, Neoplasm↗
Transfer factor for adjuvant immunotherapy in cervical cancer.
In a prospective randomized double-blind study of 60 patients with invasive cervical cancer, 32 were treated with transfer factor (TF) derived from leukocytes of the patients' husbands, and 28 were treated with placebo. Within the first 2 years after radical hysterectomy, five out of 32 TF-treated patients and 11 out of 28 placebo-treated patients developed recurrence of malignancy. Excluding one further patient with intercurrent death this difference is significant (chi 2 = 3.9915; P less than 0.05). Subdividing the collectives, significant differences were found in patients aged below 35 years and in patients with stage I disease. Identical immune profiles were checked in leukocyte donors prior to leukophoresis and were serially checked in patients. Antigen-specific correlations were found between donors' and recipients' reactivities but not between donors' reactivity and recipient's course of the disease.
Clinical Trials as Topic↗
Vim-2, candidate monoclonal antibody for purging autologous marrow grafts in acute myeloblastic leukaemia.
In this report we present a method for purging acute myeloid leukaemia progenitors (AML-CFU) from autologous marrow grafts. Previous studies using flow cytometry and cell sorting had indicated that the monoclonal antibody (MCA) Vim-2 reacts strongly with AML-CFU in a significant number of cases, whereas the majority of normal bone marrow precursors are Vim-2 negative. To investigate the applicability of Vim-2 for selective purging, we performed complement-dependent cytotoxicity experiments and determined the specific lysis of AML-CFU. In 11 of 17 patients, 97.5 +/- 4% (mean +/- SD) of AML-CFU were killed following immune-mediated lysis. Multi-lineage normal haematopoietic progenitors (CFU-GEMM) were not significantly affected by the MCA Vim-2-mediated complement lysis. These data suggest that the MCA Vim-2 is a useful reagent for the elimination of residual AML progenitor cells from remission bone marrow in patients subjected to autologous marrow transplantation.
Antibodies, Monoclonal↗
Interleukin 2 receptors on cultured murine epidermal Langerhans cells.
Rat monoclonal antibodies 3C7 and 7D4 detect two distinct functional regions of the murine interleukin 2 (IL 2) receptor. When studying the emergence kinetics of IL 2 receptors in mixed epidermal cell (EC)-lymphocyte cultures by using 3C7 and 7D4 in an indirect immunofluorescence assay, we regularly encountered a distinctive membrane fluorescence not only on lymphocytes, but also on a subpopulation of cells exhibiting a dendritic morphology. Reasoning that these 3C7/7D4-reactive dendritic cells might represent a subpopulation of epidermal dendritic cells, we studied mouse EC for the presence of 3C7/7D4- reactive cells. Although 3C7/7D4 reactivity was never detected on freshly isolated EC or on epidermal sheets, a small number of 3C7/7D4+ cells was encountered after 24 to 48 hr of culture. These cells exhibited a dendritic shape, expressed Ia antigens, lacked Thy-1 antigens, and displayed the ultrastructural features of Langerhans cells (LC) with the notable exception of Birbeck granules. Although after 24 hr, only 20% of Ia+ EC were 3C7/7D4+, the vast majority of LC displayed 3C7/7D4 binding sites after 4 to 5 days of culture. Preincubation of cultured LC-enriched EC with recombinant human IL 2 prevented subsequent 3C7-but not 7D4-binding to these cells. Western blot analysis of 7D4-reactive material of detergent extracts from LC-enriched EC revealed three bands in the same m.w. range as reported for CTLL cells. These results demonstrate that cultured LC express IL 2 receptors and may bear important implications for a better understanding of growth regulation, differentiation, and immunologic functions of LC.
Animals↗
Use of a cocktail of monoclonal antibodies and human complement in selective killing of acute lymphocytic leukemia cells.
Autologous remission bone marrow is a potential source of repopulative stem cells after ablative chemoradiotherapy of tumor patients. Even with remission bone marrow, one major obstacle to use of autologous bone-marrow support is the danger of reinfusing viable tumor cells. This report describes a purging protocol with human complement, which seems suitable for eliminating acute lymphatic leukemia (ALL) cells of the common ALL type. Lysis of ALL blasts is induced with a cocktail of 3 monoclonal antibodies of IgM type (termed VIB-pool). These are directed against the CALLA (CD 10) antigen (VIL-AI antibody) and against 2 different epitopes of the CD24 surface structure (VIB-C5 and VIB-E3 antibodies). The purging efficiency was evaluated with leukemic cell lines of the common ALL type (Reh-6 and Nalm-6) and with blast cells from common ALL patients. Optimal lysis was obtained with antibody and human serum concentrations as low as 1 microgram/ml and 7% respectively. As a standard purging protocol we propose one 20-min incubation at room temperature with antibody followed by two 30-min incubations at 37 degrees C with 25% human complement. In dye exclusion test 99% purging efficiency and in clonogenic assays detecting elimination of up to 5 logs of clonogenic tumor cells 99.99% (= 4 logs) purging efficiency were achieved. Treatment with VIB-pool and human complement had no negative effect on the growth of the normal hemopoietic progenitor cells CFU-GM, CFU-E and BFU-E.
Animals↗
Recombinant interferon-gamma induces interleukin 2 receptors on human peripheral blood monocytes.
The present report describes the inducibility of IL 2 receptors on human peripheral blood monocytes. Although freshly isolated monocytes are IL 2 receptor negative, approximately one-third of the cells react with the anti-Tac antibody within 18 hr of culture. IFN-gamma is found to double both the number of positive cells and the number of binding sites, whereas IL 2 has no influence on the IL 2 receptor expression on monocytes. Anti-Tac precipitates from monocyte lysates several protein bands of similar m.w. to those previously found with activated T and B cells. Finally, IFN-gamma-induced, but not resting, monocytes are found to bind recombinant IL 2. We conclude that IFN-gamma induces peripheral blood monocytes to express IL 2 receptors similar in structure to those found on activated T and B lymphocytes.
Antibodies, Monoclonal↗
T cell stimulation via the erythrocyte receptor. Synergism between monoclonal antibodies and phorbol myristate acetate without changes of free cytoplasmic Ca++ levels.
We observed that certain E-receptor antibodies (CD2 antibodies) can induce proliferation of resting human T cells in the presence of PMA, while other CD2 antibodies fail to have such an effect. The same CD2 antibodies that were mitogenic in the presence of PMA (9.6, X11, VIT13), but not the nonreactive ones, were also able to induce T cell proliferation via the so-called alternative pathway of T cell activation, i.e., when added pairwise in certain combinations to T cells in the absence of PMA. While the simultaneous addition of two comitogenic CD2 antibodies (9.6 or X11 plus VIT13) or the addition of a single nonmitogenic CD3 antibody (VIT3) led to a clearcut elevation of intracellular Ca++ levels, no such effect could be observed after the addition of one CD2 antibody alone. Even in the presence of PMA, one comitogenic CD2 antibody alone was unable to trigger a significant Ca++ response, although this combination induced a proliferative response. These data indicate that, distinguishable by their influence on free cytoplasmic Ca++, there are two different mechanisms of T cell activation via CD2. While simultaneous triggering with two antibodies leads to cell proliferation preceded by an increase of Ca++ levels, stimulation with one antibody plus PMA results in proliferation without a measurable early Ca++ response. We conclude that T cells treated by certain CD2 antibodies alone already recognize an activation signal probably unrelated to Ca++ homeostasis, a signal that can further be developed by PMA to result in a completely developed proliferative response.
Antibodies, Monoclonal↗
Heterogeneity of the first cluster of differentiation: characterization and epitopic mapping of three CD1 molecules on normal human thymus cells.
In humans, the presence of two non-HLA class 1-like molecules, whose expression, similar to murine Tla, is restricted to cortical thymocytes, has been shown with monoclonal antibodies defining the first cluster of differentiation (CD1). We report here with the use of 12 anti-CD1 antibodies and a combination of technical approaches, the characterization of a third CD1 molecule. We show that we can presently define seven different epitopes on the three CD1 molecules: four epitopes are restricted to the 49,000 dalton molecule, two epitopes to the 45,000 dalton molecule, and one epitope to the 43,000 dalton molecule. We show that the association of the newly identified 45,000 dalton heavy chain with human beta2-microglobulin is weak. In addition we show the presence of a fourth non-HLA class I molecular species on the surface of normal human thymus cells.
Animals↗
A rapid and simple immunoperoxidase staining procedure for blood and bone marrow samples.
A fast and simple indirect immunoperoxidase staining technique is described, which can also be used for the characterization of cells with high endogenous peroxidase activity such as many haemopoietic cells. It is based on the combination of a newly developed glucose-oxidase plus glucose procedure for the inhibition of endogenous peroxidase activity with a standard 2 or 3 layer immunoperoxidase staining protocol. Glucose-oxidase plus glucose mixture completely inhibits endogenous peroxidase activity without having a detectable deleterious effect on any of the cellular antigens so far studied. In many instances this permits the use of only 1 layer of horseradish peroxidase (HRP)-labelled antibodies. The glucose-oxidase plus glucose mixture can also be added to cells together with the HRP-labelled antibody solution without losing its inhibitory effect for endogenous peroxidase activity and without leading to a visually detectable loss of the activity of the HRP conjugate. Consequently, the separate incubation step previously necessary for the inhibition of endogenous peroxidase activity becomes superfluous.
Antibodies, Monoclonal↗