==================================BSR23================================== 23. Generation of monoclonal antibodies against hormones and T cell soluble factors and their receptors. - Method for the in vitro immunization - Method for the immunization of very small amounts of antigens (gel. slice, etc.). 1 UI - 87103733 AU - Volk HD ; Brocke S ; Osawa H ; Diamantstein T TI - Effects of in-vivo administration of a monoclonal antibody specific for the interleukin-2 receptor on the acute graft-versus-host reaction in mice. AB - Parental strain T lymphocyte injected into F1 mice respond to allogeneic MHC antigens and so induce the symptoms of a graft-versus-host reaction (GVHR). We have measured the local GVHR by the popliteal lymph node assay, and showed the suppression of the local GVHR in mice by treatment with the monoclonal antibody (MoAb) AMT-13 which is specific against the interleukin 2 (IL-2) receptor on activated mouse lymphocytes. The inhibitory effect of the AMT-13 administration was comparable with the suppression of the local GVHR by treatment with L3T4, an MoAb directed against the T helper subset. The L3T4 administration caused a dramatic decrease in the proportion of the cells with the L3T4 phenotype in the circulation and a marginal reduction of these cells in the lymph nodes. In contrast, the AMT-13 treated mice showed no changes in the distribution of the T lymphocyte subsets besides those in the GVHR-stimulated lymph nodes. Obviously, only the small subset of antigen-activated IL-2 receptor-bearing lymphocytes was influenced by treatment with AMT-13. MoAb directed against antigens whose expression is restricted to activated lymphocytes, such as the IL-2 receptor, might become useful for a short term immunosuppression with limited side effects. MH - Animal ; Antibodies, Monoclonal/*THERAPEUTIC USE ; Antibody Specificity ; Female ; *Graft vs Host Reaction ; Immunosuppression/*METHODS ; Lymph Nodes/IMMUNOLOGY ; Male ; Mice ; Mice, Inbred DBA ; Receptors, Antigen, T-Cell/IMMUNOLOGY ; Receptors, Immunologic/*IMMUNOLOGY ; Support, Non-U.S. Gov't ; T Lymphocytes/CLASSIFICATION/IMMUNOLOGY SO - Clin Exp Immunol 1986 Oct;66(1):126-31 2 UI - 87058993 AU - Leo O ; Sachs DH ; Samelson LE ; Foo M ; Quinones R ; Gress R ; Bluestone JA TI - Identification of monoclonal antibodies specific for the T cell receptor complex by Fc receptor-mediated CTL lysis. AB - Monoclonal antibodies (mAb) directed at the T cell receptor complex (TcR) on cloned T cells have generally been identified by their ability to inhibit the clone's antigen-specific function. Because such inhibition is highly dependent on antibody concentration and affinity, detection of anti-clonotypic antibodies to murine alloreactive T cells has been very difficult. In this report, an alternative method is described on the basis of the ability of antibodies specific for the TcR complex to activate T cells in an antigen-independent manner. The assay is based upon the observation that soluble antibodies to human T3 promote lysis of irrelevant, Fc receptor-positive targets by a human CTL line. By using this approach, an anti-TcR mAb has been identified among a panel of murine mAb generated against an alloreactive CTL clone. Induction of lysis by soluble anti-TcR mAb has been shown to require both the expression of Fc receptors on the target cell and conjugate formation between the effector and the target cell. This assay provides a screening procedure that is much more sensitive than inhibition of function, and it preferentially detects antibodies specific for cell surface molecules involved in T cell activation. MH - Animal ; Antibodies, Monoclonal/*IMMUNOLOGY/ISOLATION & PURIFICATION ; Antigens, Surface/*IMMUNOLOGY ; Cell Communication ; Cell Line ; Cytotoxicity, Immunologic ; Lymphocyte Transformation ; Mice ; Receptors, Antigen, T-Cell/*IMMUNOLOGY ; Receptors, Fc/*IMMUNOLOGY ; T Lymphocytes, Cytotoxic/*IMMUNOLOGY SO - J Immunol 1986 Dec 15;137(12):3874-80 3 UI - 87041525 AU - McDuffie M ; Born W ; Marrack P ; Kappler J TI - The role of the T-cell receptor in thymocyte maturation: effects in vivo of anti-receptor antibody. AB - The T-cell receptor, which recognizes antigen plus a product of the major histocompatibility complex, has been postulated to drive T-cell maturation in the thymus by engaging major histocompatibility complex proteins expressed on thymic stromal cells. We tested this idea by injecting neonatal animals with an anti-receptor antibody, KJ16, that binds to about 20% of T cells and is capable of blocking receptor function. In the presence of this antibody, mature T cells bearing the KJ16 epitope failed to develop. On the other hand, although the antibody could be shown to bind to receptors on cortical thymocytes, it did not prevent the rapid expansion or survival of the bulk of the KJ16+ cells in this population. These results are consistent with the hypothesis that most cortical thymocytes arise by a receptor-independent mechanism and that only a small proportion of these cells mature by a process dependent on receptor-major histocompatibility complex interactions. MH - Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Cell Differentiation ; Immunoglobulins, Fab/IMMUNOLOGY ; Kinetics ; Major Histocompatibility Complex ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Pepsin/ PHARMACODYNAMICS ; Receptors, Antigen, T-Cell/IMMUNOLOGY/*PHYSIOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; T Lymphocytes/ *PHYSIOLOGY SO - Proc Natl Acad Sci USA 1986 Nov;83(22):8728-32 4 UI - 87032078 AU - Wang CY ; Bushkin Y ; Pica R ; Lane C ; McGrath H ; Posnett DN TI - Stimulation and expansion of a human T-cell subpopulation by a monoclonal antibody to T-cell receptor molecule. AB - A murine monoclonal antibody (MAb) was obtained that showed unique specificity for the immunizing T-cell line HPB-ALL. This antibody, C37 (an IgG1,K) also reacted with a small (2-5%) population of normal peripheral blood T (PBL-T) cells. These C37-positive (C37+) cells were found in both the T4/Leu3+ and T8/Leu2+ subsets. Like OKT3 antibody, C37 induced T-cell mitogenesis with a peak proliferative response at day 3. In long-term cultures containing irradiated autologous feeder cells and IL-2, C37 antibody caused the selective expansion of C37+ T cells. On HPB-ALL cells C37 induced comodulation of the T3 molecule. C37 precipitated a disulfide-linked dimer characteristic of the T-cell antigen receptor consisting of an alpha-subunit (45-48 kD) and a beta-subunit (38-42 kD) from both C37+ T-cell blasts of a normal individual and HPB-ALL cells that were surface radioiodinated. However, the precipitated molecule isolated from C37 antibody-activated T-cell blasts exhibited a different pI from that isolated from HPB-ALL cells. Our studies indicate that C37 recognizes an epitope on the T-cell receptor molecule that is shared by a subpopulation of human T cells, which raises the possibility that multiple variable-region associated and/or framework-like determinants of the T-cell antigen receptor can be defined serologically and used in functional and molecular studies of T-cell subsets. MH - Antibodies, Monoclonal/*IMMUNOLOGY ; Antigenic Determinants/IMMUNOLOGY ; Cell Line ; Human ; Leukemia, Lymphoblastic ; Lymphocyte Transformation ; Receptors, Antigen, T-Cell/*IMMUNOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; T Lymphocytes/*CLASSIFICATION/IMMUNOLOGY SO - Hybridoma 1986 Fall;5(3):179-90 5 UI - 87017051 AU - Posnett DN ; Wang CY ; Friedman SM TI - Inherited polymorphism of the human T-cell antigen receptor detected by a monoclonal antibody. AB - Three different murine monoclonal antibodies to the human clonotypic T-cell antigen receptor immunoprecipitate the alpha-beta chain heterodimer; induce comodulation of the clonotypic molecule with the T3 molecular complex; stain small populations of normal polyclonal T cells, suggesting that they react with variable or joining region determinants of the clonotypic receptor; and induce proliferation of resting T cells. While two of these antibodies detect the clonotypic receptor in all individuals studied, the third antibody (OT145), described herein, does not detect the T-cell antigen receptor on T cells of all individuals. By indirect immunofluorescence, three groups can be distinguished within a population of individuals (n = 138) by OT145. Individuals lacking T cells reactive with OT145 have a homozygous OT145-phenotype. T cells from such individuals fail to proliferate in the presence of OT145 in contrast to T cells from OT145+ individuals. Individuals with a relatively large percentage of OT145+ T cells, 4.5 +/- 1.54% (mean +/- 2 SEM) are homozygous OT145+, while those with an intermediate percentage, 2.04 +/- 0.9%, have a heterozygous phenotype. Family studies suggest autosomal codominant inheritance of the OT145 phenotype. The distribution of the three OT145-defined phenotypes varies considerably in populations of different ethnic background. Taken together these data suggest that the polymorphism detected by OT145 may represent a variable or joining region allotypic system of the human T-cell antigen receptor. In addition, our results indicate that allelic exclusion governs the expression of the clonotypic receptor by human T cells. MH - Alleles ; Antibodies, Monoclonal/*IMMUNOLOGY ; Ethnic Groups ; Human ; Immunoglobulin Allotypes/ANALYSIS ; *Polymorphism (Genetics) ; Receptors, Antigen, T-Cell/*ANALYSIS/IMMUNOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Proc Natl Acad Sci USA 1986 Oct;83(20):7888-92 6 UI - 87013626 AU - Clark DM ; Boylston AW ; Hall PA ; Carrel S TI - Antibodies to T cell antigen receptor beta chain families detect monoclonal T cell proliferation [published erratum appears in Lancet 1986 Dec 6;2(8519):1342] AB - The T cell antigen receptor is constructed from independent gene segments much like those used to assemble immunoglobulin genes. One of the receptor's two protein subunits, the beta chain, uses a limited number of variable region segments. The product of these V region segments can be identified by monoclonal antibodies and can be used to define populations of normal T cells which use the same V beta (V beta) gene segment. These antibodies have been used to define monoclonality or its absence in T cell populations. Twenty-four cases have been studied (twenty with solid T cell lymphomas and four with T cell leukaemias). Two monoclonal antibodies to V beta (anti-HPB-A11 and anti-Jurkat) were tested, and three cases of T cell lymphoma were positive, two to anti-Jurkat and one to anti-HPB. The malignant nature of T cell proliferations can be directly diagnosed in tissue sections and intact cell suspensions. This approach should also make possible the monitoring of changes in malignant populations in response to therapy. MH - Antibodies, Monoclonal/*DIAGNOSTIC USE ; Human ; Immunoglobulin Variable Region/*IMMUNOLOGY ; Leukemia/DIAGNOSIS/IMMUNOLOGY ; *Lymphocyte Transformation ; Lymphoma/DIAGNOSIS/IMMUNOLOGY ; Receptors, Antigen, T-Cell/*IMMUNOLOGY ; Stains and Staining ; Support, Non-U.S. Gov't ; T Lymphocytes/*IMMUNOLOGY SO - Lancet 1986 Oct 11;2(8511):835-7 7 UI - 87009874 AU - Lanier LL ; Ruitenberg JJ ; Allison JP ; Weiss A TI - Distinct epitopes on the T cell antigen receptor of HPB-ALL tumor cells identified by monoclonal antibodies. AB - The T cell antigen receptor is a approximately 90,000 dalton disulfide linked heterodimer that is non-covalently associated with the CD3 complex. Prior studies have demonstrated that anti-CD3 or -Ti antibodies can mimic antigen and induce cellular proliferation and the secretion of lymphokines. An early event in activation via CD3/Ti is a rapid increase in concentration of intracellular Ca2+ levels. In the present studies, we have produced a panel of monoclonal antibodies (MAb) against the Ti expressed on HPB-ALL tumor cells. All MAb immunoprecipitate a approximately 90,000 dalton disulfide linked heterodimer and induced co-modulation of Ti and CD3. On the basis of competitive binding studies, four distinct epitopes on the Ti of HPB-ALL were identified with MAb L38, L39, L41, and L42. These epitopes were additionally discriminated on the basis of reactivity with normal polyclonal T cell populations and functional effects on HPB-ALL. L39 reacted with a monomorphic epitope present on approximately 2 to 5% of peripheral blood T lymphocytes from all donors examined and was specifically mitogenic for peripheral blood T cells expressing this epitope. L39+ T cells in blood included both CD4+ and CD8+ lymphocytes. In contrast, L38, L41, and L42 failed to react with peripheral blood T cells and were not mitogenic for peripheral blood lymphocytes. Anti-Leu-4, L38, L39, and L41 MAb all induced a rapid increase in (Ca2+)i in HPB-ALL tumor cells, similar to previous findings with anti-CD3 and anti-Ti MAb against various tumor cells and peripheral blood T cells. In contrast, L42 MAb did not induce a substantial increase in (Ca2+)i. Failure of L42 to induce a substantial increased (Ca2+)i could not be attributed to the apparent titer, avidity, or isotype of the antibody. These findings suggest that induction of increased (Ca2+)i upon binding of Ti is epitope dependent. Furthermore, these data demonstrate that several distinct public and private epitopes can be identified on the T cell antigen receptor. MH - Antibodies, Monoclonal/*IMMUNOLOGY ; Antibody Specificity ; Antigenic Determinants ; Antigens, Surface/IMMUNOLOGY ; Binding, Competitive ; Calcium/PHYSIOLOGY ; Cell Line ; Flow Cytometry ; Human ; Immunologic Capping ; Lymphocyte Transformation ; Receptors, Antigen, T-Cell/ *IMMUNOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Immunol 1986 Oct 1;137(7):2286-92 8 UI - 86247815 AU - Carrel S ; Giuffr:e L ; Vacca A ; Salvi S ; Mach JP ; Isler P TI - Monoclonal antibodies against idiotypic determinant(s) of the T cell receptor from HPB-ALL cells induce IL2 production in Jurkat cells without apparent evidence of binding. AB - Two monoclonal antibodies (mAb) directed against idiotypic determinants of the T cell receptor (anti-Ti) from HPB-ALL cells induce interleukin 2 (IL2) production in Jurkat T cells without evidence of binding to these cells as judged by fluorescence-activated cell sorter (FACS) analysis, indirect antibody-binding radioimmunoassay and direct binding studies with 125I-labeled mAb. The IL2 response induced by these mAb observed both in the presence and absence of phorbol myristate acetate was in the range of that obtained when Jurkat cells were stimulated with phytohemagglutinin or anti-T3 mAb (Leu 4). The idiotypic specificity of the two anti-HPB-ALL Ti mAb was demonstrated by several criteria. Both mAb bound specifically to HPB-ALL cells as determined by radioimmunoassay or FACS analysis but not with 8 other T cell lines. The anti-HPB-ALL Ti mAb precipitated a disulfide-linked heterodimer of 85 kDa only from 125I-labeled HPB-ALL cells and not from other cell lines tested. Incubation of HPB-ALL cells with anti-T3 abrogated the expression of T3 and induced co-modulation of the idiotypic structures detected by the two anti-HPB-ALL Ti mAb. Conversely, incubation of HPB-ALL cells with either one of the anti-Ti mAb abrogated the expression of T3 and of the idiotypic structures. Our results suggest that mAb with an apparent unique specificity for the receptor of the immunizing T cell line HPB-ALL can activate Jurkat cells by a very weak cross-reaction with these cells, which is not detectable by conventional binding tests. MH - Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antigen-Antibody Reactions ; Cell Separation ; Comparative Study ; Cross Reactions ; Flow Cytometry ; Human ; Immunoglobulin Idiotypes/IMMUNOLOGY ; Immunologic Technics ; Interleukin 2/*BIOSYNTHESIS ; Leukemia, Lymphoblastic/*IMMUNOLOGY ; *Lymphocyte Transformation ; Mice ; Receptors, Antigen, T-Cell/ *IMMUNOLOGY ; T Lymphocytes/*IMMUNOLOGY/METABOLISM SO - Eur J Immunol 1986 Jul;16(7):823-8 9 UI - 86298417 AU - Jondal M ; Kullman C ; Alter MB ; Ljunggren K TI - Monoclonal antibodies against the NK cell-FcR and the T3-complex potentiate normal lymphocyte killing. AB - Pretreatment of normal human lymphocytes with monoclonal IgG against the NK cell-FcR (IgG) or the T3 complex was found to potentiate killing of most NK sensitive target cells with the exception of T-cell derived cells. Anti-FcR IgM monoclonals were suppressive for all target cells. IgG anti-FcR mediated potentiation required minute amounts of antibody but was also seen at high anti-FcR concentrations that modulated FcR activity. Potentiated and FcR modulated cells retained anti-FcR IgG on the membrane and conjugated normally to target cells. Anti-FcR potentiation blocked antibody-dependent killing but did not influence lectin-dependent killing, with anti-T3 the opposed effect was seen. Combined anti-FcR and anti-T3 treatment resulted in decreased potentiation. The results suggest that the NK cell-FcR may be activated during normal NK cell killing (without the addition of antibody) as suggested for FcR in B cell triggering. MH - Antibodies, Monoclonal/*IMMUNOLOGY ; Antibody-Dependent Cell Cytotoxicity ; Antigens, Surface/*IMMUNOLOGY ; Cells, Cultured ; *Cytotoxicity, Immunologic ; Human ; Immunity, Cellular ; In Vitro ; Killer Cells, Natural/*IMMUNOLOGY ; Lectins/IMMUNOLOGY ; Receptors, Antigen, T-Cell/ IMMUNOLOGY ; Receptors, Fc/*IMMUNOLOGY ; Rosette Formation ; Support, Non-U.S. Gov't ; T Lymphocytes/*IMMUNOLOGY SO - Cell Immunol 1986 Jun;100(1):158-66 10 UI - 86252246 AU - Boylston AW ; Borst J ; Yssel H ; Blanchard D ; Spits H ; de Vries JE TI - Properties of a panel of monoclonal antibodies which react with the human T cell antigen receptor on the leukemic line HPB-ALL and a subset of normal peripheral blood T lymphocytes. AB - Five Mab raised against the T cell antigen receptor of the human T cell line HPB-ALL which react with a subpopulation of normal peripheral blood T cells are described. Three Mab, 3D6, 1C1, and 1C2, react with 3 to 5% of normal PBL and stimulate proliferation of the cells with which they react. An increase in the number of cells which react with all five Mab occurs. Two Mab, 2D4 and 65, react with subsets of the cells which bind 1C1, 1C2, and 3D6 and divide the family into four subgroups, 2D4+ 65+, 2D4+ 65-, 2D4- 65+, and 2D4- 65-. Functional T cell clones in all four subfamilies have been observed. Cytolytic function can be correlated with the TcR phenotype expressed because all of the Mab which react with a particular clone inhibit its ability to lyse a specific target. The epitopes recognized by the panel are closely related because all five block each other's binding to HPB-ALL. In addition, the determinants recognized by 3D6, 1C1, and 1C2 on normal lymphocytes are probably very closely related because all clones examined react with all three Mab. MH - Animal ; Antibodies, Monoclonal/*ANALYSIS/ISOLATION & PURIFICATION/ PHYSIOLOGY ; Antigen-Antibody Reactions ; Antigenic Determinants/ IMMUNOLOGY ; Binding, Competitive ; Cell Line ; Cytotoxicity, Immunologic ; Female ; Human ; Leukemia, Lymphoblastic/*IMMUNOLOGY ; Lymphocyte Transformation ; Mice ; Mice, Inbred BALB C ; Phenotype ; Receptors, Antigen, T-Cell/*IMMUNOLOGY ; Support, Non-U.S. Gov't ; T Lymphocytes/ *CLASSIFICATION/IMMUNOLOGY/METABOLISM SO - J Immunol 1986 Jul 15;137(2):741-4 11 UI - 86164584 AU - Staerz UD ; Bevan MJ TI - Activation of resting T lymphocytes by a monoclonal antibody directed against an allotypic determinant on the T cell receptor. AB - The murine monoclonal antibody F23.1 reacts with an allotypic determinant on the beta chain of the T cell receptor expressed by approximately 20% of T helper and cytotoxic T lymphocytes of most common mouse strains. This IgG2a antibody, either in soluble form or covalently coupled to Sepharose beads, can activate resting T cells from naive animals to proliferate. Interestingly, under all conditions of activation, the antibody can only induce proliferation if exogenous lymphokines in the form of Con A supernatant are provided. Thus, it is unlike most lectins and anti-T3 antibodies in this regard. Furthermore, under all conditions of culture, F23.1 activates preferentially the Lyt-2+ subset of T cells. This is the case even in the presence of accessory cells. Further evidence is provided that two soluble lymphokines, different from IL2, are required to initiate IL2-dependent growth and to allow the expression of lytic activity. MH - Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antigenic Determinants/ IMMUNOLOGY ; Antigens, Ly/IMMUNOLOGY ; Cells, Cultured ; Cytotoxicity, Immunologic ; Interleukin 2/PHARMACODYNAMICS ; *Lymphocyte Transformation/ DRUG EFFECTS ; Lymphokines/PHARMACODYNAMICS ; Mice ; Mice, Inbred BALB C ; Rats ; Rats, Inbred LEW ; Receptors, Antigen, T-Cell/*IMMUNOLOGY ; Support, U.S. Gov't, P.H.S. ; T Lymphocytes/*IMMUNOLOGY SO - Eur J Immunol 1986 Mar;16(3):263-70 12 UI - 86113387 AU - Tadmori W ; Kant JA ; Kamoun M TI - Down regulation of IL 2 mRNA by antibody to the 50-kd protein associated with E receptors on human T lymphocyte. AB - Recent studies have shown that antibodies to certain epitopes on the 50-kd molecule associated with sheep erythrocyte receptors on human T cells can suppress T cell proliferation and interleukin 2 (IL 2) elaboration. We used a human IL 2 cDNA clone to investigate the effect of antibody 9.6 and cyclosporin A (CsA) on the regulation of IL 2 mRNA levels in the cloned human leukemic T cell line Jurkat, J32. Maximal levels of IL 2 mRNA were reached 6 hr after induction of Jurkat cells with a combination of mitogen phytohemagglutinin (PHA) and phorbol ester (TPA). Antibody 9.6, added during the first 4 hr after lymphocyte stimulation, markedly inhibited IL 2 mRNA accumulation induced by low but synergistic combination of PHA (5 micrograms/ml) and TPA (1.0 ng/ml). The inhibition by 9.6 was not demonstrable as the concentration of PHA or TPA was increased. In contrast, the ability of CsA to suppress IL 2 mRNA accumulation appeared to be independent of PHA or TPA concentration and was minimal if CsA was added 4 hr after stimulation. IL 2 mRNA could be superinduced several folds by addition of cycloheximide 3 hr after induction of J32 with mitogens. Antibody 9.6 did not prevent IL 2 mRNA superinduction induced by cycloheximide, whereas CsA, as well as transcription inhibitor DRB, completely blocked this phenomenon. These findings indicate that signals induced by antibody 9.6 regulate IL 2 production at a pre-translational level, are operative for an extended period of time overlapping with the early phase of IL 2 mRNA accumulation, suppress IL 2 gene expression induced by PHA as well as TPA, and that antibody 9.6 and CsA exert their inhibitory effect by distinct mechanism(s). MH - Animal ; Antibodies, Monoclonal/*PHYSIOLOGY ; Blood Groups/*IMMUNOLOGY ; Cell Line ; Cycloheximide/PHARMACODYNAMICS ; Cyclosporins/ PHARMACODYNAMICS ; Dose-Response Relationship, Immunologic ; Human ; Immune Tolerance ; Interleukin 2/*METABOLISM ; Kinetics ; Lymphocyte Transformation/DRUG EFFECTS ; Molecular Weight ; Phytohemagglutinins/ PHARMACODYNAMICS ; Receptors, Antigen, T-Cell/*METABOLISM ; RNA, Messenger/*BIOSYNTHESIS ; Sheep ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; T Lymphocytes/IMMUNOLOGY/*METABOLISM ; Tetradecanoylphorbol Acetate/PHARMACODYNAMICS SO - J Immunol 1986 Feb 15;136(4):1155-60 13 UI - 86149322 AU - Staerz UD ; Bevan MJ TI - Hybrid hybridoma producing a bispecific monoclonal antibody that can focus effector T-cell activity. AB - Previous studies have shown that heteroconjugates of monoclonal antibodies in which one of the component antibodies is directed at the T-cell receptor and the other is directed against any chosen site can focus effector T cells to function at the targeted site. We report here the production of a hybrid hybridoma cell line, H1.10.1.6, which secretes large amounts of a bispecific hybrid antibody of the IgG2a class, that can focus T-cell activity. The parental hybridoma lines for the secondary fusion were F23.1, which secretes an antibody specific for an allotypic determinant on the T-cell receptor of most mouse strains, and 19E12, secreting an anti-Thy-1.1 antibody. The bispecific hybrid antibody was partially purified by hydroxylapatite chromatography and characterized by isoelectric focusing. It efficiently targets Thy-1.1-expressing tumor cells for lysis by F23.1 receptor-positive cytotoxic T-cell clones in vitro. Such hybrid antibodies produced by hybrid hybridoma cell lines may have application in the therapeutic targeting of tumors or sites of viral infections for attack by T cells. MH - Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antibody Specificity ; Antigens, Surface/*IMMUNOLOGY ; Cytotoxicity, Immunologic ; Hybridization ; Immunity, Cellular ; Immunoglobulins, Fab/IMMUNOLOGY ; Mice ; Receptors, Antigen, T-Cell/*IMMUNOLOGY ; Support, U.S. Gov't, P.H.S. ; T Lymphocytes, Cytotoxic/*IMMUNOLOGY SO - Proc Natl Acad Sci USA 1986 Mar;83(5):1453-7 14 UI - 86113416 AU - Chan PY ; Takei F TI - Expression of a T cell receptor-like molecule on normal and malignant murine T cells detected by rat monoclonal antibodies to nonclonotypic determinants. AB - A T cell receptor-like molecule with a dimer structure of 45 kilodaltons (Kd) under reducing and 90 Kd under nonreducing conditions was detected on the surface of two murine T lymphoma lines, EL-4 and MBL-2, by two rat monoclonal antibodies. The two antibodies seemed to react with different determinants on the same molecule. The antibodies did not react with the surface of normal T cells as tested by flow cytometric analysis of cell surface staining. Two-dimensional gel electrophoresis (IEF vs SDS-PAGE) and tryptic peptide analysis revealed the molecule to consist of two chains with different isoelectric points and different tryptic peptides. A conventional antiserum was raised against the heterodimer purified from EL-4 cells. The immune serum did not bind to the surface of normal T cells. However, the immune serum as well as the monoclonal antibodies immunoprecipitated the dimer molecules from detergent-solubilized normal thymocytes and spleen cells. The dimer molecule was detected on both immature and mature thymocytes. These results suggest that the antibodies detect non-clonotypic determinants on a T cell receptor-like protein. The determinants are masked on the surface of normal T cells, whereas they are exposed on the surface of at least two T lymphoma cell lines. Three polypeptides of 30 Kd, 25 Kd, and 15 Kd were also coprecipitated with the heterodimer from MBL-2 cells. These proteins may associate with the heterodimer and may be masking the antigenic determinants on normal T cells. The relationship between the heterodimer molecule described here and the T cell antigen receptor or the human T cell antigen 9.3 is still unknown. MH - Animal ; *Antibodies, Monoclonal ; Antigen-Antibody Reactions ; Antigenic Determinants/*IMMUNOLOGY/ISOLATION & PURIFICATION ; Antigens, Neoplasm/ ANALYSIS/IMMUNOLOGY/ISOLATION & PURIFICATION ; Antigens, Surface/ANALYSIS/ IMMUNOLOGY/ISOLATION & PURIFICATION ; Binding Sites, Antibody ; Cell Line ; Clone Cells/METABOLISM ; Immune Sera ; Lymphoma/IMMUNOLOGY/*METABOLISM ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Precipitin Tests ; Rats ; Rats, Inbred F344 ; Receptors, Antigen, T-Cell/*ANALYSIS/ IMMUNOLOGY/ISOLATION & PURIFICATION ; Support, Non-U.S. Gov't ; T Lymphocytes/IMMUNOLOGY/*METABOLISM SO - J Immunol 1986 Feb 15;136(4):1346-53 15 UI - 86087177 AU - Borst J ; Boylston AW ; de Vries JE ; Spits H TI - Human T cell lines differing in phenotype and specificity are reactive with the same anti-idiotypic antibody. AB - 3D6, a monoclonal antibody selected for reactivity with the T cell antigen receptor on the T leukemic cell line HPB-ALL, was found to react with 3 to 13% of peripheral blood T lymphocytes of 10 out of 15 normal donors. Peripheral T cells of two donors were stimulated with allogeneic cells, and the 3D6+ cells were enriched by rosetting 3D6-coated cells with goat anti-mouse-coupled human red blood cells and were expanded in interleukin 2-containing medium. In this way, 90 to 100% 3D6+ cell lines were obtained that were cytotoxic for the allogeneic stimulator cells. 3D6 antibody could block antigen-specific cytotoxicity, as well as induce nonspecific cytotoxicity toward target cells that could not be killed in the absence of the 3D6 antibody. The 3D6+ cell populations contained T4+, as well as T8+ cells, indicating that 3D6 antibody defined a T cell receptor population that might harbor various antigenic specificities. One 3D6+ cell line was separated into T4+ T8- and T4- T8+ populations. 3D6 reactive T cell receptors isolated from HPB-ALL and normal cell lines were analyzed biochemically by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, and V8 protease peptide mapping. Isoelectric focusing analysis provided additional evidence for the idea that 3D6 antibody detected a number of structurally distinct T cell receptors, because the T cell receptor alpha-chain was homogeneous in charge after desialation on the clonal tumor line HPB-ALL, but remained heterogeneous in charge on the 3D6+ normal cell lines. No great differences in charge were found between T cell receptors isolated from T4+ and T8+ 3D6+ lines, but their isoelectric focusing patterns were not identical. V8 protease peptide mapping revealed structural differences between the T cell receptor alpha-chain isolated from HPB-ALL on one hand and from the normal 3D6+ lines on the other, whereas the beta-chains did not differ greatly in primary structure according to this analysis. In addition, the peptide mapping suggested differences in primary structure between T cell receptors present on the T4+ population vs those present on the T8+ populations. MH - Anti-Antibodies/PHYSIOLOGY ; Antibodies, Monoclonal/*PHYSIOLOGY ; Antigenic Determinants/ANALYSIS/*IMMUNOLOGY ; Antigens, Neoplasm/ANALYSIS ; B Lymphocytes/IMMUNOLOGY ; Cell Line ; Cell Separation ; Cell Transformation, Viral ; Cross Reactions ; Cytotoxicity, Immunologic ; Human ; Immunoglobulin Idiotypes/*IMMUNOLOGY ; Leukemia, Lymphoblastic/ IMMUNOLOGY ; Lymphocyte Transformation ; Phenotype ; Receptors, Antigen, T-Cell/ANALYSIS ; Rosette Formation ; Support, Non-U.S. Gov't ; T Lymphocytes/CLASSIFICATION/*IMMUNOLOGY/METABOLISM SO - J Immunol 1986 Jan;136(2):601-8