摘要:

Plating efficiencies of EBV transformed human B cells seeded in single cell cultures are far lower (<1%) than observed in T cell cloning experiments. This report describes the stimulatory effect of several crude as well as recombinant growth factors on proliferation of EBV transformed B cells measured by [3H]thymidine uptake. Supernatant of LPS activated monocytes (HSF) and recombinant interleukin 4 (rIL-4), but not recombinant IL-1β, IL-2, IL-6, TNFα, GM-CSF, and interferon [unk]increased [3H]thymidine incorporation. The combination of HSF and rIL-4 was found to be synergistic on B cell proliferation. Plating efficiency of EBV transformed B cells at limiting dilution was improved by HSF, but not by the combination of HSF and rIL-

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.1

年代:1990

数据来源: MAL

摘要:

The capacity of adoptively transferred CD8+ effector T cells to induce meningitis in immunosuppressed, or unsuppressed, recipients infected with lymphocytic choriomeningitis virus (LCMV) may be diminished by prior incubation of the lymphocytes with IgM monoclonal antibodies (MAbs) specific for CD8 or Thy1.2. The same is true, though to a lesser extent, for the further proliferation of donor T cells in the spleens of the immunosuppressed mice. This inhibition of cell mediated immunity can be overcome, at least for the unsuppressed recipients, by increasing the numbers of cells that are transferred, even though exposure to MAb+ complement abrogates all cytotoxic T cell activityin vitro.The LCM model thus provides a quantitative system for assessing the consequences of MAb binding for T cell trafficking and effector functionin vivo.

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.9

年代:1990

数据来源: MAL

摘要:

In a recent report we described the identification of physical associations between Major Histocompatibility Complex (MHC) Class II (Ia) antigens and other structures of Mr 67,000, which were significantly enhanced following brief T-B cell co-culture (1). To further investigate this 67K Ia-associated product, monoclonal antibodies (MAb) were produced against isolated 67K material and their reactivity examined. Cell surface binding by these MAb was detected only after perturbation of the membrane by cellular adherence or following aldehyde fixation, which indicates that the determinant recognized by these mAb is retained in the plasma membrane in a covert fashion. All lymphoid cells tested showed reactivity with the MAb as determined by immunofluorescence and by ELISA, but no binding was detected on bone marrow or peritoneal macrophages. Expression of the antigen reactive with these antibodies followed a similar pattern with established murine cell lines, with T and B cell lines and a pre-B cell line showing reactivity, while no antigen was detected on macrophage-like and fibroblast cell lines. The intensity of antigen expression by normal lymphoid cells was ordered: thymocytes>splenic T cells ≥ bone marrow lymphocytes>splenic B cells. No correlation was observed between expression of Ia antigens by non-lymphoid cells and expression of the 67K molecule. These observations suggest that this antigen is primarily a marker of lymphoid cells, with the highest expression on cells of the T lymphocyte lineage. Finally, inhibition of antigen-specific, MHC-restricted T-cell activation by the MAb directed against the 67K structure suggests an important functional role for this interesting molecule originally identified by its physical association with Ia following T-B cell interaction

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.17

年代:1990

数据来源: MAL

摘要:

A monoclonal antibody was made after immunization of mice with the 1-32 amino terminal peptide of the alpha subunit of 32K human ovarian inhibin. The IgG2a mouse antibody reacted 6 times better with bovine 1-32 peptide than it did with 32K bovine inhibin. By contrast sheep polyclonal antibodies made by a similar method had a 29 fold bias in reactivity towards the immunizing peptide. Relative to homologous 1-32 peptide standards, the monoclonal antibody measured apparently higher amounts of immunoreactive material(s) in human (13.5 fold) and bovine (27 fold) follicular fluids than did the polyclonal anti 1-32 peptide antibodies. Immunochemical studies revealed that the epitope recognized by the monoclonal antibody was different from the major epitope recognized by the polyclonal antibodies. The monoclonal antibody reacted much better with human inhibin 1-32 sequences than with bovine (73 fold) or porcine (23 fold). Although the 32K form of human inhibin has not yet been purified, it can be inferred that the monoclonal antibody would be able to detect as little as 2 ng/ml of 32K human inhibin in competitive radioimmunoassays. The antibody must also react with some of the multiple molecular forms of inhibin found in human follicular fluids, and it was shown to function well in the quantitative immunoaffinity extraction of inhibin-like immunoreactivity from follicular fluid. It seems likely that this monoclonal antibody will prove a useful tool for research on human inhibin.

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.31

年代:1990

数据来源: MAL

摘要:

Mouse-human chimeric antibody specific for human carcinoembryonic antigen (CEA) was produced by recombinant DNA techniques. The genes of the mouse variable regions of heavy and light chains were cloned from the mouse hybridoma, 2.7.1G.10., which secreted anti human CEA antibody (IgG1,κ), and were joined with human γ1 and κ constant genes. The affinity of the resultant chimeric antibody to its relevant antigen was the same as that of the parental mouse monoclonal antibody when analysed by Scatchard plot analysis. The chimeric antibody showed a potent antibody dependent cell-mediated cytotoxicity (ADCC) activity with human peripheral blood mononuclear cells against CEA-positive human adenocarcinoma cells. In vivo imaging analysis revealed that the present chimeric antibody was specifically localized on the tumor site. These results indicate that our mouse-human chimeric antibody is a promising reagent for the diagnosis and therapy of CEA-positive human cance

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.43

年代:1990

数据来源: MAL

摘要:

Eleven monoclonal antibodies against chicken gizzard talin, a major focal adhesion protein, have been produced. In order to determine the degree of homology between talin molecules from different sources, these antibodies were used to immunolocalize talin in five different vertebrate species. Three representative talin antibodies are described in this report.

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.57

年代:1990

数据来源: MAL

摘要:

We established 16 monoclonal antibodies (MAbs) to endothelin (ET), a novel and potent vasoconstrictor. They were classified into two groups: eight of them reacted with the C-terminal portion of ET, and the other eight reacted with the non-C-terminal portion. We constructed the sandwich enzyme linked immunosorbent assays (ELISAs) using a combination of a MAb belonging to the former with that belonging to the latter, and could detect about 400 pg/ml of ET.

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.63

年代:1990

数据来源: MAL

摘要:

We have produced and characterized the first monoclonal antibody against neopterin (D-erythro-6-(1,2,3,-trihydroxypropyl)pterin). The antibody specifically recognizes neopterin in a modified RIA. The binding capacity in this assay is 34%, the sensitivity limit of inhibition is 0.9 nmol/1. Cross-reactivity exists with monapterin (L-threo(1,2,3,trihydroxypropyl)pterin) in 30%, with other pteridines cross-reactivity has been found in less than 5%.

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.71

年代:1990

数据来源: MAL

摘要:

A method for thein vitroimmunization of human tonsilar lymphocytes is described. This method allows for the senitization of B-cells against various antigens under serum-free medium conditions. The medium is supplemented with defined concentrations of antigen, rIL-2 and muramyl dipeptide to boost the immune responses. This method may prove useful for the generation of antigen reactive B-cells for use in human monoclonal antibody production and the study of the effects of other lymphokines on thein vitroimmune response.

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.81

年代:1990

数据来源: MAL

摘要:

The use of immunoperoxidase staining methods to detect human monoclonal antibodies that react with frozen tissue sections of human tumors is limited due to endogenous immunoglobulin present in the sections. The endogenous immunoglobulin produces a significant background staining which makes the detection of a specific reaction difficult and unreliable. A method of incubating the sections with papain fragments (Fab') of goat anti-human immunoglobulin was evaluated. It was demonstrated that Fab' fragments of goat anti-human polyvalent immunoglobulin can block endogenous immunoglobulin background staining after prolonged washing of tissue sections (20h) and thus facilitating the screening of monoclonal antibodies against frozen sections of tumor tissue.

ISSN:0272-457X    

DOI:10.1089/hyb.1990.9.91

年代:1990

数据来源: MAL