Monoclonal antibodies that inhibit activation and proliferation of lymphocytes. I. Expression of the antigen on monocytes and activated lymphocytes.

It has been shown previously that HBJ127 and HBJ98 monoclonal antibodies raised against a human bladder cancer cell line, and B3 monoclonal antibody against a rat bladder cancer cell line recognized unique cell surface antigens abundant in proliferating cells of the corresponding species. Distribution of the antigens and kinetics of the appearance on human and rat lymphoid cells were examined by means of flow cytometry. Rat macrophages and human peripheral blood monocytes were stained strongly with the B3 and HBJ127 monoclonal antibodies, respectively.

Inhibition of tumor cell growth in vitro by murine monoclonal antibodies that recognize a proliferation-associated cell surface antigen system in rats and humans.

The mouse monoclonal antibody (MoAb) B3 raised against a rat bladder cancer cell line and the MoAbs HBJ127 and HBJ98 raised against a human bladder cancer cell line recognize homologous antigens predominantly present on proliferating cells of the corresponding species. Examination of MoAb-defined antigen and epitopes revealed that both HBJ127 and HBJ98 MoAbs defined a human cell surface glycoprotein complex having an apparent molecular weight of 125,000-130,000 which was composed of a heavy subunit of a glycoprotein nature (Mr 90,000-95,000) and a disulfide-linked light subunit of protein nature (Mr 30,000-35,000), but the HBJ127 and HBJ98 MoAbs recognized a protein epitope and a sugar epitope on the heavy subunit, respectively. Likewise, the B3 MoAb recognized a protein epitope on the heavy subunit of a rat cellular glycoprotein complex of similar composition to the HBJ127/HBJ98-defined human antigen.

3-Methoxy-4-aminoazobenzene, a selective inducer for a high spin form of cytochrome P-448 in rat liver microsomes.

Treatment of Sprague Dawley rats with 3-methoxy-4-aminoazobenzene (3-MeO-AAB) resulted in striking increase of the activity of hepatic microsomal cytochrome P-450s which could efficiently catalyze the mutagenic activation of hepatocarcinogenic aromatic amines such as a tryptophan-pyrolysate component, Trp P-2, and a glutamic acid-pyrolysate component, Glu P-1. The 3-MeO-AAB-induced cytochrome P-450 (3-MeO-AAB-P-450) was examined for the molecular character by immuno-Western blotting using monoclonal antibody to 3-methylcholanthrene-induced cytochrome P-448 (P-448H; m.w.

Derivation of a brain tumor-selective monoclonal antibody from hybridoma between mouse myeloma and rat spleen cells immune to syngeneic glioma.

Fischer 344 (F344) rats hyperimmunized with syngeneic 9L/R3 glioma cells produced antibody selective to glioma cells. Hybridomas prepared from the spleen cells of the immunized rat were cloned, and we obtained a hybridoma clone which produced monoclonal IgM antibody, termed FR77, that showed selectivity to glioma cells. Immunoperoxidase staining of cultured cells revealed that FR77 was reactive with 3 lines of rat glioma cells but not with normal F344 rat fibroblasts.

Monoclonal antibodies against a high spin form of rat cytochrome P-448.

Ten monoclonal antibodies reactive with a high spin form of rat cytochrome P-448 (P-448-H) were obtained from hybridoma clones established by a fusion between P3X63Ag8.653 mouse myeloma cells and spleen cells of a BALB/c mouse hyperimmunized with the cytochrome. One monoclonal antibody recognized an epitope characteristic for P-448-H.

Human bladder cancer cell-surface antigens recognized by murine monoclonal antibodies raised against T24 bladder cancer cells.

Seven hybridoma clones producing monoclonal antibodies (HBJ8, HBJ27, HBJ67, HBJ71, HBJ98, HBJ104 and HBJ127) were selected from hybridomas prepared by fusion between P3 X Ag8.653 mouse myeloma cells and spleen cells of a BALB/c mouse immunized with T24 human urinary bladder cancer cells, and the binding specificity of, and the molecular characters of the antigens defined by, these monoclonal antibodies were examined. The cell-surface antigens detected with these monoclonal antibodies from T24 bladder cancer cells were as follows: 1) HBJ27-defined gp85 antigen common in all human cells or tissues tested, 2) HBJ98- or HBJ127-defined gp125 antigen distributed in all epithelial and non-epithelial human tumor cell lines tested and in basal layers of the skin or esophagus, proximal tubules of the kidney, and crypts of the gastric and intestinal mucosa, 3) HBJ8-defined gp(40/90) and HBJ67-defined gp83 antigens distributed in a characteristic portion of epithelial tumor cell lines, 4) HBJ71-defined antigen of protein nature and HBJ104-defined antigen of unknown character, both being detected from immunizing T24 cells and a few epithelial tumor cell lines.

Monoclonal antibodies against cell surface antigens present on human urinary bladder cancer cells.

After hybridomas were prepared by cell fusion between the mouse myeloma cell line P3x63Ag8.653 and the spleen cells of a BALB/c mouse hyperimmune to the human bladder cancer KU-1 cell line, they were cloned. Monoclonal antibodies (HBA4, HBE3, HBE10, HBF2, HBG9, and HBH8) from the hybridoma clones were selected for their serologic reactivity to cell surface antigens of KU-1 cells and for their unresponsiveness to 2 human lymphoma cell lines.

A proliferation-associated rat cell surface antigen recognized by a murine monoclonal antibody.

B3 murine IgG1 monoclonal antibody against BC47 rat bladder cancer detected an antigen distributed on the cell surface of neoplastic cells and proliferating normal tissue cells. The percentage of B3 antigen-positive cells in lymphocytes was increased by mitogen stimulation. The molecular weight of the antigen was 130,000 or 140,000 daltons.

Monoclonal antibodies to various morphologic components of human skin.

Somatic cell hybrids were established from the mouse myeloma, P3x63Ag8.653 cells, and the spleen cells of a mouse hyperimmune to human epidermal cells. Indirect immunofluorescence test with hybridoma culture fluids displayed that 253 out of 263 hybridoma cultures secreted antibodies reactive with the frozen sections of human skin.

Antitumor effect of actinomycin D entrapped in liposomes bearing subunits of tumor-specific monoclonal immunoglobulin M antibody.

Sonicated liposomes containing actinomycin D in the membranes were chemically coated with the subunits of monoclonal immunoglobulin M (IgM) antibody against a mouse mammary tumor-associated antigen (MM antigen) and examined for their in vitro and in vivo antitumor effects against MM46 (MM+) and MM48 (MM-) tumors of C3H/He mouse origin. The antibody-bearing, actinomycin D-containing liposomes (chemoimmunoliposomes) were selectively bound to MM+ tumor cells and showed much more in vitro cytotoxicity against the tumor cells than that shown by free actinomycin D. The in vivo antitumor effect of the chemoimmunoliposomes was tested on the mammary tumor cells (5 X 10(4) to 5 X 10(6) transplanted i.

Mixed lymphocyte-autologous tumor cell reaction with primary rat bladder tumors: difficulty in the discrimination of tumor-specific antigen.

The presence of tumor-specific or tumor-associated antigens in primary urinary bladder tumors induced by N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in ACI/N rats was examined by means of mixed lymphocyte-tumor cell culture. Bladder tumors (papilloma with cancer foci or cancer) induced by BBN treatment for 18 weeks or more (BT), epithelial cells from papillary hyperplasia induced by 6-week BBN treatment (PH), and normal bladder epithelial cells (NBE) were isolated from the bladder and used as stimulator cells after mitomycin C treatment. BT, PH and NBE all strongly stimulated the blastogenesis of the autologous lymphocytes on culture in medium containing fetal calf serum (FCS), but the spleen cells also stimulated the autologous lymphocytes in this medium.

Emergence of an epidermis-associated cell surface antigen in epithelial cells of the rat urinary bladder in carcinogenesis.

Inbred ACI/rats were treated with a selective urinary bladder carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN). Two cell surface antigens on the epithelial cells of the bladder, normal bladder epithelium (NBE) antigen unique to normal epithelial cells of the bladder and BC-SK antigen specific for bladder cancer and epidermis, were periodically assayed by mixed hemadsorption test with the typing sera against each antigen. Treatment with BBN for 8 or more weeks induced papillomas or cancers that expressed a definite level of BC-SK antigen, whereas a 4-week treatment did not render the bladder cells with a significant level of this antigen.

Cell surface antigens in normal and neoplastic urinary bladder epithelial cells of the rat.

Cell surface antigens of normal urinary bladder epithelium (NBE) and bladder cancers of inbred ACI/N rats were assayed by antisera raised in rabbits. Mixed hemadsorption test combined with absorption analysis defined three kinds of cell surface antigens in both NBE and bladder cancer cells; the first antigen was common to lymphoid cells, the second was present in most of the epithelia, and the third was unique to NBE and present to a lesser extent in bladder cancers. In addition, bladder cancers had a fourth antigen that was common only to epidermis.