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Biomedical subjects

H Koprowski

Publications and source records attributed to H Koprowski.

At least 289 records · Page 16Linked to original sources
PubMed1984

Phenotypic and functional characterization of T cell clones derived from the cerebrospinal fluid of multiple sclerosis patients.

We describe here T cell cultures and clones established from the cerebrospinal fluid (CSF) of three patients with multiple sclerosis (MS) and one chronic meningitis patient with pleocytosis. Most of the cultures were activated with phytohemagglutinin (PHA) before growth in mitogen-free interleukin 2 (IL 2), and were never restimulated. Some of the clones obtained have been propagated for over 1 yr and are strictly IL 2-dependent. Immunofluorescence analysis performed with various monoclonal antibodies revealed that the CSF-derived lines had the characteristics of activated T cells with a stable expression of either suppressor/cytotoxic or helper/inducer surface antigens. Most of the clones established had a predominantly suppressor phenotype (OKT8+), except for the clones derived from one MS patient, which expressed only the helper phenotype (anti-Leu-3a+). Consistent with these data, the CSF-derived cultures displayed a variety of immunoregulatory functions, such as the ability to lyse nonspecific and PHA-stimulated target cells, to produce IL 2 upon mitogenic activation, and to modulate polyclonally induced Ig responses. The availability of long-term CSF T cell cultures derived from MS patients at various disease stages might provide a useful tool in investigating the factor(s) involved in the etio-pathogenesis of the disease.

Adolescent↗
PubMed1984

Identification of melanoma-associated antigens using fixed tissue screening of antibodies.

Early culture supernatants from hybridomas that were obtained through fusions of mouse myeloma cells with lymphocytes of melanoma-immunized mice were screened for their reactivity with a paraffin-embedded cell block of a melanoma cell line, using a biotin:avidin immunoperoxidase procedure. Eleven monoclonal antibodies were derived that define several new melanoma-associated antigens. The antigens include a neutral glycolipid, gangliosides, membrane-associated proteins, cytosolic proteins, and strongly secreted proteins. These antibodies, which detect antigens that withstand tissue fixation and embedding procedures, were tested for reactivity in fixed cell lines, as well as in melanoma biopsies. These antibodies may provide powerful tools in diagnostic studies of human malignant melanoma biopsy material.

Animals↗
PubMed1983

A study of gastrointestinal cancer-associated antigen (GICA) in human fetal organs.

The gastrointestinal cancer-associated antigen (GICA) characterized by 1116 NS 19-9 monoclonal antibody was studied in human fetal organs by immunohistology and immunofixation on nitrocellulose sheets. It was found in all the gastrointestinal tracts of fetuses and newborns. Other fetal organs, except biliary and pancreatic ducts, were negative. Immunohistological data and enzymatic studies led us to conclude that GICA is predominantly a mucin-type glycoprotein in fetal organs.

Antibodies, Monoclonal↗
PubMed1983

Human macrophages armed with murine immunoglobulin G2a antibodies to tumors destroy human cancer cells.

Macrophages isolated from tumor-bearing patients as well as cultured human monocytes express Fc receptors that cross-react strongly with murine immunoglobulins of the G2a but only slightly or not at all with the G1, G2b, or G3 subclasses. Such macrophages in the presence of murine immunoglobulin G2a monoclonal antibodies to tumors mediated the killing of tumor cells in vitro. These data suggest that monoclonal antibodies of the G2a subclass may be useful in the immunotherapy of human cancer.

Animals↗
PubMed1983

A simple procedure for determining Lewis phenotypes in human saliva.

We have selected 4 murine monoclonal antibodies that specifically bind to Lewis type blood group haptens to establish an assay in which antigens present in human saliva are bound to polystyrene beads and then detected by monoclonal antibodies in radioimmunoassay or in peroxidase-antiperoxidase immunoassay. Two of these antibodies detect both the Lea and Leb determinants (Leab), 1 antibody detects only Lea, and 1 only Leb. All Lewis phenotypes (Lea-b-, Lea+b-, Lea-b+ and Lea+b+) are easily detectable in this assay. Of the 60 individuals tested, 1 was Lea+b+, 4 Lea-b-, 12 Lea+b- and 43 Lea-b+.

Animals↗
PubMed1983

Mouse monoclonal antibodies against human cancer cell lines with specificities for blood group and related antigens. Characterization by antibody binding to glycosphingolipids in a chromatogram binding assay.

Solid phase radioimmunoassay and a chromatogram binding assay were used to characterize the binding specificities of five monoclonal antibodies generated from mice immunized with human tumor cell lines when tested against various glycolipids. Four antibodies derived from mice immunized with pancreatic carcinoma cells detected specifically the human blood group B determinant, Gal alpha 1 leads to 3Gal (2 comes from 1 alpha Fuc). These antibodies preferred type 2 (Gal beta 1 leads to 4GlcNAc) glycolipids. No reactivity was detected with a rat B determinant based on GalNAc. An antibody derived following immunization with a rectal carcinoma cell line was shown to have binding properties identical with those of an antibody that reacts specifically with the stage-specific embryonic mouse antigen (SSEA-1), bearing the determinant Gal beta 1 leads to 4GlcNAc (3 comes from 1 alpha Fuc) (Gooi, H. C., Feizi, T., Kapadia, A., Knowles, B. B., Solter, D., and Evans, M. J. (1981) Nature 292, 156-158). Thin layer chromatography was used to detect the binding of a monoclonal anti-tumor antibody recently shown to react with a sialylated Lea glycolipid (Magnani, J. L., Nilsson, B., Brockhaus, M., Zopf, D., Steplewski, Z., Koprowski, H., and Ginsburg, V. (1982) Fed. Proc 41, 898) and an anti-Leb antibody (Brockhaus, M., Magnani, J. L., Blaszczyk, M., Steplewski, Z., Koprowski, H., Karlsson, K.-A., Larson, G., and Ginsburg, V. (1981) J. Biol. Chem. 256, 13223-13225) to mixtures of glycolipids from normal and tumor tissues.

Adenocarcinoma↗
PubMed1983

Monoclonal antibodies derived from immunosuppressed mice grafted with human melanoma.

Hybridoma cells producing monoclonal antibodies against tumor-shed antigens were generated by fusing mouse myeloma cells with spleen cells from immunosuppressed mice bearing human melanoma xenografts. Thirty-eight fusion experiments were performed at different stages of tumor growth. Hybridomas producing anti-melanoma antibodies were obtained from 12 spleens in mice bearing 1-4-week-old tumors but at later stages of tumor growth, no hybridomas whatsoever could be obtained. However, the sera of all mice tested showed anti-melanoma antibody binding reactivity at the time of fusion. Using radioimmunoassay (RIA) to select specific antibody secreting hybridoma cultures, the majority of cultures were found to produce antibodies which bound to both 3 M KCl melanoma extracts and melanoma culture supernatants. No stable cultures secreting membrane-reactive (live tumor cell targets) antibodies could be obtained. All of the monoclonal antibodies bound not only to melanoma target cell preparations, but also to preparations from tumors of other origins and only 3 did not bind to normal human fibroblasts. The crossreactivity pattern of binding was confirmed in immunoperoxidase (IP) assays by binding to human tissue sections. The immunosuppressed mouse bearing human tumor xenografts has proven a useful system for production of monoclonal antibodies against antigens shed by tumor cells.

Animals↗
PubMed1983

Mass spectrometry of a human tumor glycolipid antigen being defined by mouse monoclonal antibody NS-19-9.

With an antibody-to-chromatogram binding assay to follow the preparation a glycolipid was isolated from human pancreatic carcinoma using a mouse monoclonal antibody of apparent specificity for gastrointestinal tumors. Direct probe mass spectrometry of three derivatives established the sugar sequence as NeuAc-hexose-(fucose)N-acetylhexosamine-hexose-hexose and the ceramide to be composed mainly of phytosphingosine and 16-24 carbon 2-hydroxy fatty acids. NMR spectroscopy of two of the derivatives made likely the presence of the sequence Gal beta 1 leads to 3GlcNAc(4 comes from 1 alpha Fuc)beta 1 leads to, which is the blood group Lewis a determinant. This is in agreement with recent results from degradation studies.

Adenocarcinoma↗
PubMed1983

Embryonic precancerous and cancerous human antigens recognized by monoclonal antibodies.

Monoclonal antibodies produced after immunization of mice with human melanomas define protein antigens expressed not only by melanomas by also by other tumours of neural crest origin such as astrocytomas and neuroblastomas. Other monoclonal antibodies react with antigens expressed by melanomas and fetal but not adult human melanocytes. Cells of common naevi and of precancerous lesions such as dysplastic naevi share many antigens with melanomas but not with normal melanocytes. Unlike melanomas, naevi in tissue culture are characterized by a finite lifetime. Factors that are instrumental in malignant transformation of dysplastic naevi in vivo and are apparently lacking in the tissue culture system are currently under study. Monoclonal antibodies produced after immunizing mice with cells of human gastrointestinal carcinomas define glycolipid antigens. The carbohydrate structure of one of these antigens is lacto-N-fucopentaose III. This antigen is very strongly expressed by a variety of human tumours and in the immunoperoxidase assay the respective monoclonal antibody also binds to normal human epithelium. Another monosialoganglioside is an antigen expressed by cells of gastrointestinal tract tumours such as of the pancreas, stomach and large bowel and by cells of villous adenomas of the colon, but not by cells of normal colonic mucosa. Its carbohydrate is a sialylated Lea-active pentasaccharide (sialylated lacto-N-fucopentaose II). A hitherto undiscovered sialyltransferase, which may be involved in synthesis of this antigen from Lewis A glycolipid, is probably active in tumour tissue and at early stages of embryogenesis but not in normal adult tissue. Human tumours implanted in mice are destroyed by monoclonal antibodies showing binding specificities for the implanted tumour. Only monoclonal antibodies of IgG2a isotype show tumoricidal activity. Destruction of the tumour is mediated by macrophages which adsorb the IgG2a monoclonal antibody to an Fe receptor. The tumours can also be destroyed, in the presence of monoclonal antibodies, by human monocytes, which after maintenance in culture fore two weeks develop Fc receptors for mouse IgG2a antibody.

Animals↗
PubMed1983

Isolation and chemical characterization of a melanoma-associated proteoglycan antigen.

Many melanoma-associated antigens have been identified by monoclonal antibodies. One of these monoclonal antibodies, O1-94-45, binds only to melanomas, nevus cells, some astrocytomas, and fetal epitheloid cells. There are approximately 100,000 cell surface antigens per melanoma cell with an association constant of 3 X 10(8) M-1. The antigen is efficiently extracted from the membrane only in the presence of detergent and is, therefore, bound by hydrophobic forces. However, it is also shed into the culture supernatant during normal cell growth. The two components of the O1-95-45 antigen are a chondroitin sulfate proteoglycan (CSP, greater than 500,000 Da) and a glycoprotein gp260 (260,000 Da, pI 6.9). CSP contains chondroitin sulfate and N-linked and O-linked oligosaccharides. Only N-linked saccharides were associated with gp260. The antigenic site is expressed on both components and is heat-sensitive. Since the CSP was converted to gp260 by chondroitinase, the protein cores of the two molecules are the same or similar. For more detailed study the O1-95-45 antigen was purified by immunoaffinity chromatography. The amino acid composition of the purified antigen was relatively polar with an unusually high Leu content and low Lys content. Initial attempts to sequence the antigen were unsuccessful probably due to a blocked N-terminus. CSP and gp260 were partially separated by gel filtration chromatography, and both were found to carry the O1-95-45 antigenic determinant. Three other monoclonal antibodies were found to bind the purified antigen at a site or sites different from the O1-95-45 epitope and one other monoclonal antibody may bind at the same site. Two of these antibodies were used for a double determinant immunoassay.

Amino Acids↗
PubMed1983

Identification and isolation of a common tumor-associated molecule using monoclonal antibody.

A monoclonal antibody, 16B-13, derived from the immunization of BALB/c mice with a lung tumor line, immunoprecipitates a common tumor-associated molecule with an apparent mol. wt of 37,000 from lactoperoxidase-iodinated lung carcinoma, colon carcinoma, gastric carcinoma, brest carcinoma, melanoma and lymphoma cells, but not from normal fibroblasts. Analysis by two-dimensional gel electrophoresis of the cell surface-labeled 16B-13 antigen from a colorectal and a melanoma cell line reveals four components with similar mol. wts but with different isoelectric points. The antigen purified from a colorectal carcinoma cell line by immunoaffinity chromatography was shown to be a 37,000 mol. wt polypeptide similar to that obtained by the lactoperoxidase-labeling procedure. However, the purified antigen from the melanoma cell line shows the presence of a 65,000 mol. wt polypeptide and the loss of the 37,000 mol. wt component as detected by Coomassie blue staining and immunoprecipitation.

Animals↗
PubMed1983

Characterization of an antigenic determinant of the glycoprotein that correlates with pathogenicity of rabies virus.

The pathogenicity of fixed rabies virus strains for adult mice depends on the presence of an antigenic determinant on the viral glycoprotein. Two virus-neutralizing monoclonal antibodies have been used to identify this determinant. All pathogenic strains of fixed rabies virus bind to these antibodies and are neutralized by them, whereas nonpathogenic strains fail to react with these monoclonal antibodies and are not neutralized by them. Antigenic variants of the rabies virus with altered glycoprotein were selected by growing virus in the presence of one monoclonal antibody, 194-2. All variants that lost their ability to react with this antibody and an additional antibody, 248-8, were found to be nonpathogenic for adult mice. Analysis of tryptic peptides of the glycoproteins of pathogenic parent virus and nonpathogenic variants and the amino acid sequence of a specific variant tryptic peptide revealed that the change in pathogenicity corresponded to an amino acid substitution at position 333 of the glycoprotein molecule. The nucleotide sequence of the nonpathogenic variant glycoprotein gene contained a base change that confirmed the single amino acid substitution in the tryptic peptide replacing arginine-333 in the parental glycoprotein. We conclude that arginine-333 is essential for the integrity of an antigenic determinant and for the ability of rabies viruses to produce lethal infection in adult mice.

Amino Acid Sequence↗
PubMed1983

Molecular mimicry in virus infection: crossreaction of measles virus phosphoprotein or of herpes simplex virus protein with human intermediate filaments.

Using monoclonal antibodies, we demonstrate that the phosphoprotein of measles virus and a protein of herpes simplex virus type 1 crossreact with an intermediate filament protein of human cells. This intermediate filament protein, probably vimentin, has a molecular weight of 52,000, whereas the molecular weights of the measles viral phosphoprotein and the herpes virus protein are 70,000 and 146,000, respectively. Crossreactivity was shown by immunofluorescent staining of infected and uninfected cells and by immunoblotting. The monoclonal antibody against measles virus phosphoprotein did not react with herpes simplex virus protein and vice versa, indicating that these monoclonal antibodies recognize different antigenic determinants on the intermediate filament molecule. The significance of these results in explaining the appearance of autoantibodies during virus infections in humans is discussed.

Antibodies, Monoclonal↗
PubMed1983

Detection of carcinoembryonic antigen and related antigens in sera of patients with gastrointestinal tumors using monoclonal antibodies in double-determinant radioimmunoassays.

Of 14 monoclonal antibodies produced in six different laboratories, 13 bound to purified preparations of carcinoembryonic antigen (CEA). All antibodies reacted to spent medium of colorectal carcinoma cell lines. Competitive binding studies indicated that 12 different antigenic determinants representing six different groups were detected on the CEA molecule(s). Six antibodies were used in double determinant radioimmunoassays (RIA) to detect CEA and CEA-related antigens in sera of 311 patients with various gastrointestinal diseases and of normal donors. None of up to 115 sera of healthy donors had elevated antigen levels with four out of the six monoclonal antibodies tested, whereas up to 9% of sera showed elevated antigen levels when tested with two antibodies. Between 1.4% and 4.4% of sera from patients with inflammatory and benign neoplastic diseases of the gastrointestinal tract were positive. Antigen levels were elevated in 56 to 75% (depending on antibody used) of sera from patients with advanced gastrointestinal tumors. These preliminary results indicate that double-determinant immunoassays with a panel of monoclonal antibodies might improve conventional CEA assays by reducing the number of false positive sera detected by polyclonal sera in patients with benign inflammatory bowel diseases.

Antibodies, Monoclonal↗
PubMed1983

CSF antibodies to myelin basic protein and oligodendrocytes in multiple sclerosis and other neurological diseases.

Cerebrospinal fluid (CSF) from 18 multiple sclerosis (MS) patients, 13 subacute sclerosing panencephalitis (SSPE) patients, 22 other neurological disease (OND) patients, and 7 neurotic patients as controls were tested in an 125I-labeled anti-human F(ab')2 binding assay for the presence of antibodies to normal human brain cells from tissue culture, human fibroblasts, plasma membranes of MS and normal human brain, myelin basic protein (MBP) and bovine oligodendrocytes. Antibodies to MBP and to oligodendrocytes were found in the CSF of MS, SSPE and OND patients. Absorption of CSF with bovine CNS myelin significantly diminished binding activity to oligodendrocytes. Antibodies in the CSF against MBP and oligodendrocytes, on which some myelin determinants are expressed, seem to be a common feature of diseases in which demyelination is a component.

Antibodies↗
PubMed1983

Search for Sendai 6/94 viral RNA in the antigen-free cell line Cl-C-2 isolated from human multiple sclerosis brain tissue.

The viral antigen-free cell line Cl-C-2, obtained from multiple sclerosis brain tissue by cell fusion with CV-1 cells, was examined for the presence of intracellular virus-specific RNA sequences of the persistent Sendai 6/94 virus by nucleic acid hybridization. As a specific probe for this assay, an in vitro synthesized cDNA was used. Oligodeoxyguanidylic acid served as a primer for the initiation of cDNA synthesis. The 6/94 RNA was detectable as expected in the viral antigen-expressing cell lines Cl-E-8 and Cl-F-2, which were used as a reference of the same source. In the viral antigen-free cell line Cl-C-2, however, no viral RNA sequences have been found by hybridization experiments. Corresponding superinfection studies confirmed the conclusion that in cell line Cl-C-2 no viral components are present. The lack of expression of viral proteins and of protection against superinfection seems to be correlated with the lack of viral RNA in Cl-C-2 cells, which may eliminate the persistent virus by a cellular defense mechanism.

Animals↗