Ex vivo perfusion of a tumor-containing colon with monoclonal antibody.
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Adenocarcinoma↗
Biomedical subjects
H Koprowski
Publications and source records attributed to H Koprowski.
Structural characterization of the 'melanoma-specific' antigen detected by monoclonal antibody 691I5Nu-4-B.
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Amino Acids↗
Herpes simplex type 1 DNA in human brain tissue.
Herpes simplex virus type 1 (HSV-1) is known to reside latently in the trigeminal ganglia of man. Reactivation of this virus causes skin lesions and may occasionally infect other tissues, including the brain. To determine whether the brain tissue of humans free of clinical signs of HSV-1 infection contains any trace of HSV-1, we examined the DNA from brain tissue by endonuclease digestion, separation of the fragments by gel electrophoresis, and hybridization with labeled HSV-1 DNA probes. Hybrid bands were detected autoradiographically in experiments using cloned and virion-purified fragments of the HSV-1 genome. HSV-1 DNA sequences were found in 6 of 11 human brain DNA samples tested. In some cases, these bands corresponded to the bands expected for the complete viral genome, whereas others contained bands representing only a part of the genome. In some cases, the terminal fragments could be found, suggesting that the DNA was in a linear, nonintegrated form.
Brain↗
Analysis of a predominant immunoglobulin population in the cerebrospinal fluid of a multiple sclerosis patient by means of an anti-idiotypic hybridoma antibody.
We have produced hybridoma antibodies directed against immunoglobulins present in the cerebrospinal fluid of a patient with multiple sclerosis (MS). One hybridoma antibody recognized an idiotypic determinant of an immunoglobulin population [an idiotype (Id)] which constituted approximately 1% of the immunoglobulin present in the cerebrospinal fluid. The Id focused in the pH range 8.0-8.4. It was present at roughly 10- to 15-fold higher relative (compared to total immunoglobulin) concentration in cerebrospinal fluid than in serum of the homologous MS patient. The Id could not be detected in three cerebrospinal fluid samples and 28 serum samples of heterologous MS patients or in the serum of 43 optic neuritis patients. The Id persisted in the homologous MS patient at increased concentration over the entire (6 years) observation period. The Id could be shown to react with a Theiler murine encephalomyelitis virus strain WW which was isolated from mice after inoculation with periplaque white matter from brain in a histologically confirmed case of MS.
Animals↗
Detection of monoclonal antibody-defined colorectal carcinoma antigen by solid-phase binding inhibition radioimmunoassay.
We have established a solid-phase binding inhibition radioimmunoassay for the detection of colorectal carcinoma-specific antigens in tissue culture supernatants of human colorectal carcinoma cell lines and in serum and urine of colorectal carcinoma patients. Using the [3H]glucosamine-labeled cell membrane glycolipid antigen and colorectal carcinoma-specific monoclonal antibodies in this assay, we have been able to detect several human colorectal carcinoma membrane-specific antigens that are released from the cell membrane into tissue culture supernatants, and an antigen detected by antibodies 1116-NS-19-9 and 1116-NS-52a that is found only in the serum and urine of cancer patients.
Antibodies, Monoclonal↗
Cytotoxic activity and interferon production by lymphocytes from patients with multiple sclerosis.
Peripheral blood lymphocytes from MS patients and from healthy control donors were compared for their ability to mediate spontaneous and antibody-dependent cell-mediated cytotoxicity. They were also compared for their ability to respond to infection with various strains of measles and sSPE viruses with interferon production and enhanced NK activity. Neither SLMC nor ADCC against several different target cells was found to be impaired in the MS population. Furthermore, no defect was detected in the response of patients' lymphocytes to virus challenge in vitro in terms of both activation of NK cells and interferon production. Enhanced NK activity was also induced by an exogenous interferon preparation and by Poly I:C to the same extent in patients and controls.
Adult↗
Release of monoclonal antibody-defined antigens by human colorectal carcinoma and melanoma cells.
Many but not all of the cell surface antigens studied were released into tissue culture medium of human colorectal carcinoma and melanoma cells maintained in vitro. Two groups of monoclonal antibody-defined antigens are described. One group is composed of molecules that are released easily and in large quantitites into the tissue culture milieu. In this group, four glycolipid antigens of colon carcinoma, including a monosialoganglioside, and five glycoproteins of melanoma were detected in tissue culture supernatants. Antigens of the second group could not be detected in tissue culture supernatants. This group included one colorectal carcinoma antigen, the nature of which is unknown, and a glycoprotein of melanoma cells.
Antibodies, Neoplasm↗
Production of human hybridomas secreting antibodies to measles virus.
Monoclonal antibodies against a variety of antigens can be produced using techniques of somatic cell hybridization between cells of rodent myeloma lines and B cells derived from animals immunized against a given antigen. However, because of the monoclonal antibodies secreted by these hybridomas are of rodent origin, their use in human immunotherapy is limited. Thus the production of B-cell hybrids that secrete human monoclonal antibodies may be of considerable value. We have hybridized a hypoxanthine phosphoribosyl transferase (HPRT)-deficient human B-cell line derived from a patient suffering from multiple myeloma with peripheral lymphocytes obtained from a patient with subacute sclerosing panencephalitis (SSPE). These hybridomas were found to secrete human IgM specific for measles virus nucleocapsids.
Antibodies, Viral↗
Hybridomas revisited.
In the report by John C. Behrendt et al. "Aeromagnetic and radio echo ice-sounding measurements show much greater area of the Dufek Intrusion, Antarctica" (29 Aug., p. 1014), the word "expedition" should have read "exploitation" in line 13 of the first paragraph on page 1014. Also, in line 2 of the next to last paragraph on page 1016, "50 to 60 cm/sec(2)" should have read "50 to 60 (cm sec(2)) x 10(-3)."
Animals↗
Antigenic variants of rabies virus.
Antigenic variants of CVS-11 strain of rabies virus were selected after treatment of virus populations with monoclonal antibodies directed against the glycoprotein antigen of the virus. These variants resisted neutralization by the hybridoma antibody used for their selection. Two independently mutating antigenic sites could be distinguished when five variants were tested with nine hybridoma antibodies. The frequency of single epitope variants in a cloned rabies virus seed was approximately 1:10,000. Animals were not or only partially protected when challenged with the parent virus or with another variant, but were fully protected against challenge with the virus used for immunization. Variants were also detected among seven street viruses obtained from fatal cases of human rabies. Animals immunized with standard rabies vaccine were protected against challenge with some but not all street rabies variants. A comparative antigenic analysis between vaccine strain and challenge virus by means of monoclonal antiglycoprotein antibodies showed a slightly closer degree of antigenic relatedness between vaccine and challenge strain in the combinations where vaccination resulted in protection. It remains unknown, however, whether these apparently minor antigenic differences in the glycoproteins account for the varying degrees of protection. The results of this study clearly indicate that the selection of vaccine strains and the methods used to evaluate the potency of rabies vaccines need to be revised.
Animals↗
Production of monoclonal antibodies in serum free medium.
We have succeeded in growing several established hybridoma cell lines (1116NS-19; 116NS-33a; 1116NS-52a; 1083-17-1A; 691-19-19) in a serum free medium supplemented with physiological concentrations of insulin (5 microgram/ml) and transferrin (5 microgram/ml). The hybridoma cells replicate in this medium and will secrete monoclonal antibodies in quantities comparable to those produced in the presence of serum. Adapted hybridoma cultures have been secreting antibodies in the supplemented serum free (SSF) medium for more than 3 months. Parental mouse myeloma P3x63Ag8 cells and non-secreting subclone 653 cells cannot survive longer than 6 days under the same conditions. Both insulin and transferrin are necessary for hybridoma cells to grow in serum free medium and omission of either one will not permit cell multiplication.
Antibody-Producing Cells↗
Biochemical characterization of human melanoma cell surfaces: dissection with monoclonal antibodies.
This report describes the structures of three distinct human melanoma surface antigens detected by monoclonal anti-melanoma antibodies. One antigen, expressed on all melanoma cells and on certain astrocytoma cells but not on any other kind of normal or tumor cell tested, has been shown to consist of our associated polypeptide chains. A second antigen expressed on many but not all melanomas has been identified as the antigenic product of the HLA-D locus, the DR antigen. A third protein antigen is not found on any normal cell tested but occurs on some, but not all, tumors of various origins.
Antibodies, Neoplasm↗
Antibody-directed cytotoxic agents: use of monoclonal antibody to direct the action of toxin A chains to colorectal carcinoma cells.
We have constructed cell-specific cytotoxic agens by covalently coupling the A chain from diphtheria toxin or ricin toxin to monoclonal antibody directed against a colorectal carcinoma tumor-associated antigen. Antibody 1083-17-1A was modified by attachment of 3-(2-pyridyldithio)propionyl or cystaminyl groups and then treated with reduced A chain to give disulfide-linked conjugates that retained the original binding specificity of the antibody moiety. the conjugates showed cytotoxic activity for colorectal carcinoma cells in culture, but were not toxic in the same concentration range for a variety of cell lines that lacked the antigen. Under defined conditions virtually 100% of antigen-bearing cultured cells were killed, whereas cells that lacked the antigen were not affected. Conjugates containing toxin A chains coupled to monoclonal antibodies may be useful in studying functions of various cell surface components and, possibly, as tumor-specific therapeutic agents.
Animals↗
Use of hybridoma monoclonal antibodies in the detection of antigenic differences between rabies and rabies-related virus proteins. II. The glycoprotein.
Twenty-five hybridoma cultures secreted monoclonal antibodies directed against the glycoprotein of rabies or rabies-related viruses. The antibodies had different specificities for the glycoproteins of eight rabies and rabies-related viruses. They could be classified into fourteen groups which probably correspond to different antigenic determinants on the glycoproteins. These hybridomas when used in either radioimmunoassay (RIA) or in neutralization tests allow differentiation of laboratory strains of rabies virus from each other as well as from the rabies-related viruses.
Animals↗
Use of hybridoma monoclonal antibodies in the detection of antigenic differences between rabies and rabies-related virus proteins. I. The nucleocapsid protein.
Twenty-one hybridoma cultures, obtained through the fusion of mouse myeloma cells with splenocytes of BALB/c mice immunized with either rabies virus or Mokola virus, secreted monoclonal antibodies specific for the nucleocapsid of the inducer virus. They displayed different specificities for the nucleocapsids of rabies and rabies-related viruses and could be classified into eight groups which are likely to correspond to different antigenic determinants on the nucleocapsid. Four strains of fixed rabies virus (CVS, ERA, Flury-LEP and Kelev) could not be differentiated by the nucleocapsid-specific hybridoma antibody. The Flury-HEP virus (derived from Flury-LEP) as well as the rabies-related viruses Mokola, Lagos bat and Duvenhage, showed marked differences in their reactivities with hybridoma antibodies to nucleocapsid. A selected panel of three of these hybridomas may be used for a rapid differential diagnosis among all members of the Lyssavirus group.
Animals↗
Variations in herpes simplex virus isolated from human ganglia and a study of clonal variation in HSV-1.
Growth of human sensory ganglia in culture has led to the reactivation of herpes simplex virus from over 50% of cases studied. Infected cell polypeptide and restriction enzyme analysis has led to the conclusion that each individual has one unique latent strain of HSV-1 that can be present in more than one ganglion in the body. Analysis of 91 isolates has shown that the long region of the genome is variable in terms of DNA restriction sites, DNA sequences, and in coding for the majority of variable polypeptides. The short region is stable with only polypeptides Vmw 21 and 22, restriction sites HindIII-(M-N) and BglII-(G-H) and the DNA sequence BamHI-1' having been found to vary. The insertion and deletion of small DNA sequences at specific locations allows individual reactivation events to be distinguished. Detection of information able to complement and produce ts+ virus on superinfection of otherwise negative ganglia with ts mutants, has led to the conclusion that ganglion cells may harbor herpes virus-related information that is only detectable by use of such genetic probes.
Base Sequence↗
Identification of a unique idiotype in cerebrospinal fluid and serum of a patient with multiple sclerosis.
Anti-idiotypic antibodies were prepared in mouse ascites fluid against the CSF-IgG of a patient with multiple sclerosis. After adsorption with pooled human IgG, the ascites fluid antibodies precipitated 20% of labeled autologous CSF IgG. By using a competitive radioimmunoassay, less than one microgram of unlabeled CSF IgG produced 50% inhibition of binding autologous 125I-labeled CSF IgG, whereas 50 micrograms of normal HIgG was not inhibitory. The idiotype could be found in both serum and CSF IgG and persisted over a 5-year period. The absolute concentration of idiotype in the CSF varied somewhat but remained from 4 to 10 times greater than that of the serum. One of 14 heterologous MS CSF was found to contain small amounts of inhibitory protein; eight CSF from patients with other neurologic diseases did not contain the idiotype.
Animals↗