Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0026764 (multiple myeloma)
 36,148 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Hybridoma cell lines secreting monoclonal antibodies (Mab) directed against the products formed by reaction of alkylating N-nitroso carcinogens with DNA have been established by fusion of rat or mouse myeloma cells, respectively, with spleen cells of rats or mice immunized either with conjugates of various alkyl-ribonucleosides with suitable carrier proteins, or with alkylated DNA electrostatically complexed to carrier proteins. Due to their high affinity and specificity, some of these Mab detect very low amounts of the respective alkyl-deoxynucleosides (e.g., O6-methyl-2'-deoxyguanosine, O6-ethyl-2'-deoxyguanosine, O6-n-butyl-2'-deoxyguanosine, O6-isopropyl-2'-deoxyguanosine, O4-methyl-2'deoxythymidine, O4-ethyl-2'-deoxythymidine) and can be used in various types of immunoassays. With a competitive radioimmunoassay (RIA), specific DNA alkylation products can be quantitated in hydrolysates of cellular DNA, in body fluids, or in urine. The RIA is routinely applicable, reproducible, and sufficiently sensitive to permit the quantitation of femtomole amounts of modified nucleosides in small samples of DNA. When the alkyl-deoxynucleosides in question are separated from bulk DNA by high-performance liquid chromatography prior to analysis by RIA, very low levels of modification in DNA can be detected. The immuno-slot-blot (ISB), a noncompetitive solid-phase immunoassay, is more sensitive than the RIA. For analysis by ISB, alkylated DNA is heat-denatured and immobilized on nitrocellulose filters prior to exposure to the respective Mab and subsequent binding of a second (125I-labelled or biotinylated) antibody. In immunocytological analysis (ICA), the binding of Mab to alkyl-deoxynucleosides is visualized in individual cells by immunostaining of denatured nuclear DNA in situ (direct immunofluorescence; peroxidase-staining).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Monoclonal antibody-based immunoanalytical methods for detection of carcinogen-modified DNA components. 353 92

Legionellae are widely spread in natural and man-made habitats. In many instances contaminated tap water has been linked to sporadic or endemic cases of human pulmonary infections, but it is not known why, in spite of frequent occurrence, legionellae only rarely cause disease. Monoclonal antibodies against Legionella pneumophila serogroup 1 (Philadelphia 1) were prepared in order to distinguish between subtypes of this serogroup. Balb/c mice were immunized i.v. three times with heat inactivated bacteria. Antibody formation was detected by an enzyme-linked immunosorbent assay (ELISA) technique using peroxidase-conjugated antimouse IgG. Spleen cells were then fused with NS-1 myeloma cells and cloned by limiting dilution. Four monoclonal antibodies were studied in detail. The study included 47 strains of L. pneumophila: 19 strains were of human origin and 28 were isolated from different environmental sources. Most were from tap water, but none from natural habitats. All strains belonged to serogroup 1 as defined by direct immunofluorescence (DFA) using monospecific FITC-labelled polyclonal antisera from rabbits. The strains were further characterized by beta-lactamase production, activity of catalase, oxidase and proteases, analysis of ubiquinones, and demonstration of membrane protein patterns by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. A strong homogenicity between all the strains could be revealed by these methods independent of their origin. One of the monoclonal antibodies (B-1) was able to distinguish between human and environmental isolates. Eighteen of the 19 human strains reacted very strongly in DFA using antimouse immunoglobulin. No reaction, however, was seen with all of the environmental strains. Immunoblots were performed for characterization of the distinguishing feature using membrane complexes of all strains on nitrocellulose strips. The blots were incubated with antibody B-1, and immune complexes were detected by 125I-protein A. Broad intense blackening was seen between 22 and 70 kilodalton. This result suggests that no single protein, but rather a smaller component such as an oligosaccharide attached to constituents of different molecular weights, might be responsible for the discriminating reaction.
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PMID:Discrimination between clinical and environmental strains of Legionella pneumophila by a monoclonal antibody. 353 65

The protein B of group B streptococci can bind in a nonimmune reaction to Ig of the IgG and IgM classes of various mammalian species (i.e., human, mouse, rabbit, and bovine). Protein B binding involves the Fc parts of both IgG and IgM molecules. Monoclonal or polyclonal IgG or IgM and the IgM-FC5 mu fragment of human myeloma protein combined with the protein B thereby inhibiting protein B-induced hemolysis in the CAMP reaction. The protein B/Ig complex can be dissociated with 1% Triton or guanidine-HCl (6 M). Mice infected intraperitoneally with sublethal doses of group B streptococci (GBS) and that received seven repeated intravenous injections of highly purified protein B during the first 9 h of infection developed fatal septicemia within 24 h with colony counts of up to 10(8) CFU/ml in the blood. Animals treated in the same way with either PBS or trypsinized protein B recovered. The protein B itself was not pathogenic when injected into healthy mice. Tissue sections of liver or spleen from mice infected with a lethal dose of GBS revealed the presence of protein B together with large numbers of cocci when stained by the peroxidase method using specific antibodies raised against purified protein B in the rabbit.
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PMID:Unspecific binding of group B streptococcal cocytolysin (CAMP factor) to immunoglobulins and its possible role in pathogenicity. 354 80

About 700 antibody-secreting hybrids were obtained by fusion of lymphocytes, (harvested from mice hyperimmunized with the human gastric carcinoma line KATO III) and P3-X63-Ag8-653 myeloma cells. Antibody specificity was screened in ELISA performed on glutaraldehyde-fixed cultured cells and on paraffin-embedded tissue sections stained with the method of avidin-biotin-peroxidase. When tested in ELISA, the monoclonal antibody produced by the hybrid clone BD-5 was found to bind only to the cell line used as immunogen, among the many neoplastic or normal human cell lines tested. When assayed on paraffin sections with the avidin-biotin-peroxidase method, the BD-5 monoclonal antibody stained gastric carcinomas, but not the normal mucosa. Pancreatic carcinomas were also stained, while the corresponding normal gland was not. The antibody strongly stained the normal colonic and small intestinal mucosa. Among the other normal or neoplastic tissues tested, a weak reactivity was observed only with some epithelial cells of the salivary glands and with some carcinoma cells of the uterus and of the lung. It is concluded that the BD-5 antibody reacts with an epitope normally present on intestinal mucosa, which, following neoplastic transformation, is ectopically expressed also on gastric and pancreatic carcinomas. This monoclonal could represent a useful reagent for histopathological diagnosis.
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PMID:Production of monoclonal antibodies for the immunohistochemical detection of gastric carcinomas. 355 23

A procedure is described for the production of calcitonin-specific hybridomas which involves primary and secondary immunization of Balb/c mice in the hind footpads with free, synthetic human calcitonin and cell fusion of lymphocytes from popliteal lymphonodes with a P3 x 63 myeloma line. This protocol offers the following advantages: (a) it is short and easy to perform, (b) it requires small amounts of unconjugated antigen, and (c) it gives a high yield of antigen-specific IgG-secreting hybridomas. Routine screening was carried out by ELISA on solid phase calcitonin and binding of the monoclonals to free antigen was studied with calcitonin linked to biotin through a 13 carbon atom spacer. Monoclonal couples capable of simultaneously binding calcitonin in solution were found by pairing in all possible combinations 25 purified antibodies, in their unlabelled and biotinylated form, in a checkerboard matrix experimental system. Of the over 30 positive pairs identified, four were used in a one-step enzyme immunoassay for calcitonin determination on microtiter plates in a concn range between 0.1 and 5 ng/ml. With the detecting monoclonal directly conjugated to peroxidase with a heterobifunctional crosslinker, the range of the assay with one monoclonal pair was between 25 and 1000 pg/ml with an 18 hr incubation at 4 degrees C.
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PMID:Production of monoclonal antibodies to calcitonin and development of a two-site enzyme immunoassay. 369 66

To analyze the humoral immune response to melanoma, human-mouse hybridomas were generated by the fusions of regional lymph node lymphocytes of patients with the mouse myeloma cell line M5. Six stable hybridomas were cloned from six separate lymphocyte parents obtained from three patients. Ascites were obtained from nude mice after i.p. injection with cultured hybridoma cells. The monoclonal antibodies, four immunoglobulin Gs and two pentameric immunoglobulin Ms, were partially purified to remove mouse immunoglobulin and then conjugated to biotin for immunocytochemical and immunohistochemical studies. With the avidin:biotin:peroxidase complex method to detect and amplify binding by the biotin-conjugated human monoclonal antibodies, we found the six antibodies to be reactive against cytoplasmic determinants in five short-term melanoma cultures and formalin-fixed paraffin-embedded melanoma tumors from four patients. The antigenic target of the antibodies identified was not carcinoembryonic antigen. Two antibodies, 2-139-1 and 6-26-3, were studied in more detail. Each stained 25 of 25 specimens of melanomas. Little or no reactivity was detected against fixed sections of normal skin, which included tissues such as epidermis, dermis, monocytes, lymphocytes, and vascular endothelium. More striking was the absence of binding to melanocytes in the basal layer of the skin or to pigmented nevus cells. Both antibodies showed cross-reactivity against other tumors, in particular colonic and prostatic carcinomas. In the normal colon, reactivity was restricted to the surface of the columnar epithelium; no reactivity was detected against normal prostatic epithelium. Reactivity was also not observed against liver and lung. However, the epithelia of the renal tubules, pancreatic ducts, and salivary ducts were all reactive. These human monoclonal antibodies identify cytoplasmic melanoma-associated tumor antigens that appear different from the membrane antigens defined by serological approaches and by most mouse monoclonal antibodies.
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PMID:Human monoclonal antibodies directed against melanoma tumor-associated antigens. 369 90

From a surface active fraction of porcine lung lavage fluid, separated by discontinuous sucrose density gradient ultracentrifugation, a protein with a nominal molecular weight (MW) of 15,000 daltons was isolated by sequential extraction with several buffers, including one containing deoxycholate. A monoclonal antibody was prepared from a hybrid cell (8B5E) obtained by fusing a myeloma cell, X63.Ag8.653, with spleen cells of BALB/c mice immunized with the protein. With immunoblotting technique, the antibody was found to be specific to the 15,000 dalton protein and did not react with another surfactant-associated protein with a nominal MW of 38,000 daltons. The antibody's IgG subclass was IgG1 and the light chain was kappa. In immunohistochemical studies using biotinylated antibody, peroxidase reaction products were localized selectively at inclusions of alveolar wall cells which were located chiefly at the alveolar corners. These results strongly suggest that this 15,000 dalton protein was localized in inclusions of alveolar wall cells and did not originate from other larger surfactant-associated proteins degraded after secretion into alveolar space.
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PMID:A monoclonal antibody to the 15,000 dalton protein associated with porcine pulmonary surfactant. 375 1

We have assessed the tumoricidal potential of enzyme-antibody conjugates on murine myeloma cells. Conjugates of glucose oxidase (EC 1.1.3.4) and lactoperoxidase (EC 1.11.1.7) were specifically targeted on the NSO tumor cells. Optimal conditions for tumor cell killing, as assayed by [51Cr] release required the binding of both antibody conjugates to the cell membrane. This is followed by washing and incubation in medium containing glucose and 0.1 mM iodide. Under these conditions 90% of the incorporated [51Cr] labeled is released from the cells, and NSO clonogenicity is reduced by a factor greater than 5 logs by 2 h of incubation.
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PMID:In-vitro cytolysis of myeloma tumor cells with glucose oxidase and lactoperoxidase antibody conjugates. 376 10

In an effort to develop new approaches to the study and control of infectious diarrhea, we prepared murine monoclonal antibodies to the Escherichia coli heat-stable enterotoxin (STa). The toxin was purified from E. coli culture media and conjugated to bovine serum albumin. The STa-bovine serum albumin conjugate was used to immunize BALB/c mice, and the immune spleen cells from these mice were fused with SP2/0 myeloma cells. Resultant hybridomas were screened in an enzyme-linked immunosorbent assay protocol against 500 ng of STa-bovine serum albumin bound to microtiter wells as the solid-phase antigen. Five stable clones were selected and grown further in ascites fluid, which demonstrated anti-STa activity at dilutions of up to 1:500,000 in the enzyme-linked immunosorbent assay for heat-stable enterotoxin. In a competitive enzyme-linked immunosorbent assay format, the antibodies recognized several human and porcine strains of STa to various extents, but did not recognize E. coli heat-labile toxin, cholera toxin, or staphylococcal enterotoxin B. The antibodies were all able to bind lactoperoxidase-labeled [125I]STa, and antibody 20B3 was also able to dissociate [125I]STa bound to toxin receptors on rat jejunal villous cells. Preincubation of STa with antibodies 20B3 or 20F5 led to a concentration-dependent neutralization of toxin activity in a suckling mouse intestinal secretion assay. These antibodies are likely to provide new tools for the continued study of STa structure-function relationships and may lead to improved diagnosis and treatment of E. coli-induced infectious diarrhea.
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PMID:Production of neutralizing monoclonal antibodies to Escherichia coli heat-stable enterotoxin. 388 Jul 23

The Mr 52,000 glycoprotein is regulated by estrogen and released by breast cancer cells in culture (B. Westley and H. Rochefort, Cell, 20: 352-362, 1980). This rare protein was partially purified from 25 liters of medium conditioned by MCF7 cells and injected into Biozzi's selected mice. The spleen lymphocytes of one immunized mouse was fused with the murine myeloma P3-X63-Ag8-653. Sixteen hybridomas producing monoclonal antibodies to the Mr 52,000 protein were isolated, and seven of them were cloned and purified. The seven monoclonal antibodies were all of the immunoglobulin G1 isotype, and their dissociation constants ranged from 0.35 to 2.3 nM. The antibodies specifically recognized the secreted Mr 52,000 protein as evidenced by double immunoprecipitation and by immunoblotting after electrophoretic separation and transfer. Double-determinant immunoradiometric assay indicated that the seven purified monoclonal antibodies recognized three distinct regions of the Mr 52,000 protein, and it was used to assay the Mr 52,000 protein in biological fluids. These antibodies did not react with the external plasma membrane of MCF7 cells, as shown by immunofluorescence analysis. By contrast, the cytoplasm of MCF7 cells (but not T47D and RBA cells) was stained by the peroxidase-immunoperoxidase complex after plasma membrane permeation, indicating that the protein is secreted by exocytosis rather than shed from the plasma membrane.
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PMID:Characterization of monoclonal antibodies to the estrogen-regulated Mr 52,000 glycoprotein and their use in MCF7 cells. 388 Nov 71


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