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)

The HindII fragment of human interferon beta (IFN-beta) gene was inserted downstream from SV40 late promoter in pSV2-dhfr and cotransfered with pSV2-gpt into the mouse myeloma SP2/0 cells which were hypoxanthine guanine phosphoribosyl transferase (HGPRT) deficient. After the selection in HAT (Hypoxanthine Aminopterine Thymidine) medium containing myciphenolic acid and xanthine, the efficient constitutive expression of IFN-beta could be detected in the supernatant of the survive cells.
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PMID:[Expression of human interferon beta gene controlled by SV40 late promoter in mouse myeloma SP2/0 cells]. 248 40

Four monoclonal IgG antibodies to purified, recombinant murine gamma-interferon (rIFN-gamma) have been produced by fusion of immune hamster splenocytes with HAT-sensitive murine myeloma cells. Specificity was confirmed either with an enzyme-linked immunosorbent assay (ELISA) that used immobilized rIFN-gamma or with a radioimmunoassay that employed soluble 125I-rIFN-gamma and heat-killed, fixed Staphylococcus aureus-bearing Protein A. Competition binding experiments suggested that the monoclonal antibodies (MoAb) displayed two distinct epitope specificities: one displayed by H1 and H2, and the other displayed by H21 and H22. By using murine-human recombinant IFN-gamma hybrid molecules, the H1/H2 epitope was shown to depend on the amino-terminus of IFN-gamma, whereas the H21/H22 epitope was formed by the carboxy-terminal amino acid sequence. The MoAb also reacted with natural IFN-gamma. When bound to a surface, all four MoAb, but not normal hamster IgG, removed 100% of the antiviral and MAF activities present in supernatants of cultures of the murine 24/G1 T cell hybridoma. In free solution, all four antibodies inhibited IFN-gamma dependent antiviral activity, but with different efficiencies. Soluble H21/H22 also blocked all of the 24/G1-derived activity that induces nonspecific tumoricidal activity in macrophages (MAF) while H1/H2 enhanced MAF activity. The differential inhibitory or enhancing activities of H21 or H1 reflected their ability to inhibit or enhance binding of 125I-rIFN-gamma to macrophages, respectively. Soluble H21/H22 and solid-phase H1/H2 inhibited 100% of the MAF, microbicidal, and Ia-inducing activities from lymphokine preparations produced by mitogen stimulation of normal murine splenic cells. These results help to establish definitive structure-function relationships for the IFN-gamma molecule, and indicate that IFN-gamma is the primary lymphokine responsible for inducing nonspecific tumoricidal activity and Ia antigen expression, and for enhancing microbicidal activity in macrophages.
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PMID:Monoclonal antibodies to murine gamma-interferon which differentially modulate macrophage activation and antiviral activity. 257 13

Cells from adult Fasciola hepatica were fused with cells from a murine BALB/c myeloma Sp2 line. The hybrid cells were grown in HAT (hypoxanthine, aminopterin, and thymidine) medium, cloned and subcloned, and shown to express parasite antigen for 1 year after fusion. Expression of parasite antigen was demonstrated by the following: 2 histogram flow cytometric analyses, in which a population of hybrid cells in the population of 7 month cultured hybrid cells showed 57% more fluorescence when treated with an anti-F. hepatica serum followed by anti-rabbit immunoglobulin G coupled to fluorescein isothiocyanate as compared with the same hybrid cells washed and treated with normal rabbit serum; Sp2 myeloma cells treated with an anti-F. hepatica serum or normal rabbit serum followed by fluorescein-labeled anti-rabbit IgG had the same negative fluorescence; BALB/c mice immunized with PBS-washed cells from a subclone of these hybridomas developed anti-F. hepatica antibodies (shown by the Falcon assay screening test enzyme-linked immunosorbent assay); and antibodies recognized an F. hepatica antigenic polypeptide of 57,000 Mr in a Western immunoblot. These helminth:myeloma hybrids expressed murine host markers, further confirming the hybrid nature of this cell line. F. hepatica cells alone, like their Sp2 fusion partners, die in HAT supplemented medium by 9 days of culture. F. hepatica:Sp2 hybridomas have been grown continuously in HAT medium for greater than 1 year.
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PMID:Fasciola hepatica:Sp2/0 (helminth:myeloma) hybridoma expressing parasite antigen. 264 45

We have used the cholesterol auxotrophy of NS-1 mouse myeloma cells as the basis for selecting NS-1 hybridomas. The outgrowth of nascent NS-1 hybridomas in cholesterol-free serum-free medium was 3- to 9-fold more efficient than that in HAT medium and resulted in 3- to 13-times as many antigen-reactive hybridoma wells. This method of hybridoma selection can be applied with any sterol-dependent parent cell line. Hybridomas established under serum-free culture conditions were growth inhibited by fetal calf serum.
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PMID:An alternative method for the isolation of NS-1 hybridomas using cholesterol auxotrophy of NS-1 mouse myeloma cells. 273 3

A method of producing human monoclonal antibody by combining somatic cell hybridization technology with the capability of Epstein-Barr virus (EBV) to transform human B lymphocytes is described. Peripheral blood lymphocytes from a donor with positive tuberculin skin test reaction were transformed by EBV and then tested for antibody production to mycobacterial purified protein derivative (PPD) by an enzyme-linked immunosorbent assay. Two EBV-transformed lymphoblastoid cell lines making IgM antibodies to PPD were obtained. One of these cell lines was fused by polyethylene glycol with a murine hypoxanthine-guanine phosphoribosyl transferase-deficient myeloma cell line that had been selected for resistance to ouabain. The human-mouse hybrids were selected in ouabain-containing HAT medium and 11 heterohybridomas producing IgM antibody to PPD were obtained. One of these was cloned by limiting dilution with efficiency at least 20-fold higher than parent EBV-transformed cell line. Heterohybridoma subclones reached levels of IgM antibody as high as 75.0 micrograms/ml of culture medium, whereas IgM production of EBV-transformed B cell clones ranged between 3.0 and 4.0 micrograms/ml.
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PMID:Human monoclonal antibody to purified protein derivative of tuberculin produced by hybrids constructed with Epstein-Barr virus-transformed B lymphocytes and mouse myeloma cells. 300 5

Human monoclonal antibodies specific for the Rh(D) antigen were produced by cell lines generated by the fusion of pooled Epstein-Barr virus (EBV)-transformed B-cell lines secreting Rh(D) antibodies with the murine myeloma cell line NS.1 or with the human lymphoblastoid cell line HOA.1. The selection of hybrids was achieved in RPMI 1640 medium containing HAT and ouabain. Higher fusion efficiency was obtained with the NS.1 cell line; however, the hybrids with HOA.1 exhibited a greater clonal stability. The products of four clones (three human-human and one human-mouse) that consistently secreted antibodies for over 11 months were tested for specificity with a panel of red cells of various Rh phenotypes. The supernatants of all four clones showed anti-Rh(D) specificity but failed to react with the red cell Du phenotypes categorized as DV(Dw+) and DVI. Two of the three human-human clones secreted IgM(lambda) and the third IgG(kappa). The human-mouse clone produced IgG(kappa) antibody.
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PMID:Production and characterization of human-human and human-mouse hybridomas secreting Rh(D)-specific monoclonal antibodies. 303 6

The secretion of immunoglobulin (Ig) from cultured mononuclear cells by lipopolysaccharide (LPS) stimulation is inhibited by monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by murine T cell hybridoma. In an attempt to develop a murine monoclonal antibody (MAb) with specific reactivity against MNSF, a cell fusion technique that incorporated immune murine splenocytes and HAT-sensitive murine myeloma cells was used. Cross-reactivity experiments confirmed that the MAb (MO6) does not bind to unrelated proteins such as bovine serum albumin, mouse IgG, and murine interferon-gamma (IFN-gamma). There are no effects when anti-IFN-gamma antibodies are used with MNSF. As far as biological activity is concerned, MO6 inhibits in vitro the activity of MNSF in terms of the Ig secretion from cultured lymphocytes. By using MO6, affinity chromatography and immunoblotting were performed. The MNSF on the SDS-PAGE showed a band with m.w. of approximately 70,000, indicating the formation of an aggregate in saline; but after treatment with 0.4 M pyridine-acetic acid buffer, separate bands of 24,000 and 16,000 daltons were evident. Therefore MO6 recognizes 70,000 and both 24,000 and 16,000 daltons. Thus we confirmed by using this MAb and affinity chromatography, the existence of human counterpart, human nonspecific suppressor factor (hNSF), in supernatant from concanavalin A-stimulated T cells. When hNSF was fractionated by high pressure liquid chromatography (HPLC), the activity was found in a region corresponding to 70,000 daltons. However, when fractionated in pyridine-acetic acid buffer, hNSF activity was distributed in a slightly wider range of 15,000 to 30,000 daltons. Physicochemical analysis showed that the purified hNSF is resistant to either heating at 56 degrees C or to 2-mercaptoethanol treatment; however, it is labile to acidification at pH 2.0 and is also sensitive to protease treatment, the characteristics of which were similar to those of murine MNSF. Thus MO6 was confirmed to be a pertinent tool for isolation of hNSF, as well as for murine MNSF.
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PMID:Characterization of monoclonal nonspecific suppressor factor (MNSF) with the use of a monoclonal antibody. 310

A clone of cells secreting an antibody to an epidermal antigen was generated from a patient with a blistering skin lesion. Although produced by fusion of human lymphocytes to a HAT-sensitive myeloma, this clone of cells did not have characteristics of a hybridoma. A true hybridoma was produced by fusion of this clone to a HATr/ouabain(r) myeloma line. The IgM antibody secreted by this clone reacted with the intercellular region of the epidermis of normal human skin in a manner similar to pemphigus autoantibodies. In addition, in normal human kidney the antibody bound to glomeruli and tubules. It also reacted with an antigen present in the cytoplasm of a wide variety of cell lines including epithelial, lymphoid and myeloid types. No reaction was found with the surface of any of the cell lines, nor with DNA or phospholipid antigens. This monoclonal antibody may define an autoantibody specificity which mediates some autoimmune skin lesions. Its polyspecificity is reminiscent of some other human hybridoma autoantibodies, and its reaction with components of the kidney suggests an alternative pathology for renal disease in such patients.
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PMID:Production of a polyspecific human monoclonal antibody reacting with an epidermal antigen. 315 60

Several factors which affect the production of murine hybridomas by electrofusion using SP2/0 myelomas were examined. These factors included the ionic composition, specific resistance and osmotic strength of the fusion medium, proteolytic pretreatment of the cells, and field strength of the single, rectangular, 15 or 20 microseconds electric pulse. Individual experiments were observed by phase microscopy, video-recorded, and subsequently analyzed. The efficiency of hybridoma formation was measured by the number of hybrid colonies which survived HAT medium selection. In most cases, the peak efficiency occurred when the pulse field was between 3 and 4 kV/cm. Ca++ and Mg++, in 0.1 and 0.5 mM concentrations, respectively, helped to prevent large-scale SP2/0 lysis following pulse application. Reduction of the osmolarity of the fusion medium allowed for an approximate two-fold increase in the hybrid production efficiency. However, other variations, including an increase of the specific resistance of the medium to 1.7 x 10(4) omega cm, and a reduction of the pulse width to 15 microseconds, allowed significantly higher efficiencies. Dispase pretreatment of the myelomas led to additional improvements when using a 15 microseconds pulse. The data suggest that for this system, the highest efficiencies are obtained by maximizing the attractive polarization forces between cells, while adjusting the ionic composition and pulse parameters to preserve myeloma viability.
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PMID:Optimization of electrofusion parameters for efficient production of murine hybridomas. 319 35

Using the human liver cancer DNA transfected NIH/3T3 cell line, the human N-ras oncogene and the over expression of the oncoprotein P21ras was demonstrated, BALB/C mice were immunized. The spleen cells from the immunized mice were fused with SP2/0 myeloma cells. After the HAT medium selection and screening, two hybridoma cell lines, SCI-Oncogema 1 and 2, were established. In the immunoprecipitation test, the molecular weight of the protein reacting to Oncogema 1 was 21,000. This M.W 21,000 protein possessed the capability to bind with GTP, i.e. the character of P21ras. These data indicate that the Oncogema 1 is the monoclonal antibody against P21ras. Using Oncogema 1, specimens from 6 liver cancer patients were studied by immunopathology. With ABC stain, it was observed that the malignant cells in all the samples showed dark staining; the P21ras revealed over expression. Although the staining was heterogeneous, it implied that the ras oncogene was involved in the carcinogenesis of these six samples. No over expression was seen in the normal liver cells even in those around the cancerous lesion. However, dysplastic cells were moderately stained which means that the ras oncogene was activated and P21ras over expressed in these cells. The results suggest that the ras oncogene and P21ras play an important role in the early stage of liver cancer carcinogenesis.
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PMID:[Localization of oncoprotein P21ras in the human liver cancer]. 330 83


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