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Query: UMLS:C0026764 (multiple myeloma)
 36,148 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Monoclonal antibodies (MAbs) against HLA antigens can give better information about the serology and biochemistry of the human major histocompatibility complex (MHC) than HLA antisera obtained from pregnant women. To increase the very limited panel of MAbs against HLA we immunized mice with human cultured cell lines and fused their spleen cells with the Ag8-653 myeloma. We produced MAbs against HLA-A1, A2/w69, A2/28, A2/11/25/26/28/29/30/31/33/34, A25/32, B7/22, and B13. The best dilution medium to store the MAbs in Terasaki plates was RPMI 1640 supplemented with 7% bovine serum albumin. The MAbs are also excellent reagents to investigate public HLA antigens and to stain HLA antigens in cryosections of transplanted organs.
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PMID:Production and applications of new monoclonal antibodies against human lymphocyte antigen-A and -B antigens. 321 87

There have been many unsuccessful attempts to induce gametocytogenesis in vitro. In the present experiment, however, we found that RPMI-CS medium and RPMI-FS medium prepared by dissolving powdered RPMI 1640 medium in the culture supernatants of hybridoma cells, hybrid line D21 and 219.5, respectively, that produce anti-P. falciparum antibody induced gametocytogenesis. Gametocytogenesis was consistently observed from 3 days after addition of these media. The culture supernatant of anti-P. falciparum antibody producing hybridoma cells did not induce gametocytogenesis in the absence of RPMI 1640 medium. RPMI-MS medium, prepared by dissolving powdered RPMI 1640 medium in the culture supernatant of myeloma cells, SP2/O-Ag 14, which was used as a control, induced a few gametocytes.
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PMID:Induction of gametocytogenesis in Plasmodium falciparum by the culture supernatant of hybridoma cells producing anti-P. falciparum antibody. 330 78

Islet cell antibodies have been detected in more than 60% of newly diagnosed type I diabetics. Their pathogenetic role is still unclear. We have generated monoclonal antibodies (mc-ab) reactive with islet cell antigens by fusing mouse myeloma cells with spleen cells from Balb/c mice immunized with pancreatic islet cells. Hybridomas producing islet cell surface antibodies (ICSA) were detected by indirect immunofluorescence on viable cells from rat islets or rat insulinoma. Cytoplasmic islet cell antibodies (ICA) were detected by indirect immunofluorescence on Bouin-fixed sections of mouse pancreas. The ICSA- and/or ICA-producing hybridomas were cloned twice by limiting dilution. This paper describes six different mc-ab. All hybrid cell lines obtained produced IgM antibodies. Four of them mediate complement-dependent cytotoxicity to viable rat islet cells. In the present study the heterogeneity of circulating ICSA is demonstrated. Also, a monoclonal beta cell surface autoantibody K56aF3 was produced by fusion of spleen cells from a mouse treated with sub-diabetogenic doses of streptozotocin in combination with complete Freund's adjuvant. It was cytotoxic against islet cells up to a dilution of 1:1,000 and it could inhibit the insulin secretion from neonatal rat islets cultured in RPMI 1640 as stimulated by glucose or by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine common with glucose. The latter effect was reversible as indicated by the recovery of insulin secretion in a subsequent culture period without mc-ab. These results suggest that circulating ICSA in type I diabetics may alter beta cell function and thereby contribute to the pathogenesis of type I diabetes.
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PMID:Generation and partial characterization of monoclonal antibodies reactive with islet cell antigens. 331 74

The recently established culture medium, GIT, is applicable to many kinds of cells including mouse and human myeloma cells, and most adhesive cell lines. We applied this GIT medium to mouse B cell hybridoma production. When the medium was used to propagate myeloma cells before cell fusion and also for HAT selective medium, the fusion efficiency was more than twice as high as when the regular medium (RPMI-1640 supplemented with FBS) was used. Constantly more than 80% wells were hybridoma positive irrespective of the antigens used. To determine the optimal cell concentration at the hybridoma selection, a graded number of myeloma and spleen cells was distributed to each well; the best result was obtained when 3 X 10(4) myeloma and 3 X 10(5) spleen cells were distributed to each well. In addition, the GIT medium shows very little lot-to-lot variation. These results indicate that fusion efficiency in mouse B cell hybridoma production was greatly improved by using GIT medium.
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PMID:A great improvement of fusion efficiency in mouse B cell hybridoma production by use of the new culture medium, GIT. 331 8

Cytochemical and ultrastructural features of mouse hybridomas and also of the parental cells--myeloma P3-X63-Ag8.653 and spleen cells of the Balb/c mice immunized with cell line RPMI-1788 have been studied. Differences in cytomorphological signs and activity of acid phosphatase, acid nonspecific esterase, nonspecific-alpha-naphthyl acetate esterase were shown in hybrid cell lines secreting and not secreting monoclonal antibodies.
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PMID:[Morphocytochemical and electron microscopic research on murine hybridoma cells producing and not producing monoclonal antibodies]. 335 81

The glutathione (GSH) synthesis inhibitor, buthionine sulfoximine (BSO) was tested for cytotoxicity and thiol depletion in murine and human tumor cells in vitro, and for its antitumor activity and toxicity in vivo. The cell lines used in these studies included murine L-1210 leukemia, human RPMI 8226 myeloma, MCF-7 breast cancer and WiDr colon carcinoma. Soft agar colony forming assays showed that BSO was most effective at reducing tumor colony formation when exposed continuously to cells in vitro. Drug concentrations which inhibited colony formation to 50% of control levels ranged from 2.0-6.2 mM (for 1 hour exposures), 2-100 mM for 24 hour exposures and 0.4-1.40 microM (for continuous BSO exposures). Human myeloma cells proved most sensitive to BSO. In vitro cytotoxicity correlated with depletion of intracellular nonprotein sulfhydryls to less than or equal to 10% of control values in both L-1210 and 8226 cells. This was routinely achieved with prolonged exposures to mM BSO concentrations for greater than 24 hours. Normal mice tolerated high BSO doses (up to 5.0 g/kg) without evidence of acute toxicity. BSO was not active against L-1210 leukemia-bearing DBA/2 mice. When tested in vivo against MOPC-315 plasmacytoma-bearing BALB/c mice, BSO was not active at doses up to 4.0 g/kg. In contrast, the bifunctional alkylating agent melphalan (L-PAM) was active against MOPC-315 and this activity was enhanced by a 24 hour pretreatment of mice with 50 mg/kg of L-BSO.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Cytotoxic effects of glutathione synthesis inhibition by L-buthionine-(SR)-sulfoximine on human and murine tumor cells. 358 42

A serum-free medium was developed for culture of plasmacytomas and hybridomas that are dependent upon or independent of the presence of specific P388D1-derived polypeptide growth factors. Possibly due to the stringency of requirements for culturing such plasmacytomas, a highly advantageous combination of components was developed. The medium, SFM, contains RPMI 1640, beta-mercaptoethanol, Hepes buffer, reduced glutathione (GSH), selenite (Se), pyruvate, transferrin, albumin, soybean lipids, and low density lipoproteins. A cooperative effect of GSH and Se is observed. SFM supports the continuous, prodigious growth of every B cell line tested, including a variety of the plasmacytomas, hybridomas, myeloma fusion partners and lymphoma cell lines, as well as the macrophage-like cell line P388D1. All of the B cell lines grow equally as well in SFM as in serum-containing medium. Hybridoma lines from mouse-mouse and rat-mouse fusions continue to produce monoclonal antibodies which can then be purified in a single-step procedure. The cells can be cloned out and frozen down in SFM. In addition, the medium is highly effective for establishment of plasmacytoma cell lines from some transplantable plasmacytomas, including early generation tumors.
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PMID:Serum-free medium for growth factor-dependent and -independent plasmacytomas and hybridomas. 358 96

This report describes the formation of human hybridomas after in vitro immunization of peripheral blood lymphocytes (PBL) with an antigen and fusion of the stimulated lymphocytes with a HAT-sensitive human myeloma cell line, RPMI 8226. PBL were stimulated in vitro with sheep erythrocytes (SRBC) plus fresh human serum. PBL of some donors produced anti-SRBC antibody when they were cultured at 2 X 10(6) cells per well in a 24-well plate with the antigen plus fresh human serum for 7 days. Although lymphocytes of some donors were "low-responders" under the above conditions, they responded to SRBC when they were cultured with not only the antigen plus fresh human serum but also with the culture supernatant obtained after phytohemagglutinin (PHA) stimulation of a mixture of PBL from two donors (MLC-PHA sup). The cells sensitized by this procedure were fused with RPMI 8226 cells. Hybrids secreting IgM or IgG anti-SRBC antibodies were obtained. Additionally the ratio of total IgG-producing hybridomas to IgM-producing ones was higher when the MLC-PHA sup was used at the time of the in vitro immunization.
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PMID:Production of human--human hybridomas secreting antibody to sheep erythrocytes after in vitro immunization of peripheral blood lymphocytes. 361 Feb 34

Human CCRF-CEM ('T' cell), EB3p ('B' cell) and RPMI-8226 (myeloma cell) lymphocytic cell lines were an order of magnitude more sensitive to melphalan (MEL) than Chinese hamster, V-79-753B, cells even though the amount of [14C]MEL they incorporated was less than 50% of that incorporated into the rodent cells: the D0 values were 0.16, 0.20 and 0.30 microgram/ml respectively compared with 1.6 micrograms/ml. Furthermore, MEL sensitivity was not related to the total thiol content of the cells. DNA-DNA cross-linking was not detectable in lymphocytic cells using the alkaline elution technique at doses of MEL used for clonogenic survival, whereas in Chinese hamster cells both parameters were assessable within the same dose range. At high concentrations of MEL there was a direct relationship between DNA-DNA cross-linking and drug dose in each lymphocytic cell line. Changes in the amounts of DNA-DNA cross-linking, at different MEL concentrations, increased directly with the sensitivity of the cells, viz. CCRF-CEM greater than EB3p greater than RPMI-8226. Calculated survival values for doses of MEL which produced measurable DNA-DNA cross-linking showed that there was a similar relationship between these parameters for the three lymphocytic cell lines which was different from that for Chinese hamster cells. It is concluded that the contribution of DNA-DNA cross-links in determining cell survival after MEL treatment is both quantitatively and possibly qualitatively different in human and rodent cells and that DNA-DNA cross-linking cannot be used as an indicator of MEL sensitivity in human lymphocytic cells unless parallel clonogenic survival studies are also undertaken.
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PMID:Comparison of melphalan toxicity in human lymphocytic cells and Chinese hamster cells in vitro: the relationship between DNA-DNA cross-link formation and clonogenic survival. 362 62

The monoclonal antibody (MoAb) MM4 reacts with human multiple myeloma (MM) cell lines and bone marrow from patients with plasma cell dyscrasias but not with normal peripheral blood or bone marrow cells. Treatment with MM4 and rabbit complement (C') was cytotoxic to the plasma cell-derived cell lines GM 1312, RPMI 8226, and ARH-77, as demonstrated by chromium release microcytotoxicity and trypan blue exclusion assays. The same treatment eliminated greater than 99% of clonogenic myeloma stem cell colony formation of these cell lines, with less than 20% inhibition of normal human bone marrow pleuripotent progenitor colony formation in vitro. As an experimental model to explore the efficacy of MM4 + C' in purging MM-involved bone marrow, normal marrow cells were mixed with RPMI 8226 or GM 1312 cells in the ratio of 90:10 or 50:50 (marrow:myeloma cells). Colony growth assays indicated that MM4 + C' eliminated at least 2 logs of clonogenic myeloma stem cells in both 90:10 and 50:50 preparations, while sparing the majority of normal marrow progenitors (inhibition of CFU-C:10% to 13%; BFU-E:0%). The selectivity of MM4-mediated cytotoxicity may be useful for eliminating myeloma clonogenic stem cells from bone marrow of patients with multiple myeloma.
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PMID:Elimination of clonogenic stem cells from human multiple myeloma cell lines by a plasma cell-reactive monoclonal antibody and complement. 366 43


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