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

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

A human-mouse myeloma analogue termed HMMA2.11TG/O was constructed by fusion of the mouse myeloma cell line P3x63Ag8.653, a mutant derivative of MOPC21, with bone marrow mononuclear cells from a patients with IgA myeloma. The HMMA2.11TG/O cell line is resistant to 6-thioguanine and ouabain and sensitive to HAT. The cell line secretes no detectable immunoglobulin and has a hybrid karyotype and cell surface phenotype. An average fusion efficiency for growth of hybridomas of 1/17,000 fused cells was obtained in fusions with human peripheral blood mononuclear cells (PBM), Pokeweed Mitogen (PWM) stimulated PBM, and Epstein-Barr Virus (EBV) transformed polyclonal B cell lines. Over 75% of hybrids secrete detectable immunoglobulin and the cloning efficiency of the hybrids at 1 cell/well averages 25%. Antibody secreting cloned hybridoma cell lines were obtained by fusion directly with PBM from an immunized volunteer and by fusion with in vitro, secondarily immunized, EBV transformed polyclonal cell lines. Five hybridomas secreting human monoclonal IgM anti-tetanus antibodies and 2 secreting human monoclonal IgG anti-tetanus antibodies were selected and cloned from 6 fusions performed specifically for anti-tetanus antibody. Immunoglobulin and antibody secretion by cloned hybrids has been stable for 5-10 months at present. Immunoglobulin and antibody secretion in routine cultures passaged every 3-4 days has been 8-42 micrograms/ml. This human-mouse myeloma analogue should prove useful for the routine production of human monoclonal antibodies.
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PMID:The construction and use of a human-mouse myeloma analogue suitable for the routine production of hybridomas secreting human monoclonal antibodies. 343 25

Autoantibody-secreting hybrid cell lines were obtained by fusion of spleen cells from unimmunized (NZB X NZW) F1 mice with the HAT-sensitive mouse myeloma cell line SP2/0-Ag14. Eight hybridoma cell lines producing autoantibodies to mouse thymocytes were cloned and the resultant antibodies were partially characterized. All eight monoclonal antibodies lysed mouse thymocytes in the presence of rabbit complement. The anti-thymocyte cytotoxic antibody activities were absorbed with thymocytes, lymph node cells, unfractionated spleen cells, and splenic T cells; but not with bone marrow cells, splenic B cells, or homogenates of mouse kidney, liver, or striated muscle cells. In addition, the cytotoxic activities of culture supernatants from seven of the eight hybrid clones were absorbed with mouse brain tissue homogenates. Isotyping of the monoclonal antibodies revealed that five were IgM and three were IgG2a. Mouse thymocytes sensitized with each of the eight monoclonal antibodies in vitro became highly susceptible to phagocytosis by syngeneic macrophages. The monoclonal antithymocyte antibodies, thus, appear to be similar to the naturally occurring, (NZB X NZW) F1 thymocytotoxic autoantibodies (NTA) described by Shirai et al.
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PMID:Establishment of monoclonal antibodies which possess the same characteristics as the naturally occurring thymocytotoxic autoantibodies (NTA). 349 9

We have established a new human myeloma cell line from the pleural effusion of a patient with an IgA lambda myeloma, using special tissue culture conditions and selection procedures to prevent the outgrowth of contaminating Epstein-Barr virus (EBV)-carrying normal B-lymphoblastoid cells present in the explant. The myeloma cell line, U-2030, is aneuploid and EBNA-negative and has morphological features, reactivity with cytochemical markers and cell-surface antigen expression typical of plasmablasts. The cell line thus appears to be representative of the malignant clone in vivo. However, functionally the line is a non-Ig-producer and must therefore be derived from a non-secretory variant cell present within the highly aneuploid myeloma cell clone in vivo. The U-2030 differs from previously established human myeloma cell lines in that it has a comparatively high growth rate, is clonable and can be made HAT-sensitive relatively easily. This, together with the facts that it is a non-Ig-producer and mycoplasma-free, suggests that the 6-thioguanine-resistant, HAT-sensitive subline, U-2030 TG, derived from this cell line may be used as a malignant fusion partner for the production of human-human hybridomas. An EBV-carrying lymphoblastoid cell line (U-2031) was also established. This line was diploid and had all the phenotypic properties of lymphoblastoid lines established from normal individuals.
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PMID:Establishment of a new human myeloma cell line (U-2030) and selection of a hat-sensitive subline. 358 53

In attempts to produce stable lines secreting bovine monoclonal antibodies, murine/bovine hybridomas (1 degree xenohybridomas) were selectively cultured in 8-azaguanine to derive HAT-sensitive lines that were then used as myeloma partners for further fusions with bovine lymphocytes. The resulting 2 degrees xenohybridomas were further selected to produce 3 degrees xenohybridomas. Four stable lines secreting bovine monoclonal antibodies recognizing blood group determinants X1 (an IgG1), E'2 (an IgM) and SU" (an IgGI) and another (an IgGI) as yet unidentified were produced from fusions of 2 degrees xenohybridomas with lymphocytes from calves that had been immunized with bovine red cells.
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PMID:Murine/bovine hybridomas producing monoclonal alloantibodies to bovine red cell antigens. 360 87

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

The therapeutic efficacy of antineoplastic purine analogs can be jeopardized by the emergence of drug-resistant mutant subpopulations of tumor cells. To determine whether such mutant populations might be eradicable in vivo with the type of HAT combination (hypoxanthine + an antifolate + thymidine) known to be selectively cytotoxic to them in vitro, 2 thioguanine-resistant BALB/c murine myeloma lines were transplanted into BALB/c mice to produce tumors capable of progressive growth in the absence of therapy. Treatment of these mice with a modified HAT regimen induced permanent tumor regressions in 37/44 mice; the same treatment was ineffective against tumors produced by a non-mutant myeloma line from which one of the mutant sublines had been derived.
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PMID:In vivo efficacy of HAT therapy against transplantable murine myelomas resistant to purine analogs. 371 70

Mouse myeloma NS1-Ag4 cells were fused with spleen cells from a BALB/c mouse previously immunized with luteinizing hormone releasing hormone (LHRH) conjugated to serum bovine albumin (BSA). Fused cells were grown in HAT restrictive medium which was screened for LHRH binding ability via a primary binding assay employing [125I]LHRH and cold ethanol precipitation. One clone (hy-USASK/DSIL-LHRH-A1) was selected for further study. Cell culture fluid and ascites fluid bound 30% of [125I]LHRH at 1:4000 and 1:400,000 dilution respectively. A competitive inhibition assay using ascites fluid at 1:2,000,000 dilution and LHRH standards at 0.125-32.0 ng/ml was established. Initial studies using rabbit anti-mouse allotype sera in a horseradish peroxidase (HRP)-ELISA system indicate the antibody is IgG1. A dose of 0.5 ml ascites fluid containing LHRH antibody given intravenously (i.v.) on day 9 of gestation was effective in terminating pregnancy in rats. A 1 cm progesterone implant made of elastomer polymer and placed interperitoneally blocked this effect. Ascites fluid (4.5 ml) containing LHRH antibody, when infused i.v. into mature spayed female dogs induced a precipitous decline in mean luteinizing hormone (LH) levels and reduced LH pulsatility over 4 days. It was concluded that the mouse monoclonal antibody is specific for LHRH, and can interrupt reproductive events in vivo.
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PMID:Monoclonal antibodies against LHRH: development and immunoactivity in vivo and in vitro. 388 2

In experiments aimed at generating monoclonal antibodies against tumour associated antigens, mice are usually immunized with cancer cells (or membrane fractions prepared from it) obtained from surgery or cultured in vitro. In both situations there is the danger of introducing mycoplasmas into the hybridoma cultures even if the myeloma cell line and the mice used for immunization or feeder cell preparation are not infected. Mycoplasmas kill hybridoma cells during HAT selection because they excessively degrade thymidine. The use of azaserine instead of aminopterin in hybrid selection circumvents the necessity for thymidine and therefore may allow survival and growth of hybridomas in the presence of mycoplasmas.
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PMID:Monoclonal antibodies against tumour-associated antigens: mycoplasma as a major technical obstacle and its possible circumvention by azaserine selection medium. 406 91

We have isolated three clones of hybrid cells which synthesize antibodies specific for determinants on simian virus 40 (SV40) T antigens. Mouse myeloma NS1 cells were fused with spleen cells from mice that had been immunized with SV40-transformed mouse cells. Hybrid cells were selected in HAT medium and cloned in soft agar. We used an enzyme-linked immunosorbent assay for detection and quantification of mouse antibodies against SV40 T antigens. Monoclonal antibodies from 3 of the 24 clones that scored as positive in the enzyme-linked immunosorbent assay were verified by immunoprecipitation to be specific for SV40 T antigens. Two clones (7 and 412) produced antibodies that recognized denaturation-sensitive antigenic determinants unique to large T antigen. Antibodies from clone 7 appeared to have a low affinity for large T antigen. Antibodies from clone 412 had a higher affinity for large T antigen but did not recognize a subclass of large T antigen that was recognized by tumor serum. Antibodies of the third clone, clone 122, recognized a denaturation-stable antigenic determinant of the 53,000-dalton mouse nonviral T antigen in SV40-transformed cells. Antibodies from clone 122 also recognized similar (51,000- to 56,000-dalton) nonviral T antigens in SV40-transormed or lytically infected cells from five mammalian species and in four uninfected mouse lines. From these observations, we have concluded that (i) the 94,000-dalton SV40 large T antigen may exist as immunologically distinguishable subclasses, and (ii) the nonviral T antigens of five mammalian species share at least one antigenic determinant.
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PMID:Monoclonal antibodies against simian virus 40 T antigens: evidence for distinct sublcasses of large T antigen and for similarities among nonviral T antigens. 615 77


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