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)

Mouse hybridomas producing antibodies against structural proteins of canine distemper virus (CDV) were produced by fusion of Sp2/0 myeloma cells with spleen cells from BALB/c mice immunized with purified preparations of Vero cell-grown CDV. Ascites fluids collected after intraperitoneal inoculation with 149 CDV antibody-producing hybridoma cell lines were characterized by different serological tests. By immune precipitation tests with [35S]methionine-labelled extracellular virions and intracellular virus polypeptides, 57 clones were found to produce antibodies against the nucleocapsid protein (NP), 22 against the polymerase (P) protein, 10 against the fusion (F) protein and nine against the large uncleaved glycoprotein (named H in analogy with measles virus). By competitive binding enzyme-linked immunosorbent assay (ELISA) tests with monoclonal antibodies against each structural component, a minimum of 18, six, three and seven separate antigenic determinants were identified on the NP, P, F and H proteins, respectively. The reactions of clones directed against F and H surface components of the virus were tested for their ability to inhibit the infectivity of both CDV and measles virus in the absence and presence of anti-gamma-globulin. In addition, the inhibitory activity of the clones on measles haemagglutinating (HA) and haemolysis (HL) activity were examined. Monoclonal antibodies against six of the seven antigenic determinants of the H protein could neutralize the infectivity of the virus. After addition of anti-gamma-globulin to the test, increases of titres varying from twofold to several hundredfold were observed with the different clones. None of all the clones against H could block measles virus infectivity, HA or HL activity. The 10 clones directed against the F protein could not neutralize the infectivity of CDV even in the presence of anti-gamma-globulin. Further, the antibodies could not inhibit measles HA and HL activity in the absence of anti-gamma-globulin. However, after the addition of anti-gamma-globulin, antibodies against two of the three sites were found to block measles virus HL activity. The reactions of all clones were tested in immune fluorescence, ELISA and immune precipitation tests with three strains of CDV. Each strain had a few unique antigenic sites. Variation was found in four, one and three different antigenic sites of the NP, P and H proteins, respectively.
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PMID:Preparation and characterization of monoclonal antibodies directed against four structural components of canine distemper virus. 257 91

Cells producing neutralizing monoclonal antibodies to a serotype 3 human neonatal rotavirus strain RV-3 were derived by fusion of hyperimmunized mouse spleen cells with mouse myeloma cells. As ascites fluid, three rotavirus-neutralizing monoclonal antibodies were characterized by hemagglutination inhibition and reacted with 17 cultivable mammalian rotaviruses representing five virus serotypes, by fluorescent focus neutralization and enzyme immunoassay. Two antibodies, Mab RV-3:1 and Mab RV-3:2, reacted with the seven serotype 3 rotaviruses only. Mab RV-3:1 was shown to bind to the outer capsid glycoprotein gp34 of rotavirus when variants of SA 11 rotavirus were used, and it therefore appears to react with the major neutralization epitope of serotype 3 rotaviruses. The antibody Mab RV-3:3 was specific for an epitope of RV-3 rotavirus not present on any other rotavirus of any serotype tested, including another neonatal isolate of identical RNA electropherotype isolated from the same ward of the same hospital as RV-3 3 months earlier. These two viruses were also distinguishable by fluorescent focus neutralization, using antiserum to RV-3 virus. Western blot analysis showed binding of Mab RV-3:3 to the trypsin cleavage product of the outer capsid protein p86 of RV-3. This suggests that antigenic drift may have occurred among neonatal rotaviruses in Melbourne. These monoclonal antibodies will be useful in serotyping assays of rotaviruses directly in stool samples, and in further analysis of antigenic variation within the serotype.
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PMID:Neutralizing monoclonal antibodies to human rotavirus and indications of antigenic drift among strains from neonates. 257 49

By fusion of mouse NS1 myeloma cells with splenocytes from a BALB/c mouse immunized with human melanoma cells, an IgG1 monoclonal antibody, designated as 140.72, was produced. By the mixed hemadsorption antibody binding assay, 140.72 was shown to react with 17 of 20 melanoma cell lines and with 5 of 14 carcinoma cell lines. This antibody also reacted with 3 of 3 normal melanocyte cultures in much lower titers. It did not react with any of 35 other normal and malignant lines, including neuroblastoma, glioblastoma, sarcoma, teratoma, fibroblast, and lymphoid cell lines. Absorption with fresh melanoma and carcinoma homogenates confirmed the results of direct tests. Fetal reactivity of antibody 140.72 was determined by positive absorption with 10 of 11 tissue homogenates derived from different fetuses of 10-16 weeks' gestation. The reactivity of this antibody was completely removed by absorption with a highly purified preparation of carcinoembryonic antigen (CEA) derived from a colon carcinoma. The antigenic activity was detected in the culture medium of reactive cell lines. Immunoprecipitation analyses of melanoma and carcinoma cells indicated that the antigenic determinant recognized by antibody 140.72 is on a glycoprotein with an apparent molecular weight of 95,000-150,000 common to both serologically reactive cell types. Additionally, a 200,000-molecular-weight glycoprotein corresponding to the CEA molecule was detected only on the reactive carcinoma cells. These data confirmed previous findings obtained with polyclonal anti-CEA antisera for the existence of shared CEA-related antigenic determinants on human carcinomas and melanomas and provided additional molecular characterization of these glycoproteins. Further characterization of the molecules bearing the antigenic determinant recognized by antibody 140.72 should be performed with a view to exploring its potential in the immunodiagnosis and immunotherapy of patients with melanoma.
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PMID:Monoclonal antibody recognizing human melanoma-carcinoma cross-reacting oncofetal antigen epitopically associated with carcinoembryonic antigen. 258 73

Monoclonal antibodies (MAbs), for human use require chemical and biological purity. The best approach seems in vitro cultivation in a serum-, protein-free medium. A basal defined culture medium has been developed to sustain optimal hybridoma cell growth and MAb secretion. It consists of Iscove's Dulbecco's modified, Eagle's, Ham's F12 and NCTC 135 media in a 5:5:1 mixture (v/v/v), to which glucose is added to reach a final concentration of 25 mM, glutamine to 4-6 mM, 2-mercaptoethanol to 50 microM, Pluronic F68 to 0.01-0.1% (w/v), Hepes to 25 mM and NaHCO3 to 3 g/l. Hybridoma cells, derived from Sp 2/0 myeloma and secreting a MAb to a human milk fat globule membrane-associated high molecular weight glycoprotein, were cloned in this medium containing 1% (v/v) fetal calf serum and then sequentially adapted in serum-free medium further supplemented with transferrin and insulin, both at 10 micrograms/ml. Clones producing immunoreactive MAbs secrete a mean of 50 micrograms IgG/ml, i.e., ca. 80% of the concentration reached in Dulbecco's modified Eagle's medium containing 10% serum. When cells were cultured in spinner flasks with a semi-continuous mode of cultivation (with a daily removal of 20% of the volume and its replacement by fresh culture medium), in serum-free medium further supplemented with 10 nM estradiol, a mixture of trace elements and albumin (at 30 micrograms/ml) complexed to linoleic acid, MAb secretion reached 100 micrograms/ml and became equal or higher to that obtained in serum-containing medium. MAb secretion was not decreased and was even significantly increased during the growth phase, when transferrin was replaced by another iron source, i.e., ferric citrate at 500 microM associated with 20 microM ascorbic acid. Finally, deletion of insulin and of albumin-linoleic acid did not affect significantly cell density nor MAb secretion. In conclusion, it appears from this study that semi-continuous cultivation in spinner flasks of hybridoma cells, after cloning and progressive adaptation, in a chemically defined, serum- and protein-free medium, permitted MAb secretion to be increased to a mean of 144 micrograms/ml, i.e., multiplied by a factor of ca. 1.5 compared to culture of these cells in serum-containing medium under the same conditions and by a factor of ca. 2.4 compared to cultivation in serum-containing medium in flasks.
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PMID:Optimisation of hybridoma cell growth and monoclonal antibody secretion in a chemically defined, serum- and protein-free culture medium. 264 56

Experiments to determine the hydrolysis and protein binding of melphalan (L-phenylalanine mustard, L-PAM) were carried out in vitro for therapeutic concentration of the drug: the decrease in L-PAM concentration in plasma and whole blood during 24 h incubation at 37 degrees C was only 5% due to hydrolysis. Serum protein binding was about 90%, whereby 60% and 20% of this binding was due to interactions with albumin and acid alpha 1-glycoprotein, respectively. Immunoglobulins did not participate in the binding of L-PAM. The covalently bound part of L-PAM in serum was 30% in the concentration range of 1-30 micrograms/ml. The binding of dihydroxymelphalan (DOH) in serum did not exceed 20%. Glucocorticoids used in combination with L-PAM for treating multiple myeloma did not influence its protein binding. Our study with 35 sera from 15 patients with multiple myeloma shows that high levels of paraproteins do not increase but may decrease the binding of L-PAM, resulting in an elevated concentration of free drug.
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PMID:Relevance of the hydrolysis and protein binding of melphalan to the treatment of multiple myeloma. 291 May 15

Mouse and rat myeloma cell lines showed little or no rosette formation with sheep erythrocytes (SRBC) coated with rabbit IgG (rabbit EA) but showed marked rosette formation after treatment with trypsin, pronase or neuraminidase. These cell lines showed no rosette formation with SRBC coated with mouse IgG (mouse EA): treatment with trypsin enabled the detection of rosettes among the mouse myeloma cell lines but not by the rat myeloma cells. The F(ab')2 fragment of the anti-Fc receptor II antibody blocked the formation of rosettes with rabbit EA by mouse myeloma cell lines after treatment with trypsin. Aggregated mouse IgG1 and IgG2b subclasses strongly inhibited the formation of rosettes with rabbit EA, whereas aggregated mouse IgG2a showed a marginal inhibitory effect. A large amount of mouse IgG2a, however, caused significant inhibition. Our results also revealed that aggregated mouse IgG could bind to the rat myeloma cell line. The Fc rosette forming abilities of the enzyme-treated mouse and rat myeloma cells became reduced after cultivation both in the presence and absence of FCS but not after cultivation in the presence of cycloheximide, suggesting that cell surface substances, which may be glycoprotein that incompletely mask Fc receptors, are produced by myeloma cells.
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PMID:Studies on the Fc receptors and masking substances on mouse and rat myeloma cells. 297 95

Hybridomas producing human monoclonal antibodies (MAbs) against herpes simplex virus (HSV) were established by fusing human tonsillar lymphocytes with mouse myeloma cells. Three hybridomas have been stably producing MAbs for more than 16 months. All three MAbs--H1, H2, and H3--were of the IgG1 isotype and recognized the gB glycoprotein of HSV types 1 and 2 (HSV-1 and HSV-2). MAbs H2 and H3 not only bound to the surface membrane of HSV-infected cells but also neutralized both HSV-1 and HSV-2, whereas MAb H1 had neither activity. In mouse infection experiments, MAbs H2 and H3 showed a potent protective effect against HSV-1 infection, whereas MAB H1 was less protective. Furthermore, the development of zosteriform skin lesions in athymic nude mice was suppressed by administering MAb H2. These results suggest that human MAbs might provide passive immunization against HSV infections in humans.
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PMID:Antiviral activities of human monoclonal antibodies to herpes simplex virus. 302 8

Hybridomas producing human monoclonal antibodies (mAb) against varicella-zoster virus (VZV) were generated by fusing human splenic lymphocytes with mouse myeloma cells. Before cell fusion, lymphocytes were stimulated in vitro with viral antigens and pokeweed mitogen. This combination synergistically increased the generation of VZV-specific hybridomas. Five established hybridomas have been stably producing mAb for at least 9 months. These mAb, designated V1, V2, V6, V8 and V9, were of the IgG1, lambda isotype. They bound to all 6 tested VZV strains but not to other herpes viruses, with the exception that V1 bound to herpes simplex virus (HSV) as well as VZV. Immunoprecipitation analysis showed that V1, V6 and V9 recognized glycoprotein gpII, whereas V2 and V8 recognized gpI. In addition, V1 reacted with the gB glycoprotein of HSV. All these mAb neutralized viral infectivity. The neutralizations by V2 and V8 were more effective and more complement dependent than those by V1, V6 and V9. Immunofluorescence tests revealed that all these mAb bound to the surface membrane of VZV-infected cells. These results suggest that cell fusion between in vitro stimulated lymphocytes and mouse myeloma cells is a reliable method for the generation of hybridomas capable of stable production of human mAb. The human mAb thus developed may provide a new means of passive immunization of humans against VZV infection.
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PMID:Hybridomas producing human monoclonal antibodies against varicella-zoster virus. 303 47

Two neutralizing monoclonal hybridoma antibodies to the surface spike glycoprotein E2 of the JHM strain of murine hepatitis virus were injected into homologous BALB/c mice, and their biological half-lives were determined by sequential titration of plasma samples in a virus-specific enzyme immunoassay. Intravascular half-lives of monomeric immunoglobulins were estimated at 8.0 +/- 1.5 days for antibody 5B19.3, an IgG1, and 12.7 +/- 2.4 days for antibody 4B11.6, an IgG2a. These catabolic rates are statistically different from each other (P less than 0.001) and significantly higher than previously reported values, which were all obtained with radiolabelled polyclonal or myeloma immunoglobulins of unknown specificities. Failure to remove aggregated 4B11.6 antibodies by high-speed centrifugation yielded a statistically significant acceleration of biological turnover (half-life 9.9 +/- 1.6 days; P less than 0.01).
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PMID:Catabolism of homologous murine monoclonal hybridoma IgG antibodies in mice. 303 67

A novel cell surface marker of fetal development was identified in both in vivo and in vitro systems of the mouse using monoclonal antibodies against a glycoprotein of an apparent size of 133,000 Da. Two independent clones of hybridomas were isolated by fusing murine myeloma cells, NS-1, with spleen cells of a rat which was immunized with murine 3T3 fibroblast. The analysis of molecular size and tryptic peptides of the immunoprecipitate indicated that fibroblast and putative parietal endoderm cells, which were derived by induced differentiation of F9 embryonal carcinoma cells with retinoic acid and cyclic AMP, expressed apparently the same protein. Undifferentiated F9 cells and F9 cells which were treated with retinoic acid or cyclic AMP alone had little or no immunoprecipitable proteins. Analogously, parietal endoderm of in vivo embryos tested positive for this protein but visceral endoderm and embryonic ectoderm did not. The amount of this surface protein was increased in fibroblast and differentiated F9 cells by elevation of intracellular cyclic AMP concentrations. These results are consonant with a hypothesis that this surface protein plays a role in fetal development via a quantitative modulation by cyclic AMP.
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PMID:Fetomodulin: marker surface protein of fetal development which is modulatable by cyclic AMP. 303 32


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