摘要:

Small-cell carcinoma (SCCL) is an aggressive type of lung cancer. Though it is usually responsive to therapy, be it chemotherapy or radiation, the majority of patients eventually relapse and overall prognosis is dismal. New forms of therapy are, therefore, needed. SM1 monoclonal antibody (MAb) was developed in our laboratory and demonstrated to be highly reactive against SCCL. I125radiolabeled SM1 antibody was administered intravenously to nude mice bearing SCCL tumor xenografts. The mice were sacrificed, different tissues sampled and tested for uptake of radioactivity five days following antibody injection.There was over a 30 fold increase in localization of labeled antibody to the tumor as compared to muscle tissue. All organs tested showed an insignificant amount of MAb (p=0.01) including the spleen, which had the highest normal tissue uptake in these experiments. These results demonstrate that SM1 MAb can be successfully targeted to SCCL xenografts. Its potential applications for imaging and therapy of SCCL in man are currently under investigation.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.1

年代:1988

数据来源: MAL

摘要:

First cycle transformants of NIH 3T3 cells transfected with metastatic human thyroid carcinoma DNA were used as immunogen to obtain monoclonal antibodies (MAbs) against normal and transformation-related antigens. The transformed cell line (M33) was shown to contain Alu sequences. Two MAbs were selected on the basis of their differential reactivity toward untreated NIH 3T3 cells or the transformed M33 cell line. By immunofluorescence, immunoelectronmicroscopy and biochemical analysis, the first MAb (MTr1) was demonstrated to recognize an epitope on cytoskeletal filaments of proliferating murine fibroblasts. Similar MTr1-labelled filaments were also found to accumulate into cytoplast-like structures spontaneously produced by M33 cells.The characterization by immunofluorescence of MTr2, the second MAb, indicates that it recognizes a specific human antigen associated with normal thyroid epithelial cells and differentiated thyroid tumors.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.7

年代:1988

数据来源: MAL

摘要:

Eight distinct murine monoclonal antibodies that are directed to normal human mouse small intestinal epithelial cells have been characterized. Three of these called TPNG 2, 20 and 43 were directed to goblet cells and mucin components. One antibody, TPNC 10, was directed to basilar crypt cell inclusions and was specific for the small bowel and possible progenitor cells. Another antibody, TPNC 30, was directed to the microvilli of the brush border. Finally, three antibodies, TPNC 50, 51 and 52 reacted with epithelial cell membranes. These three antibodies reacted with intestinal epithlial cell membranes but also cross-reacted with hepatocyte cytoplasm. The availability of intestinal specific monoclonal reagents will be of significant interest in studies of normal and abnormal intestinal physiology.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.19

年代:1988

数据来源: MAL

摘要:

A monoclonal antibody (MAb), designated H3, with specificity for hamster lymphocytes, was produced by somatic cell hybridization of myeloma Sp 2/0 and spleen cells of Balb/c mice immunized with suspensions of viable hamster thymocytes.The H3 MAb (IgG 3) reacted specifically with hamster thymocyte surface membranes (immunofluorescent assay). The antibody recognized a protein of an approximate molecular weight of 44,000 Daltons in immunoblots of hamster thymocyte extracts. The soluble H3 MAb presented potent mitogenic properties as indicated by the DNA synthesis induced inin vitrohamster lymphocyte cultures.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.33

年代:1988

数据来源: MAL

摘要:

The production and characterization of monoclonal antibodies (mAbs) against chicken B cell surface alloantigens, Bu-la and Bu-lb, is described. Flow cytometric analysis using these mAbs demonstrates that Bu-1 gene locus does not show allelic exclusion. Two color fluorescence analysis using simultaneous surface marker labeling and DNA staining shows that the expression of Bu-1 antigen is not restricted to a specific phase of the cell cycle. Earlier findings that Bu-l locus is not linked to the MHC locus are confirmed. Furthermore, data is presented that Bu-l antigen expression is not entirely B cell restricted. Apparently a fraction of larger mononuclear cells with typical light scatter characteristics in flow cytometry also bear this marker. MAbs against both allelic Bu-l antigens are beneficial tools e.g. in typing of chicken lines and in studies on chicken B cell differentiation.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.41

年代:1988

数据来源: MAL

摘要:

Two mouse IgM monoclonal antibodies, 177.1 and 179.3, are directed against glycophorin A, the major sialoglycoprotein of human erythrocytes. Both antibodies agglutinate blood group M and N erythrocytes equally well, both before and after treatment with neuraminidase or trypsin, but fail to agglutinate erythrocytes treated with papain. Antibody 179.3 agglutinates MiVII(K.T.) cells, whose glycophorin A probably contains some alterations in amino acid sequence between residues 46-56, but antibody 177.1 does not agglutinate these cells. Neither antibody agglutinates En(a-)G.W. cells, which lack glycophorin A completely. The hemagglutinating activity of antibody 177.1 is inhibited by purified glycophorin A and its chymotryptic glycopeptides CH1 (amino acid residues 1-64) and CH3 (amino acid residues 35-64), whereas the hemagglutinating activity of 179.3 is inhibited weakly by glycophorin A but not by chymotryptic peptides. These antibodies both are classified as anti-En(a-)FS but apparently bind different epitopes.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.49

年代:1988

数据来源: MAL

摘要:

Cross-linked Fibrin II was prepared using Kabi grade (L) fibrinogen. Fibrin plasmic digest was separated on Sepharose CL-6B. Fragments Mr 135-300 kDa were used to immunize 6-9 weeks old female BALB-c mice. A stable hybridoma secreting monoclonal antibody (MAb) TD-1 (IgG 2a, Kappa) was prepared by fusion using myeloma cells (P3-NSl/1-Ag4-1) and immunized cells. Fibrinogen and plasmin digest of fibrinogen in serial dilutions did not compete with the immunizing antigen. To prove that TD-1 binds specifically to cross-linked fibrin, immunoprecipitation with S. aureus and affinity chromatography were performed. In both experiments, we demonstrated that TD-1 binds specifically to a protein Mr>200 kDa which is found in XL-fibrin and not fibrinogen. Reduced samples showed the antibody bands (heavy and light chains) and three protein bands, Mr>80 kDa (γ-γ dimer), Mr>45 kDa (β chain of fragment D) and Mr>16 kDa (α chain from fragment D) were present. TD-1 reacted strongly with HPLC fraction of the immunizing antigen Mr 220 kDa (probably DD/E complex). Affinity binding constants (Scatchard Plot Analysis) were determined. The highest affinity was obtained with XL-fibrin fraction Mr 220 kDa, KD = 1.39 x 10-8and high molecular weight XL-fibrin fragments, KD = 1.6 x 10-7. Fragment DD had KD of 2.8 x 10-6. These results suggest that TD-1 is specific for the DD region of human cross-linked Fibrin

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.55

年代:1988

数据来源: MAL

摘要:

Different cell culture media were compared for their ability to support and promote the growth of stable hybridoma cell lines derived from three commonly used parental murine myelomas. Supplemented Dulbecco's modified Eagle's media (DMEM) and RPMI 1640 media were studied. The DMEM-based media were found to support greater numbers of cells for longer time periods than were the RPMI 1640-based media. Aminopterin supplemented medium was shown to be significantly less effective in supporting hybridoma reproduction and viability than medium without aminopterin. Antibody levels were directly related to cell concentration and viability regardless of the medium used for the hybridoma culture. An optimally formulated DMEM-based medium is suggested as the medium of choice for hybridoma propagation and maintenance.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.69

年代:1988

数据来源: MAL

摘要:

Hybridomas are as susceptible to mycoplasma contamination as animal cell cultures. Twoin vitromethods and onein vivopassage into mice were compared for efficiency of curing hybridomas from mycoplasma contaminations.Four contaminated clones were treated with the combined action of 5-bromouracil (5-BrUra) and the Hoechst 33258 followed by photosensitization. The otherin vitromethod involved the use of BM-cycline. The success of overcoming the mycoplasma contamination was dependent on the level of the initial contamination of the individual hybridoma. BM-cycline was more efficient than the photosensitization method. For the most contaminated hybridomas, 10 successive treatments with 5-BrUra were necessary as compared to six treatments with BM-cycline. Moreover, the use of BM-cycline reduced the contamination by as much as 50% after the first treatment. After twenty passages following the curing of hybridomas with BM-cycline, cells were stable and retained their specificity and secretion of their respective immunoglobulins. Whereas, treatment with 5-bromouracil, recurrence of contamination was observed in one of the four hybridomas after 10 passages following treatment. Decontamination after one passage in peritoneal cavity in mice was not always sufficient since one of four hybridomas remained contaminated. BM-cycline appears to be the method of choice since it is more efficient, less time consuming, simpler and less expensive. Mycoplasma strains that could be identified were of bovine origin: Mycoplasma arginini and Acholeplasma laidlawii.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.79

年代:1988

数据来源: MAL

摘要:

Monoclonal antibodies (MAbs) are useful reagents for the study of the structure and evolution of specific epitopes. Two MAbs of IgG1isotype, Tf-1 and Tf-2, which bind human transferrin have been produced and characterized. Both specifically recognize transferrin on immunoblots of serum. Proteolytic digestion with papain or chymotrypsin destroys the epitope recognized by Tf-1 but not Tf-2, demonstrating that the MAbs recognize distinct epitopes. Both epitopes are not recognized after treatment with 2-mercaptoethanol, suggesting that disulfide bond dependent tertiary structure is necessary for epitope integrity. Removal of carbohydrate moieties by treatment with trifluoromethane sulfonic acid likewise results in loss of reactivity. Neither MAb reacts with transferrin of mouse, rabbit or bovine origin. Both were tested for reactivity to a total of ten primate transferrins and showed different patterns of reaction. Tf-2 recognized human, chimpanzee and gorilla transferrins, whereas Tf-1 reacted with all Old World monkeys and one of three New World monkeys tested. Thus, Tf-1 and Tf-2 recognize transferrin epitopes with differential phylogenetic conservation and which are dependent not only on primary aminoacid sequence, but also upon tertiary structure and glycosylation.

ISSN:0272-457X    

DOI:10.1089/hyb.1988.7.87

年代:1988

数据来源: MAL