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  • 1
    ISSN: 1432-0851
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A 3-dimensional tumor growth inhibition assay [18] has been adapted to test the cytotoxic activity of a panel of monoclonal antibodies directed to various antigenic determinants on the surface of mouse mammary tumor cells. Target cells can be prepared from either cultured cells or from pieces of fresh tumor. Antibody and complement are added when cells are growing actively and cell growth can be measured, non destructively, over a 7–10-day period. Effective diffusion of antibody through collagen gel and binding to target cells embedded in the gel is demonstrated by indirect immunofluorescent staining. The specificity of monoclonal antibody AMT 101 cytotoxicity for mouse mammary tumor cells is the same in trypan blue exclusion assays of single-cell suspensions as in collagen gel assays, with complete killing seen in the collagen gel assay only. The collagen gel assay allows the testing of repeated treatments in vitro, as well as combined treatment with multiple antibodies. It also allows cell-cell interaction and preserves all cell components in the tumor. The collagen gel assay has potential as a method of predicting the outcome of monoclonal antibody treatment of solid tumors.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0851
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary High-resolution two-dimensional polyacrylamide gel electrophoresis (PAGE) was employed to reveal tumor-associated polypeptide changes, using the BALB/c C4 line mouse mammary model system, for which phenotypic and immunogenic alterations accompanying tumor progression are well defined. In the first set of experiments, polypeptide patterns from 20 µg whole tissue lysates of normal mammary gland, C4 preneoplastic hyperplatic alveolar nodule outgrowth (HAN) and spontaneous tumor from C4 HAN were compared. In order to normalize for differential cellularity and extracellular protein content in the whole tissues, our analysis included polypeptide patterns from serum, increased concentration of protein from whole normal mammary gland, and primary cultures of epithelial cells from normal gland, HAN and tumor. Using a computer-based image-analysis system, 90 polypeptides were identified in C4 tumor that were absent in C4 HAN, normal mammary gland and serum. None of the 90 polypeptides could be shown to represent a definite qualitative change in the protein composition of tumor epithelium as they were found to be either present in a higher concentration of protein from whole normal gland, or present in the primary epithelial culture from HAN, or absent in the primary epithelial culture from tumor. Conversely in the second set of experiments, when epithelial cultures were used as the starting point for comparisons to locate tumor-associated polypeptides, none of the 15 polypeptides that were present in cultures from three different tumors, and absent in the culture from normal mammary gland was specific to C4 tumor, as they were present in whole tissues of normal gland. Thus our experimental approach detected significant quantitative but no qualitative polypeptide changes in whole tumor tissue, or in tumor-derived epithelial cell cultures. This finding may reflect the limitations of the two-dimensional PAGE method, and warrants caution in the use of such gel analysis alone to identify tumor-associated proteins.
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  • 3
    ISSN: 1432-0851
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Tumor infiltrating lymphocytes (TIL) were isolated by centrifugal elutriation from C4 mouse mammary tumors and characterized with regard to phenotype and natural killer (NK) activity. Tumors that had arisen spontaneously in prenoplastic hyperplastic alevolar nodules and tumors that had been passaged one to two times in either naive or presensitized mice were studied. Mice were sensitized by limited s.c. tumor growth and subsequent surgical removal of the tumor. The total numbers of T or B cells in the infiltrates were similar in spontaneous tumors and in passaged tumors from either naive or sensitized mice. The ratio of L3T4-positive to lyt-2-positive cells was reduced, however, from 1.10±0.2 in spontaneous tumors to 0.53±0.28 or 0.48±0.04 in passaged tumors from untreated or sensitized mice. The site of tumor implantation, whether intramammary fat pad or s.c., did not affect the profiles of the infiltrates. The TIL from both spontaneous and passaged tumors demonstrated enhanced NK activity relative to peripheral lymphoid cells. The TIL of passaged tumors sensitized mice, however, had lower NK activity than those from naive mice.
    Type of Medium: Electronic Resource
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