Increased labeling of human melanoma cells in vitro using combinations of monoclonal antibodies recognizing separate cell surface antigenic determinants (original) (raw)
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Cancer research, 1985
Mouse monoclonal antibodies (mAb) detecting 13 distinct systems of surface antigens on cultured melanocytes and melanomas were tested for reactivity with panels of (a) normal and malignant cultured cells; (b) normal adult and fetal tissues; and (c) specimens of metastatic melanoma and other tumor types. The objectives of this study were to compare antigen expression in cultured versus noncultured cells, to develop a panel of mAbs that identify subsets of melanomas, and to provide requisite information about antibody specificity in preparation for the use of antibodies in the diagnosis, imaging, and therapy of melanoma. Five of the melanoma surface antigens have been well characterized biochemically [GD3, chondroitin sulfate proteoglycan, HLA Class II antigens, glycoprotein of molecular weight 130,000 (gp130), and glycoprotein Mr 95,000/protein Mr 97,000 (gp95/p97)]. Three antigens have been related to melanocyte differentiation (HLA Class II, M111/M231, and M144), and six provide ad...
Cell surface antigens of human melanoma identified by monoclonal antibody
Proceedings of the National Academy of Sciences, 1979
Mouse NS-1 myeloma cells were fused with spleen cells from mice that had been immunized with cells from a human melanoma, M1804. Hybrid cells were grown in selective medium and tested for production of antibody to surface antigens of M1804 cells. Three hybrids that produced antibodies that bound to the melanoma cells but not to autologous skin fibroblasts were cloned. Antibodies produced by two of the clones were cytotoxic to M1804 cells in the presence of rabbit complement. Extensive specificity tests showed that the antibodies produced by the clones bound strongly only to M1804 cells; significant, although weaker, binding occurred with 2 of 11 allogeneic melanomas. Apart from weak binding of the antibody produced by one of the clones to a breast carcinoma, binding assays of five carcinomas, one sarcoma, and fibroblasts from 17 individuals were negative, as were cytotoxic tests of 10
Analysis of two human monoclonal antibodies against melanoma
International Journal of Cancer, 1992
B cells derived from peripheral-blood lymphocytes (PBL) and tumor-infiltrating lymphocytes (TlL) from a patient with a high serum antibody titer to autologous melanoma were transformed with Epstein-Barr virus (EBV) and evaluated for reactivity against autologous tumor. B cells producing antibody reactive with autologous tumor and unreactive with normal fibroblasts were detected both in TIL and in PBL. One cell line derived from PBL and another derived from TIL sustained production of tumor-reactive antibody for I 0 weeks and over I 5 months respectively. The cell line derived from PBL, 2DI I, produced an antibody reactive with a tryprin-resistant antigen expressed on the cell membrane of autologous and allogeneic melanoma cell lines. The cell line derived from TIL, IF6, produced an antibody reactive with a cell-surface glycoprotein expressed by 5 autologous melanoma cell lines derived from 5 different metastases and I6/ I 9 allogeneic melanoma cell lines. IF6 also showed reactivity with cell lines derived from a blue nevus, a congenital nevus, an astrocytoma, and I /4 renal-cell carcinomas; but it was not reactive with 5 foreskin melanocyte cell lines, 2 normal fibroblast lines, 5 leukemia/lymphoma lines, 8 lung-cancer lines, 8 glioblastoma lines, or lines derived from I ovarian carcinoma, I colon carcinoma, I vulvar carcinoma, I fibrosarcoma, I murine melanoma, or 4 murine leukemia/ lymphomas. We describe here an antibody that detects a new melanoma specificity obtained by EBV transformation of tumorinfiltrating B cells.
Cancer Research, 1987
To identify melanoma associated antigens (MAAs) shared by human and guinea pig melanoma cells, a battery of murine monoclonal antibodies (MoAl>s) to human MAA and an antiserum to S100 protein were tested with four newly established guinea pig melanoma cell lines. Only the monoclonal antibodies 149.53 and 225.28 which recognize distinct deter minants of the human high molecular weight-MAA (HMW-MAA) re acted with all four guinea pig melanoma cell lines. To compare the binding site of MoAbs 149.53 and 225.28 with guinea pig and human melanoma cells, inhibition binding experiments were performed with antiidiotypic monoclonal antibodies which completely inhibit the binding of MoAbs 149.53 and 225.28 to human melanoma cells. The binding of M«Ah149.53 to guinea pig melanoma cells was partially inhibited by antiidiotypic MoAbs MF9-10 and MK1-180 which recognize distinct private idiotopes within the antigen combining site of MoAb 149.53. On the other hand the binding of MoAb 225.28 to guinea pig melanoma cells was completely inhibited by antiidiotypic MoAbs MF11-30 and I M-12 which recognize distinct private idiotopes within the antigen combining site of MoAb 225.28. These results suggest that the determinant recog nized by MoAb 149.53 on guinea pig melanoma cells is similar but not identical to that recognized on human melanoma cells, while the deter minants recognized by MoAb 225.28 on the two types of cells do not display any detectable differences under the experimental conditions tested. The target structure on the guinea pig melanoma cells identified by MoAbs 149.53 and 225.28 is a M, 280,000 molecule which has the same apparent molecular weight as one of the two subunits of the HMW-MAA synthesized by human melanoma cells. Sequential immunoprecipitation experiments with guinea pig melanoma cells showed that the determinant recognized by MoAb 149.53 is expressed on a subpopulation of the molecules recognized by MoAb 225.28. Immunohistochemical staining with MoAb 225.28 of a variety of different tissues from normal adult guinea pigs showed that the corresponding antigenic determinant is detectable only in basal cells of epidermis and hair follicles of skin. SIIMI protein, which is a cytoplasmic constituent of normal human melanocytes, benign nevi, and malignant melanocytes, was also detected in the cyto plasm of the four cultured guinea pig melanoma cells lines. The results of the present investigation may lead to a better understanding of the phylogenetic evolution of the human HMW-MAA and suggest that guinea pig melanoma may serve as a useful animal model for immunobiological studies and carcinogen-induced tumorigenesis investigations.
Criteria for selecting monoclonal antibodies with respect to accumulation in melanoma tissue
Cancer Immunology Immunotherapy, 1987
Immunohistology provides a necessary but insufficient criterion for selecting monoclonal antibodies (MAbs) capable of tumour targeting; in vivo. Additional selection procedures have been evaluated using a panel of anti-melanoma MAbs, including immunoreactivity of (labelled) MAbs, antibody affinity, kinetics of binding and release, apparent antigen density and accumulation in nude mouse transplants. According to these criteria, MAbs M.2.7.6 and M.2.9.4 showed the most favourable properties, i.e. high immunoreactivity and pronounced internalization into melanoma ceils. With MAbs M.2.10.15 and KG 6-56, moderate immunoreactivity and a binding pattern characterized by temperature dependence in the absence of internalization was observed. According to the paired label assay, all four MAbs showed specific accumulation into solid melanoma tissue. However, application in the patient still requires evaluation of the side effects of antigen cross-expression on normal human tissues.
Common human melanoma-associated antigen(s) detected by monoclonal antibodies
Cancer research, 1980
Hybridoma cells have been derived from a fusion between mouse myeloma cells (P3-NSI/1Ag4) and spleen cells from a mouse immunized with membrane-enriched fractions from the human melanoma cell line Me-43. Of the 26 hybrids obtained, seven secreted antibodies which reacted with the melanoma cell line used for immunoassay. The specificity of the antibodies produced by the seven positive hybrids was further investigated on 16 melanoma cell lines, 15 other tumors, and 14 lymphoblastoid cell lines. The antibodies from four positive hybrids showed a broad reactivity, whereas those from three hybrids reacted exclusively with melanoma cells. The antibodies from two of these three hybrids, alpha-Mel/5 and alpha-Mel/14, seem to be directed against common melanoma antigen(s) since they reacted with all (with one exception) of the 16 melanoma cell lines tested only with five of the 16 melanoma lines. Reciprocal binding inhibition tests using [3H]leeucine-labeled antibodies showed that alpha-Mel/...
Cell–cell adherence as a selection method for the generation of anti-melanoma monoclonal antibodies
Journal of Immunological Methods, 1997
The aim of the present study was to obtain monoclonal antibodies mAbs recognising human melanoma-associated antigens after immunisation of BALBrc mice with a 70-150 kDa membrane fraction from melanoma tumour tissues. Screening of specific antibody-producing hybridomas was performed using a novel cell-cell adherence method with the melanoma cell line M-14. Three mAbs of IgG1 isotype were selected: Mel-1, Mel-2 and Mel-3 which recognised the immunogen by ELISA and stained several melanoma cell lines positive in immunofluorescence. The molecular weight of the antigen was studied by different methods; a 170-kDa band was identified following immunoblotting of tumour lysate and a 72-kDa band was observed following immunoaffinity purification. Cell-cell adherence appears to be a reliable procedure for the generation of mAbs against native cellular antigens.