Immunoreactivity of human MAb BT32/A6 with neuroepithelial tumors (original) (raw)
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Human glioma-associated antigens detected by monoclonal antibodies
Cancer research, 1981
Hybridoma cells were derived from a fusion between mouse P3x63/Ag8 myeloma cells and spleen cells from a mouse immunized with whole cells of a human malignant glioma line. Of 345 hybrids obtained, 36 secreted antibodies that reacted with the glioma cell line used for immunization as assayed by an indirect antibody-binding radioimmunoassay. After a first screening for the absence of reactivity on two nongliogenous cell lines, 3 hybrids were selected and cloned by limiting dilution. The specificity of these monoclonal antibodies was then investigated on a panel of 18 cell lines derived from human malignant gliomas, 18 cell lines from nongliogenous neoplasms, as well as normal peripheral blood lymphocytes, normal skin fibroblasts, and normal spermatozoa. The monoclonal antibodies from two positive hybrids, BF7 and GE2, reacted exclusively with glioma cells and appeared to be directed against common malignant glioma antigen(s). BF7 antibodies bound to 13 and GE2 to 17 of 18 glioma cell ...
Production and Characterization of Two Human Glioma Xenograft-localizing Monoclonal Antibodies
Cancer Research
Multiple fusions following immunization of athymic mice with the extensively characterized human glioma cell line D-54 MG resulted in the selection of several antibodies (Mabs) highly reactive with tumors of neuroectodermal origin and unreactive with normal nervous system tissue. Two Mabs, C12 and D12, which localized specifically to tumors in athymic mouse-human glioma xenograft paired label localization assays, are IgG3 antibodies; both bind readily to staphylococcal protein A in column purification and radioimmunoprecipitation procedures. Both iodinate via the chloramine-T method yielding I2*l-immunoreactive product by direct cell surface radioimmunoassay and absorption assay. By indirect cell surface radioimmunoassay, a cultured cell line panel consisting of 17 gliomas, 3 medulloblastomas, 2 neuroblastomas, 2 melanomas, and 2 fetal and 2 adult brain-derived cell lines was examined; the two Mabs were highly similar but distinct in their reactivity profiles. Each was positive with >47% of the gliomas tested (C12, 9 of 17; D12, 8 of 17); and with 1 of 3 medulloblastomas, 1 of 2 melanomas, and cell lines derived from 12-and 16-week-gestation human fetal brain. No reactivity was observed with neuroblastoma or adult brain-derived cell lines or with neutral glycolipids and gangliosides extracted from D-54 MG xenografts or human glioma cell lines. Notable extraneuroectodermal reactivity included that of Mab D12 for splenic trabeculae and the spermatids and Sertoli cells in the testes. Following immunoprecipitation of ('Hjleucine labeled cell membrane preparations, Mabs C12 and D12 have consis tently yielded unique bands in the M, 180,000 and M, 88,000 regions respectively. When used in paired label localization experiments in s.c. D-54 MG xenograft-bearing athymic mice, Mabs C12 and D12 demon strate similar localization patterns, attaining peak localization indices at day 3 (D12) or 4 (C12); the maximum percentage of injected Mab bound to tumor ranged from 57°(D12) to 8% (C12). The peak tumor/normal brain localization ratios (167-181) attained by these Mabs at days 1-2 followed by their rapid clearance suggest that these Mabs are potentially useful imaging and therapeutic agents for further investigation. . 3The abbreviations used are: Mab, monoclonal antibody; CS-R1A, cell surface radioimmunoassay; FCS. fetal calf serum; and ZO, zinc option medium supple mented with glutamine and 10-20% PCS as detailed in the text; ABC, avidinbiotin complex; LI, localization index. Immunization, Fusion, Cloning, and Immunoglobulin Preparation.
Immunolocalization of neuroblastoma using radiolabeled monoclonal antibody UJ13A
The Journal of Pediatrics, 1984
The monoclonal antibody UJI 3A, raised after immunization oJ" mice with human Jetal brain, recognized an antigen expressed on human neuroblastoma cell lines and fresh tumors. Antibody was purified and radiolabeled with iodine isotopes using chloramine-T. In preclinical studies, ~251-labeled UJI 3A was injected intravenously into nude mice bearing xenografts of human neuroblastoma. Radiolabeled UJI 3A uptake by the tumors was Jour to 23 times greater than that by blood. In control animals, injected with a similar quantity of a monoclonal antibody known not to bind to neuroblastoma cells in vitro , there was no selective tumor uptake. Nine patients with histologically confirmed neuroblastoma each received 100 to 300 gg UJI 3A radiolabeled with 1 to 2.8 mCi ~231 or J3~l. Sixteen positive sites were visible on gamma scans 1 to 7 days aider injection: 15 were primary or secondary tumor sites, and one was a false positive; there were two false negatives. In two of the 15 positive sites, tumor had not been demonstrated by other imaging techniques; these were later confirmed as areas of malignant infiltration. No toxicity was encountered. (J PEDIATR
Journal of Neuroimmunology, 1995
The reactivity of a mAb (M16) raised against a small cell lung carcinoma line is described. Ml6 identifies a surface antigen expressed on cells of neuroectodermal origin following activation, as well as neoplastic transformation. Ml6 antigen expression is increased on retinoblastoma and neuroblastoma cell lines upon 'in vitro' stimulation and it is induced 'in vivo' on glial cells activated following brain injury. Furthermore, glial tumors show levels of Ml6 molecule expression increasing with the degree of malignancy, and in a retinoblastoma cell line, the expression of Ml6 was inversely related to the level of HLA-Class I and N-CAM antigens. The Ml6 antigen may represent a marker of both activation and neoplastic progression for neuroectodermal cells.
Human Leukocyte Antigen and Antigen Processing Machinery Component Defects in Astrocytic Tumors
Clinical Cancer Research, 2005
To determine the frequency of abnormalities in human leukocyte antigen (HLA) and antigen processing machinery (APM) component expression in malignant brain tumors. This information may contribute to our understanding of the immune escape mechanisms used by malignant brain tumors because HLA antigens mediate interactions of tumor cells with the host's immune system. Eighty-eight surgically removed malignant astrocytic tumors, classified according to the WHO criteria, were stained in immunoperoxidase reactions with monoclonal antibody recognizing monomorphic, locus-specific, and allospecific determinants of HLA class I antigens, beta2-microglobulin, APM components (LMP2, LMP7, TAP1, TAP2, calnexin, calreticulin, and tapasin), and HLA class II antigens. HLA class I antigens were lost in approximately 50% of the 47 glioblastoma multiforme (GBM) lesions and in approximately 20% of the 18 grade 2 astrocytoma lesions stained. Selective HLA-A2 antigen loss was observed in approximately 80% of the 24 GBM lesions and in approximately 50% of the 12 grade 2 astrocytoma lesions stained. HLA class I antigen loss was significantly (P < 0.025) correlated with tumor grade. Among the APM components investigated, tapasin expression was down-regulated in approximately 20% of the GBM lesions analyzed; it was associated, although not significantly, with HLA class I antigen down-regulation and tumor grade. HLA class II antigen expression was detected in approximately 30% of the 44 lesions analyzed. The presence of HLA antigen defects in malignant brain tumors may provide an explanation for the relatively poor clinical response rates observed in the majority of the T cell-based immunotherapy clinical trials conducted to date in patients with malignant brain tumors.
Journal of Neuroimmunology, 1986
Summaff 69−c15isahighlyimmunogeniccelllinederivedfromanaviansarcomavirus(ASV)−inducedastrocytomainF−344rats.Monoclonalantibody(Mab)productionwasattemptedbyfusingF−344ratsplenocytesandmouseP3×63/Ag8.653myelomacellsafterasyngeneicimmunizationprotocol.336fusioncloneswerescreenedbycellsurfaceradioimmunoassay(CS−RIA)againsttheimmunizingline69-c15 is a highly immunogenic cell line derived from an avian sarcoma virus (ASV)-induced astrocytoma in F-344 rats. Monoclonal antibody (Mab) production was attempted by fusing F-344 rat splenocytes and mouse P3 × 63/Ag8.653 myeloma cells after a syngeneic immunization protocol. 336 fusion clones were screened by cell surface radioimmunoassay (CS-RIA) against the immunizing line 69−c15isahighlyimmunogeniccelllinederivedfromanaviansarcomavirus(ASV)−inducedastrocytomainF−344rats.Monoclonalantibody(Mab)productionwasattemptedbyfusingF−344ratsplenocytesandmouseP3×63/Ag8.653myelomacellsafterasyngeneicimmunizationprotocol.336fusioncloneswerescreenedbycellsurfaceradioimmunoassay(CS−RIA)againsttheimmunizingline69-c15, rat kidney fibroblast line 203−cllandWalkerratcarcinomaline.Mabs7G4,9F1,10E3and10E7whichreactedonlywith203-cll and Walker rat carcinoma line. Mabs 7G4, 9F1, 10E3 and 10E7 which reacted only with 203−cllandWalkerratcarcinomaline.Mabs7G4,9F1,10E3and10E7whichreactedonlywith69-c15 were chosen. Further analysis demonstrated that these Mabs reacted only with rat (13/23 astrocytomas, 2/4 gliomas, 1/11 neurinomas) or mouse (2/10 astrocytomas) neurogenic tumor cells induced by both viral and chemical agents. Reciprocal competition assays suggested that 7G4, 9F1 and 10E3 recognized the same epitope and that 10E7 reacted with a spatially close determinant. Antigen activity could not be found in adult rat tissues (brain, heart, lung, liver, kidney, spleen, thymus, intestine, muscle and peripheral nerve) and fetal brain (8, 12, 20 days gestation) by either absorption analysis or tissue staining. Preliminary characterization indicated that the epitope may be polypeptide-associated. Further antigen purification and tumor localization can be attempted with these Mabs.
Journal of Neuroimmunology, 1995
A monoclonal antibody 6DS, against a human glioblastoma multiforme cell line U-87MG recognizes a tumor-specific, cell surface antigen of human glioblastoma cell lines. Partial cross-reactivity is observed with two human neuroblastoma cell lines, SK-N-SH and SK-N-MC, with little or no reactivity towards a rat glioma cell line C6 or normal human adult and fetal brain tissues. The antibody recognizes an antigen of molecular mass 38 kDa as inferred from Western blot analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the immunoprecipitate. The monoclonal antibody 6DS, inhibits both the attachment to substratum and growth of U-87MG cells. It strongly cross-reacts with xenotransplants of U-87MG cells and inhibits tumorigenesis (subcutaneous implants of U-87MG cells) in nude mice.
Immunoglobulin classes G,A,M in brain tumours
JPMA. The Journal of the Pakistan Medical Association, 1992
Serum of 37 brain tumour patients was studied for, Total protein, Protein electrophoresis, IgG, IgA, and IgM in an attempt to ascertain humoral immune response. Findings were compared with 20 healthy subjects, matched for age, sex and socioeconomic status. There was significant rise in alpha-II globulin, while IgG was suppressed in tumour patients. Decrease of IgG was more marked in patients with malignant tumours. Immunoglobulin A was low in children when compared with adults. Immunoglobulin M remained unchanged (JPMA 42: 157, 1992).