Human antiglioma monoclonal antibodies from patients with astrocytic tumors

Restricted access

✓ The current management of malignant gliomas is unsatisfactory compared to that of other solid tumors; the expected median survival period is less than 1 year with the patient undergoing conventional surgery, radiotherapy, and chemotherapy treatment. Immunological reagents could be a useful adjunct. Human monoclonal antibodies derived from patients with astrocytic tumors might recognize subtle antigenic specificities that would differ from those recognized by xenogeneic (murine) systems. Five hybridomas, designated as BT27/1A2, BT27/2A3, BT32/A6, BT34/A5, and BT54/B8, were produced from the fusion of peripheral blood lymphocytes of four patients with astrocytic tumors to the human myeloma-like cell line TM-H2-SP2. This cell line has a 46, XX karotype and is negative for hypoxanthine guanine phosphoribosyltransferase. All five human monoclonal antibodies produced 2.4 to 44 µg/ml of immunoglobulin M, had a similar but not identical pattern of reactivity against a panel of human tumor cell lines, and failed to react with normal human astrocytes. Labeling of four neuroectodermal tumor explant cultures by BT27/2A3 was demonstrated by flow cytometry. Karyotyping of three of the five hybridomas demonstrated that two were pseudodiploid (2–3n) and one hypodiploid (< 2n). The monoclonality of the hybridomas was evaluated by Southern blot analysis of JH gene rearrangements, revealing two types of rearrangements for each hybridoma, both consistent with monoclonality. Preliminary antigen characterization indicated that at least four of the five human monoclonal antibodies were directed to cell-surface glycolipids.

Article Information

Contributor Notes

Address reprint requests to: Gerald B. Price, Ph.D., McGill Cancer Centre, 3655 Drummond Street, Montreal, Quebec H3G 1Y6, Canada.

© AANS, except where prohibited by US copyright law.

Headings
References
  • 1.

    Barba DSaris SCHolder Cet al: Intratumoral LAK cell and interleukin-2 therapy of human gliomas. J Neurosurg 70:1751821989Barba D Saris SC Holder C et al: Intratumoral LAK cell and interleukin-2 therapy of human gliomas. J Neurosurg 70:175–182 1989

    • Search Google Scholar
    • Export Citation
  • 2.

    Bhondeley MKMehra RDMehra NKet al: Imbalances in T cell subpopulations in human gliomas. J Neurosurg 68:5895931988Bhondeley MK Mehra RD Mehra NK et al: Imbalances in T cell subpopulations in human gliomas. J Neurosurg 68:589–593 1988

    • Search Google Scholar
    • Export Citation
  • 3.

    Böyum A: Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest 21 (Suppl 97):77891968Böyum A: Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest 21 (Suppl 97):77–89 1968

    • Search Google Scholar
    • Export Citation
  • 4.

    Cahan LDIrie RFSingh Ret al: Identification of a human neuroectodermal tumor antigen (OFA-I-2) as ganglioside GD2. Proc Natl Acad Sci USA 79:762976331982Cahan LD Irie RF Singh R et al: Identification of a human neuroectodermal tumor antigen (OFA-I-2) as ganglioside GD2. Proc Natl Acad Sci USA 79:7629–7633 1982

    • Search Google Scholar
    • Export Citation
  • 5.

    Coleclough CPerry RPKarjalainen Ket al: Aberrant rearrangements contribute significantly to the allelic exclusion of immunoglobulin gene expression. Nature 290:3723781981Coleclough C Perry RP Karjalainen K et al: Aberrant rearrangements contribute significantly to the allelic exclusion of immunoglobulin gene expression. Nature 290:372–378 1981

    • Search Google Scholar
    • Export Citation
  • 6.

    Cote RJMorrissey DMHoughton ANet al: Specificity analysis of human monoclonal antibodies reactive with cell surface and intracellular antigens. Proc Natl Acad Sci USA 83:295929631986Cote RJ Morrissey DM Houghton AN et al: Specificity analysis of human monoclonal antibodies reactive with cell surface and intracellular antigens. Proc Natl Acad Sci USA 83:2959–2963 1986

    • Search Google Scholar
    • Export Citation
  • 7.

    Croce CMLinnenbach AHall Wet al: Production of human hybridomas secreting antibodies to measles virus. Nature 288:4884891980Croce CM Linnenbach A Hall W et al: Production of human hybridomas secreting antibodies to measles virus. Nature 288:488–489 1980

    • Search Google Scholar
    • Export Citation
  • 8.

    Douillard JYHoffman T: Enzyme-linked immunosorbent assay for screening monoclonal antibody production using enzyme-labeled second antibody. Methods Enzymol 92:1681741983Douillard JY Hoffman T: Enzyme-linked immunosorbent assay for screening monoclonal antibody production using enzyme-labeled second antibody. Methods Enzymol 92:168–174 1983

    • Search Google Scholar
    • Export Citation
  • 9.

    Ford AMMolgaard HVGreaves MFet al: Immunoglobulin gene organisation and expression in haemopoietic stem cell leukaemia. EMBO J 2:99710011983Ford AM Molgaard HV Greaves MF et al: Immunoglobulin gene organisation and expression in haemopoietic stem cell leukaemia. EMBO J 2:997–1001 1983

    • Search Google Scholar
    • Export Citation
  • 10.

    Gorer PAO'Gorman P: The cytotoxic activity of isoantibodies in mice. Transplant Bull 3:1421431956Gorer PA O'Gorman P: The cytotoxic activity of isoantibodies in mice. Transplant Bull 3:142–143 1956

    • Search Google Scholar
    • Export Citation
  • 11.

    Houghton ANBrooks HCote RJet al: Detection of cell surface and intracellular antigens by human monoclonal antibodies. Hybrid cell lines derived from lymphocytes of patients with malignant melanoma. J Exp Med 158:53651983Houghton AN Brooks H Cote RJ et al: Detection of cell surface and intracellular antigens by human monoclonal antibodies. Hybrid cell lines derived from lymphocytes of patients with malignant melanoma. J Exp Med 158:53–65 1983

    • Search Google Scholar
    • Export Citation
  • 12.

    Huang CCMoore GE: Chromosomes of 14 hemapoietic cell lines derived from peripheral blood of persons with and without chromosome anomalies. JNCI 43:111911281969Huang CC Moore GE: Chromosomes of 14 hemapoietic cell lines derived from peripheral blood of persons with and without chromosome anomalies. JNCI 43:1119–1128 1969

    • Search Google Scholar
    • Export Citation
  • 13.

    Irie RFSze LLSaxton RE: Human antibody to OFA-I, a tumor antigen, produced in vitro by Epstein-Barr virus-transformed human B-lymphoid cell lines. Proc Natl Acad Sci USA 79:566656701982Irie RF Sze LL Saxton RE: Human antibody to OFA-I a tumor antigen produced in vitro by Epstein-Barr virus-transformed human B-lymphoid cell lines. Proc Natl Acad Sci USA 79:5666–5670 1982

    • Search Google Scholar
    • Export Citation
  • 14.

    Katano MSidell NIrie RF: Human monoclonal antibody to a neuroectodermal tumor antigen (OFA-I-2). Ann NY Acad Sci 417:4274341983Katano M Sidell N Irie RF: Human monoclonal antibody to a neuroectodermal tumor antigen (OFA-I-2). Ann NY Acad Sci 417:427–434 1983

    • Search Google Scholar
    • Export Citation
  • 15.

    Kaufmann GZannis-Hadjopoulos MMartin RG: Cloning of nascent monkey DNA synthesized early in the cell cycle. Mol Cell Biol 5:7217271985Kaufmann G Zannis-Hadjopoulos M Martin RG: Cloning of nascent monkey DNA synthesized early in the cell cycle. Mol Cell Biol 5:721–727 1985

    • Search Google Scholar
    • Export Citation
  • 16.

    Korsmeyer SJArnold ABakhshi Aet al: Immunoglobulin gene rearrangement and cell surface antigen expression in acute lymphocytic leukemias of T cell and B cell precursor origins. J Clin Invest 71:3013131983Korsmeyer SJ Arnold A Bakhshi A et al: Immunoglobulin gene rearrangement and cell surface antigen expression in acute lymphocytic leukemias of T cell and B cell precursor origins. J Clin Invest 71:301–313 1983

    • Search Google Scholar
    • Export Citation
  • 17.

    Littlefield JW: Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145:7097101964Littlefield JW: Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145:709–710 1964

    • Search Google Scholar
    • Export Citation
  • 18.

    Livingston PONatoli EJCalves MJet al: Vaccines containing purified GM2 ganglioside elicit GM2 antibodies in melanoma patients. Proc Natl Acad Sci USA 84:291129151987Livingston PO Natoli EJ Calves MJ et al: Vaccines containing purified GM2 ganglioside elicit GM2 antibodies in melanoma patients. Proc Natl Acad Sci USA 84:2911–2915 1987

    • Search Google Scholar
    • Export Citation
  • 19.

    Lowry OHRosebrough NJFarr ALet al: Protein measurement with the Folin phenol reagent. J Biol Chem 193:2652751951Lowry OH Rosebrough NJ Farr AL et al: Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275 1951

    • Search Google Scholar
    • Export Citation
  • 20.

    Mahaley MS JrBrooks WHRoszman TLet al: Immunobiology of primary intracranial tumors. Part 1. Studies of the cellular and humoral general immune competence of brain-tumor patients. J Neurosurg 46:4674761977Mahaley MS Jr Brooks WH Roszman TL et al: Immunobiology of primary intracranial tumors. Part 1. Studies of the cellular and humoral general immune competence of brain-tumor patients. J Neurosurg 46:467–476 1977

    • Search Google Scholar
    • Export Citation
  • 21.

    Mahaley MS JrSteinbok PAronin Pet al: Immunobiology of primary intracranial tumors. Part 4. Levamisole as an immune stimulant in patients and in the ASV glioma model. J Neurosurg 54:2202271981Mahaley MS Jr Steinbok P Aronin P et al: Immunobiology of primary intracranial tumors. Part 4. Levamisole as an immune stimulant in patients and in the ASV glioma model. J Neurosurg 54:220–227 1981

    • Search Google Scholar
    • Export Citation
  • 22.

    Maniatis TFritsch EFSambrook J: Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratories1982Maniatis T Fritsch EF Sambrook J: Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratories 1982

    • Search Google Scholar
    • Export Citation
  • 23.

    Max EE: Immunoglobulins: molecular genetics in Paul WE (ed): Fundamental Immunology. New York: Raven Press1984 pp 167204Max EE: Immunoglobulins: molecular genetics in Paul WE (ed): Fundamental Immunology. New York: Raven Press 1984 pp 167–204

    • Search Google Scholar
    • Export Citation
  • 24.

    Nottenburg CWeissman IL: Cµ gene rearrangement of mouse immunoglobulin genes in normal B cells occurs on both the expressed and nonexpressed chromosomes. Proc Natl Acad Sci USA 78:4844881981Nottenburg C Weissman IL: Cµ gene rearrangement of mouse immunoglobulin genes in normal B cells occurs on both the expressed and nonexpressed chromosomes. Proc Natl Acad Sci USA 78:484–488 1981

    • Search Google Scholar
    • Export Citation
  • 25.

    Olsson LKaplan HS: Human-human hybridomas producing monoclonal antibodies of predefined antigenic specificity. Proc Natl Acad Sci USA 77:542954311980Olsson L Kaplan HS: Human-human hybridomas producing monoclonal antibodies of predefined antigenic specificity. Proc Natl Acad Sci USA 77:5429–5431 1980

    • Search Google Scholar
    • Export Citation
  • 26.

    Phillips JAlderson TSikora Ket al: Localisation of malignant glioma by a radiolabelled human monoclonal antibody. J Neurol Neurosurg Psychiatry 46:3883921983Phillips J Alderson T Sikora K et al: Localisation of malignant glioma by a radiolabelled human monoclonal antibody. J Neurol Neurosurg Psychiatry 46:388–392 1983

    • Search Google Scholar
    • Export Citation
  • 27.

    Pontecorvo G: Production of mammalian somatic cell hybrids by means of polyethylene glycol treatment. Somat Cell Genet 1:3974001975Pontecorvo G: Production of mammalian somatic cell hybrids by means of polyethylene glycol treatment. Somat Cell Genet 1:397–400 1975

    • Search Google Scholar
    • Export Citation
  • 28.

    Ravetch JVSiebenlist UKorsmeyer Set al: Structure of the human immunoglobulin µ locus: characterization of embryonic and rearranged J and D genes. Cell 27:5835911981Ravetch JV Siebenlist U Korsmeyer S et al: Structure of the human immunoglobulin µ locus: characterization of embryonic and rearranged J and D genes. Cell 27:583–591 1981

    • Search Google Scholar
    • Export Citation
  • 29.

    Rudders RADhillon SKrontiris TG: Clonal analysis of untreated non-Hodgkin's lymphoma utilizing immunoglobulin gene rearrangement and immunophenotype. Cancer Res 48:627262771988Rudders RA Dhillon S Krontiris TG: Clonal analysis of untreated non-Hodgkin's lymphoma utilizing immunoglobulin gene rearrangement and immunophenotype. Cancer Res 48:6272–6277 1988

    • Search Google Scholar
    • Export Citation
  • 30.

    Salcman M: Survival in glioblastoma: historical perspective. Neurosurgery 7:4354391980Salcman M: Survival in glioblastoma: historical perspective. Neurosurgery 7:435–439 1980

    • Search Google Scholar
    • Export Citation
  • 31.

    Sikora KAlderson TPhillips Jet al: Human hybridomas from malignant gliomas. Lancet 1:11141982Sikora K Alderson T Phillips J et al: Human hybridomas from malignant gliomas. Lancet 1:11–14 1982

    • Search Google Scholar
    • Export Citation
  • 32.

    Stewart SSPrice GB: Realtime acquisition, storage, and display of correlated three-parameter flow cytometric data. Cytometry 7:82881986Stewart SS Price GB: Realtime acquisition storage and display of correlated three-parameter flow cytometric data. Cytometry 7:82–88 1986

    • Search Google Scholar
    • Export Citation
  • 33.

    Sullivan AKTeskey LMCampling Bet al: The potential of hybridomas to study autoantibodies in man in Russ LCarlton D (eds): Hybridoma Technology. Toronto: Ortho Pharmaceuticals Canada1982 pp 6368Sullivan AK Teskey LM Campling B et al: The potential of hybridomas to study autoantibodies in man in Russ L Carlton D (eds): Hybridoma Technology. Toronto: Ortho Pharmaceuticals Canada 1982 pp 63–68

    • Search Google Scholar
    • Export Citation
  • 34.

    Suter LBrüggen JSorg C: Use of an enzyme-linked immunosorbent assay (ELISA) for screening of hybridoma antibodies against cell surface antigens. J Immunol Methods 39:4074111980Suter L Brüggen J Sorg C: Use of an enzyme-linked immunosorbent assay (ELISA) for screening of hybridoma antibodies against cell surface antigens. J Immunol Methods 39:407–411 1980

    • Search Google Scholar
    • Export Citation
  • 35.

    Svennerholm LFredman P: A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617:971091980Svennerholm L Fredman P: A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617:97–109 1980

    • Search Google Scholar
    • Export Citation
  • 36.

    Tokumaru TCatalano LW Jr: Elevation of serum immunoglobulin M (IgM) level in patients with brain tumors. Surg Neurol 4:17211975Tokumaru T Catalano LW Jr: Elevation of serum immunoglobulin M (IgM) level in patients with brain tumors. Surg Neurol 4:17–21 1975

    • Search Google Scholar
    • Export Citation
TrendMD
Metrics

Metrics

All Time Past Year Past 30 Days
Abstract Views 64 64 1
Full Text Views 124 74 0
PDF Downloads 108 68 0
EPUB Downloads 0 0 0
PubMed
Google Scholar