Antigen related to cell proliferation in malignant gliomas recognized by a human monoclonal antibody

Restricted access

✓ A human monoclonal antibody (CLN-IgG) was produced from a human-human hybridoma derived from lymphocytes of a patient with cervical carcinoma. The reactivities of this antibody with various human glioma tissues and cultured glioma cells and the characterization of the antigen recognized by CLN-IgG on malignant glioma cells were analyzed and reported. CLN-IgG reacted with various human glioma cells and glioma tissues, especially glioblastoma, but did not react with normal brain tissues or fetal brain tissues. A large amount of antigen recognized by CLN-IgG was expressed on cell membranes of undifferentiated glioma cells and of glioma cells at the G2/M tumor growth phase in cycling cells. Antigen recognized by CLN-IgG was detected in only one of seven samples of cyst fluid, and was not detected in 27 serum samples or 18 samples of cerebrospinal fluid from glioma patients. CLN-IgG exhibited antibody-dependent cell cytotoxicity against U-251MG glioma cells and primary cultured cells of glioblastomas and anaplastic astrocytomas. These data suggest that the antigen recognized by CLN-IgG might be related to cell proliferation in malignant gliomas. Thus, CLN-IgG might be useful for immunotherapy or immunoimaging of malignant gliomas.

Article Information

Address reprint requests to: Takashi Kokunai, M.D., Department of Neurosurgery, Kobe University School of Medicine, 7-5-1, Kusunoki-cho, Chuo-ku, Kobe 650, Japan.

© AANS, except where prohibited by US copyright law.

Headings

Figures

  • View in gallery

    Immunoperoxidase staining of glioblastoma (A), anaplastic astrocytoma (B), adult normal brain (C), and fetal brain (D) tissues with CLN-IgG. Nuclei were counterstained with hematoxylin. In A, a blood vessel is identified with an arrow. × 150.

  • View in gallery

    Immunofluorescence staining of fixed U-251MG cells with CLN-IgG. The reactive fluorescein isothiocyanate products with CLN-IgG, especially ruffles, were found on the cell membrane. Nuclei stained with propidium iodine. × 280.

  • View in gallery

    Two-color analysis of the vertical fluorescein isothiocyanate (FITC)-fluorescence (CLN-IgG) and the perpendicular propidium iodine-fluorescence (DNA) on KNS-42 (upper) and U-251MG (lower) cells by flow cytometry.

  • View in gallery

    Binding pattern of CLN-IgG on the antigen of U-251MG cells obtained with flow cytometric analysis. Upper Left: Exponentially growing cells. Lower Left: Cells treated with cyclic adenosine monophosphate (AMP). Upper Right: Cells synchronously accumulated at the G0/G1 phase. Lower Right: Cells collected at the G2/M phase by mitotic shaking. FITC = fluorescein isothiocyanate.

  • View in gallery

    Two-color analysis (left) and binding patterns (right) by flow cytometry of CLN-IgG on the antigen of primary cultured glioblastoma cells. FITC = fluorescein isothiocyanate; AMP = adenosine monophosphate.

  • View in gallery

    Immunoblotting analysis of the antigen in U-251MG cell lysate (lane 1), supernatant of U-251MG cells (lane 2), cyst fluid from glioblastomas (lanes 3 and 4), sera from glioma patients (lanes 5 and 6), and cerebrospinal fluid from glioma patients (lanes 7 and 8).

  • View in gallery

    Antibody-dependent cell cytotoxicity of CLN-IgG against U-251MG cells treated without (left) or with (right) cyclic adenosine monophosphate (AMP).

References

  • 1.

    Andreasen RBOlsson L: Antibody-producing human-human hybridomas. III. Derivation and characterization of two antibodies with specificity for human myeloid cells. J Immunol 137:108310901986Andreasen RB Olsson L: Antibody-producing human-human hybridomas. III. Derivation and characterization of two antibodies with specificity for human myeloid cells. J Immunol 137:1083–1090 1986

  • 2.

    Aotsuka YHagiwara H: Identification of a malignant cell associated antigen recognized by a human monoclonal antibody. Eur J Cancer Clin Oncol 24:8298381988Aotsuka Y Hagiwara H: Identification of a malignant cell associated antigen recognized by a human monoclonal antibody. Eur J Cancer Clin Oncol 24:829–838 1988

  • 3.

    Brown JPHewick RMHellström Iet al: Human melanoma-associated antigen p97 is structurally and functionally related to transferrin. Nature 296:1711731982Brown JP Hewick RM Hellström I et al: Human melanoma-associated antigen p97 is structurally and functionally related to transferrin. Nature 296:171–173 1982

  • 4.

    Carrasquillo JAKrohn KABeaumier Pet al: Prognosis of and therapy for solid tumors with radiolabeled antibodies and immune fragments. Cancer Treat Rep 68:3173281984Carrasquillo JA Krohn KA Beaumier P et al: Prognosis of and therapy for solid tumors with radiolabeled antibodies and immune fragments. Cancer Treat Rep 68:317–328 1984

  • 5.

    Cote RJMorrissey DMHoughton ANet al: Generation of human monoclonal antibodies reactive with cellular antigens. Proc Natl Acad Sci USA 80:202620301983Cote RJ Morrissey DM Houghton AN et al: Generation of human monoclonal antibodies reactive with cellular antigens. Proc Natl Acad Sci USA 80:2026–2030 1983

  • 6.

    Dillman ROShawler DLSobal REet al: Murine monoclonal antibody therapy in two patients with chronic lymphocytic leukemia. Blood 59:103610451982Dillman RO Shawler DL Sobal RE et al: Murine monoclonal antibody therapy in two patients with chronic lymphocytic leukemia. Blood 59:1036–1045 1982

  • 7.

    Edwards PAWSmith CMNeville AMet al: A human-human hybridoma system based on a fast-growing mutant of the ARH-77 plasma cell leukemia-derived line. Eur J Immunol 12:6416481982Edwards PAW Smith CM Neville AM et al: A human-human hybridoma system based on a fast-growing mutant of the ARH-77 plasma cell leukemia-derived line. Eur J Immunol 12:641–648 1982

  • 8.

    Glassy MCHandley HHHagiwara Het al: UC 729-6, a human lymphoblastoid B-cell line useful for generating antibody-secreting human-human hybridomas. Proc Natl Acad Sci USA 80:632763311983Glassy MC Handley HH Hagiwara H et al: UC 729-6 a human lymphoblastoid B-cell line useful for generating antibody-secreting human-human hybridomas. Proc Natl Acad Sci USA 80:6327–6331 1983

  • 9.

    Hagiwara HAotsuka Y: Cytotoxicity of mixed human monoclonal antibodies reacting to tumor antigens. Acta Paediatr Jpn 29:5525561987Hagiwara H Aotsuka Y: Cytotoxicity of mixed human monoclonal antibodies reacting to tumor antigens. Acta Paediatr Jpn 29:552–556 1987

  • 10.

    Hagiwara HSato GH: Human × human hybridoma producing monoclonal antibody against autologous cervical carcinoma. Mol Biol Med 1:2452521983Hagiwara H Sato GH: Human × human hybridoma producing monoclonal antibody against autologous cervical carcinoma. Mol Biol Med 1:245–252 1983

  • 11.

    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

  • 12.

    Köhler GMilstein C: Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:4954971975 (Letter)Köhler G Milstein C: Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497 1975 (Letter)

  • 13.

    Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:6806851970Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 1970

  • 14.

    Miller RAMaloney DGMckillop Jet al: In vivo effects of murine hybridoma monoclonal antibody in a patient with T-cell leukemia. Blood 58:78861981Miller RA Maloney DG Mckillop J et al: In vivo effects of murine hybridoma monoclonal antibody in a patient with T-cell leukemia. Blood 58:78–86 1981

  • 15.

    Miller RAMaloney DGWarnke Ret al: Treatment of B-cell lymphoma with monoclonal anti-idiotype antibody. N Engl J Med 306:5175221982Miller RA Maloney DG Warnke R et al: Treatment of B-cell lymphoma with monoclonal anti-idiotype antibody. N Engl J Med 306:517–522 1982

  • 16.

    Mitchell KFFuhrer JPSteplewski Zet al: Biochemical characterization of human melanoma cell surfaces: dissection with monoclonal antibodies. Proc Natl Acad Sci USA 77:728772911980Mitchell KF Fuhrer JP Steplewski Z et al: Biochemical characterization of human melanoma cell surfaces: dissection with monoclonal antibodies. Proc Natl Acad Sci USA 77:7287–7291 1980

  • 17.

    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

  • 18.

    Prasad KNHsie AW: Morphological differentiation of mouse neuroblastoma cells induced in vitro by dibutyryl adenosine 3′,5′-cyclic monophosphate. Nature 233 (New Biol): 1411421971Prasad KN Hsie AW: Morphological differentiation of mouse neuroblastoma cells induced in vitro by dibutyryl adenosine 3′5′-cyclic monophosphate. Nature 233 (New Biol):141–142 1971

  • 19.

    Ritz JPesando JMSallan SEet al: Serotherapy of acute lymphoblastic leukemia with monoclonal antibody. Blood 58:1411521981Ritz J Pesando JM Sallan SE et al: Serotherapy of acute lymphoblastic leukemia with monoclonal antibody. Blood 58:141–152 1981

  • 20.

    Robbins EMarcus PI: Mitotically synchronized mammalian cells: a simple method for obtaining large populations. Science 144:115211531964Robbins E Marcus PI: Mitotically synchronized mammalian cells: a simple method for obtaining large populations. Science 144:1152–1153 1964

  • 21.

    Saito HUchiyama KNakamura Iet al: Characterization of a human monoclonal antibody with broad reactivity to malignant tumor cells. JNCI 80:7287341988Saito H Uchiyama K Nakamura I et al: Characterization of a human monoclonal antibody with broad reactivity to malignant tumor cells. JNCI 80:728–734 1988

  • 22.

    Schlom JWunderlich DTeramoto YA: Generation of human monoclonal antibodies reactive with human mammary carcinoma cells. Proc Natl Acad Sci USA 77:684168451980Schlom J Wunderlich D Teramoto YA: Generation of human monoclonal antibodies reactive with human mammary carcinoma cells. Proc Natl Acad Sci USA 77:6841–6845 1980

  • 23.

    Sikora KAlderson TEllis Jet al: Human hybridomas from patients with malignant disease. Br J Cancer 47:1351451983Sikora K Alderson T Ellis J et al: Human hybridomas from patients with malignant disease. Br J Cancer 47:135–145 1983

  • 24.

    Sikora KWright R: Human monoclonal antibodies to lung-cancer antigens. Br J Cancer 43:6967001981Sikora K Wright R: Human monoclonal antibodies to lung-cancer antigens. Br J Cancer 43:696–700 1981

  • 25.

    Soule HRLinder EEdgington TS: Membrane 126-kilodalton phosphoglycoprotein associated with human carcinomas identified by a hybridoma antibody to mammary carcinoma cells. Proc Natl Acad Sci USA 80:133213361983Soule HR Linder E Edgington TS: Membrane 126-kilodalton phosphoglycoprotein associated with human carcinomas identified by a hybridoma antibody to mammary carcinoma cells. Proc Natl Acad Sci USA 80:1332–1336 1983

  • 26.

    Towbin HStaehelin TGordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:435043541979Towbin H Staehelin T Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354 1979

  • 27.

    Usuba OFujii YMiyoshi Iet al: Establishment of a human monoclonal antibody to Hanganutziu-Deicher antigen as a tumor-associated carbohydrate antigen. Jpn J Cancer Res 79:134013481988Usuba O Fujii Y Miyoshi I et al: Establishment of a human monoclonal antibody to Hanganutziu-Deicher antigen as a tumor-associated carbohydrate antigen. Jpn J Cancer Res 79:1340–1348 1988

  • 28.

    Warenius HMTaylor JWDurack BEet al: The production of human hybridomas from patients with malignant melanoma. The effect of pre-stimulation of lymphocytes with pokeweed mitogen. Eur J Cancer Clin Oncol 19:3473551983Warenius HM Taylor JW Durack BE et al: The production of human hybridomas from patients with malignant melanoma. The effect of pre-stimulation of lymphocytes with pokeweed mitogen. Eur J Cancer Clin Oncol 19:347–355 1983

TrendMD

Metrics

Metrics

All Time Past Year Past 30 Days
Abstract Views 35 35 3
Full Text Views 214 214 0
PDF Downloads 50 50 0
EPUB Downloads 0 0 0

PubMed

Google Scholar