Anti-(glioma surface antigen) monoclonal antibody G-22 recognizes overexpressed CD44 in glioma cells

Okada, Hideho; Yoshida, Jun; Seo, Hisao; Wakabayashi, Takashi; Sugita, Kenichiro; Hagiwara, Masatoshi

doi:10.1007/bf01519984

Cited by 12 publications

(4 citation statements)

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“…Many monoclonal antibodies against gliomas, prepared by immunizing mice with glioma cells, have been reported. Some of those antibodies have been analysed with regard to their target molecules (Schold et al, 1993;Okada et al, 1994;Romejin et al, 1994). Mouse MAbs against gliomas have also been applied in experimental models.…”

Section: Discussionmentioning

confidence: 99%

Monoclonal antibody ONS-M21 recognizes integrin α3 in gliomas and medulloblastomas

et al. 1998

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Medulloblastomas are the most common type of primitive neuroectodermal tumours observed in the central nervous system of children. In spite of recent advances in therapy, including microsurgery, combined chemotherapy and radiotherapy, the longterm prognosis for patients with these tumours is not satisfactory (Evans et al, 1990), pointing to a need for more specific and efficient therapies. Antibodies that recognize medulloblastoma cells are expected to act as diagnostic and therapeutic reagents. Therefore, we attempted to produce a medulloblastoma-specific monoclonal antibody (MAb). Immunization of mice with ONS-76 medulloblastoma cells (Tamura et al, 1989) successfully produced the antibodies referred to as ONS-M21 MAb (Moriuchi et al, 1993). ONS-M21 MAb reacted with a surface antigen of most glioma and medulloblastoma cell lines, and it also reacted with surgical specimens of gliomas and medulloblastomas. On the other hand, this MAb did not show cross-reactivity with peripheral blood cells or normal brain tissue. However, mouse antibodies are highly immunogenic in humans, and, hence, their clinical use may be limited. We genetically engineered a recombinant, humanized version of ONS-M21 MAb (hONS-M21 Ab) by means of CDR grafting (Ohtomo et al, 1995) in order to eliminate immunogenicity. This humanized Ab competed with ONS-M21 MAb for binding to glioma cells. In addition, this Ab, when labelled with an isotope, was capable of depicting tumour location by an autoradiographic technique. Thus, humanized Ab possesses various advantages that make it attractive for clinical applications.As the target molecule of ONS-M21 remains unknown, identification of this antigen is a necessary step before any clinical use of this antibody can be attempted. In the present study, we purified ONS-M21 antigen from ONS-76 cell lysates by means of an Abbinding affinity column, analysed the amino acid sequences of the ONS-M21 binding protein and identified the target molecule of ONS-M21 MAb. In addition, the mRNA level of this antigen was analysed by reverse transcription-polymerase chain reaction (RT-PCR) in surgical specimens of gliomas and medulloblastomas. MATERIALS AND METHODS Cell culture conditions and preparation of the monoclonal antibodyThe human medulloblastoma cell line ONS-76 was established in our laboratory (Tamura et al, 1989; Institution for fermentation, Osaka, No. 50355), maintained in Dulbecco's modified Eagle medium (DMEM) with 10% heat-inactivated fetal bovine serum (FBS) and 50 µg ml -1 gentamycin, and incubated in a humidified 10% carbon dioxide-in-air atmosphere at 37°C. ONS-76 cells were collected using a cell scraper and washed with cold Trisbuffered saline (TBS; 10 mmol l -1 Tris-Cl, 150 mmol l -1 NaCl, pH 8.0). The cells were resuspended in a detergent solution (TBS with 0.5% Tween-20 and 1 mg ml -1 p-amidinophenyl methanesulphonyl fluoride hydrochloride) at a concentration of 10 6 cells ml -1 , and then shaken for 2 h at 4°C to elute the cell extract. After centrifuging (3000 r.p.m., 20 min), the supernata...

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Section: Discussionmentioning

confidence: 99%

Monoclonal antibody ONS-M21 recognizes integrin α3 in gliomas and medulloblastomas

et al. 1998

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“…The sequence of total 54 amino-acid-residues was identical to the portion 55-108 of human CD44. We concluded that the smallest spliced form (85 kDa) of CD44 is strongly expressed in glioma cells (Okada et al, 1994).…”

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confidence: 99%

Suppression of CD44 expression decreases migration and invasion of human glioma cells

et al. 1996

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A unique characteristic of malignant gliomas is their invasive behaviour into surrounding normal brain tissue, making glioma-development a life-threatening disease despite today's advanced multimodality treatment. We have been exploring a molecular targeting strategy against malignant gliomas. By immunising mice with a human glioma cell line SK-MG-4 we obtained a monoclonal antibody (MAb), G-22, that reacts specifically with most human glioma cell lines (Wakabayashi et af., 1988). The antibody has been utilised for various clinical applications, including target imaging (Yoshida et af., 1990) and cerebrospinal-fluid diagnosis of human gliomas (Wakabayashi et al., 1988). We have purified a 85-kDa protein as the antigen of G-22 by imrnuno-affinity chromatography. The sequence of total 54 amino-acid-residues was identical to the portion 55-108 of human CD44. We concluded that the smallest spliced form (85 kDa) of CD44 is strongly expressed in glioma cells (Okada et al., 1994).The CD44 (Okada et al., 1994). In addition, the isoform of CD44 expressed on glioma cells was confirmed to be the smallest spliced form (Li etal., 1993) with a higher affinity for hyaluronic acid than other isoforms. Indeed, the human glioblastoma cell line U-373 MG was found to exhibit binding to hyaluronic acid via CD44 (Asher and Bignami, 1992). This 85-kDa cell-surface glycoprotein has been suggested to be closely associated with actin filaments through its cytoplasmic domain, consistent with the role of CD44H in cell motility and migration (Lacy and Underhill, 1987). Generally the biological attributes leading to cell invasion include adhesion to tissue structures such as ECMs, motility to generate migration, and the ability to influence the local environment so as to create space into which to move (Liotta and Stetler-Stevenson, 1991) by the secretion of proteases (Rao et al., 1993). Considering that hyaluronic acid is a major ECM component in human brain and that astrocytes have been found to produce hyaluronic acid (Asher and Bignami, 1991), high expression of CD44H on glioma cells may be expected to play a critical role in their migration and invasive behaviour.In the present study, we established a CD44-anti-senseexpression cell line named U-251A1 and investigated the migratory response of human glioma cell lines to hyaluronicacid-coated substrates in relation to CD44 expression in vitro. In addition, U-251A1 and its sense-transfected control cell U-251S1 were injected into brains of nude mice to observe differences in growth and invasion patterns in vivo. Our findings suggest that both migratory and invasive properties of human glioma cells depend upon the level of expression of CD44H. MATERIAL AND METHODS Cell finesHuman glioma cell lines GL 9 and GL 11 were derived from glioma patients and established at the Tissue Culture Laboratories of Kamo Hospital (Toyota, Japan) (Wakabayashi et aL, 1988). All cell lines used here were maintained in Dulbecco's modified Eagle's medium (DMEM, GIBCO, Paisley, UK) supplemented with 10% FCS, st...

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“…Several studies have found CD44 to be linked with cell-cell adhesion, cell adhesion to ECM, cell migration, as well as cancer development and metastasis [12,[57][58][59]. Nevertheless, excessive quantities of CD44s secretion have been observed in primary human gliomas as well as human glioma cell lines [4,[60][61][62]. Studies have demonstrated that, CD44 is secreted by astrocytes, mesenchymal cells, epithelial cells, as well as Schwann cells [63][64][65].…”

Section: Hyaluronic Acid Receptors and Gliomamentioning

confidence: 99%

The Pivotal Novel Pathogenic Roles of Hyaluronic Acid and its Receptors in Gliomas

Richard¹,

Kortei

Kampo

2020

ABCMed

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Anti-(glioma surface antigen) monoclonal antibody G-22 recognizes overexpressed CD44 in glioma cells

Cited by 12 publications

References 14 publications

Monoclonal antibody ONS-M21 recognizes integrin α3 in gliomas and medulloblastomas

Monoclonal antibody ONS-M21 recognizes integrin α3 in gliomas and medulloblastomas

Suppression of CD44 expression decreases migration and invasion of human glioma cells

The Pivotal Novel Pathogenic Roles of Hyaluronic Acid and its Receptors in Gliomas

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