1989 Fiscal Year Final Research Report Summary
Models of immunotherapy using monoclonal antibodies against human small cell lung cancer.
| Project/Area Number |
63480210
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Respiratory organ internal medicine
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| Research Institution | Osaka University |
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Principal Investigator |
KAWASE Ichiro Osaka University, Medical School, Associate, 医学部, 助手 (10161324)
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| Co-Investigator(Kenkyū-buntansha) |
INOUE Tamotsu Osaka University, Faculty of Health and Sport Sciences, Associate, 健康体育部, 助手 (00203240)
TANIO Yoshiro Osaka University, Medical School, Associate, 医学部, 助手 (50197521)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Small Cell Lung Cancer / Cluster 1 Antigen / Monoclonal Antibody / Antibody-dependent Cellular Cytotoxicity / Immunotoxin |
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| Research Abstract |
New cell lines of small cell lung cancer (SCLC), OSI, OS2 and OS3, were established from specimens of untreated primary tumors biopsied by diagnostic bronchofiberscopy. Clinical responses of these tumors were well correlated with in vitro sensitivity of their respective cell lines to chemotherapeutic drugs and irradiation. Biological and biochemical analysis revealed that OS2, possessing neuroendocrine features, is the classic type and the other 2 lines are the variant type of SCLC.
Two murine monoclonal antibodies (MoAb) against OS1 were obtained and named ITK-2 and -3. Both of the 2 MoAbs are IgGl. ELISA, immunostaining and FACS analysis revealed that these 2 MoAbs recognize the cluster 1 antigen of SCLC.
ELISA after the treatment of OSI with enzymes and periodate showed that the antigenic determinant recognized by ITK-2 is glycoprotein. Its molecular weight was shown to be 140kd in SDS-PAGE following immunoprecipitation of ^<125>I-labeled OS1 cell membrane with ITK-2.
Competitive binding inhibition assay using ^<125>I-labeled ITK-2 revealed that ITK-2 binds to the different site from that those of MOC-1 and NKH-1, both of which recognize the cluster 1 antigen. On the other hand, the binding of ^<125>I-labeled ITK-2 was significantly enhanced by the pretreatment of SCLC cells with ITK-3.
4-h ^<51>Cr-release assay showed that ITK-2 induced antibody-dependent cellular cytotoxicity against cells of both SCLC and neuroblastoma lines, when LAK cells were used as effector cells.
ITK-2-ricin A chain conjugate was constructed. The immunotoxin specifically inhibited protein synthesis, cell proliferation and colony formation of SCLC cells in vitro. Furthermore, the growth of SCLC tumor in BALB/c nude mice was significantly inhibited by intratumoral injections of the immunotoxin.
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