| Project/Area Number |
05670743
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Dermatology
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| Research Institution | Kitasato University School of Medicine |
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Principal Investigator |
MASUZAWA Mikio Kitasato Univ. Sch. of Med. Associate Prof., 医学部, 助教授 (30129267)
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| Co-Investigator(Kenkyū-buntansha) |
MIYATA Toshiko Kitasato Univ. Sch. of Med. Assistant, 医学部, 助手 (30260855)
AMANO Takafumi Kitasato Univ. Sch. of Med. Assistant, 医学部, 助手 (30212567)
FUJITA Yuhsuke Kitasato Univ. Sch. of Med. Assistant, 医学部, 助手 (20238588)
FUJIMURA Takao Kitasato Univ. Sch. of Med. Assistant, 医学部, 助手 (50209087)
天野 隆文 北里大学, 医学部, 助手 (50255288)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1994: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1993: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Malignant hemangioendo-thwlioma / bispecific antibody / helper / killer cells / rIL-2 / LAK / adoptive immunotherapy / 悪性血管内皮細胞 / ペルパー / キラー |
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| Research Abstract |
LAK adoptive immunotherapy is recognized as the one and the only effective therapy for malignant hemangioendothelioma (MHE) that is an extremely fatal neoplasm. To advance this therapy, we examined basic studies of bispecific antibody (Ab) therapy using helper/killer cells.
We established a human malignant endothelial cell line (ISO-HAS) as tumor antigen (Ag) to make monoclonal antibody (monoAb) and as target cell for cytotoxic assay. It was derived from MHE arising on the scalp and expressed several oncogenes and mutated suppessive oncogenes. Using ISO-HAS,we obtained some Abs responsive to both ISO-HAS and normal endothelial cells (EC), but no specific Ab against ISO-HAS.
CD4^+T cells were separated from PBMC by magnetic cell separation system (MACS). We cultured CD4^+killer cells by bi-stimulation of rIL-2 and anti-CD3 Ab. Discontinuous stimulation by immobilized anti-CD3 Ab was needed to culture them massively. By RT-PCR methods, we observed the expression of mRNA of IL-2 and gamma IFN in CD4^+killer cells. These cells alone showed about 70% cytotoxicity against ISO-HAS in high E/T ratio (100/1).
Our results suggested that tumor-specific Ag of MHE is too weak to make a monoAb. Moreover, to show high cytotoxic activity in low E/T ratio, CD4^+killer cells need a bridging Ab to make contact with tumor cells. Therefore, we are now making a bispecific Ab using ABs to CD3 and CD31 which is preserved on MHE as well as normal EC.We will present the final report including the results of our studies using this bispecific Ab.
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