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ANALYSIS OF ANTIGENICITY OF MUC-1 MUCIN AS A TUMOR ANTIGEN AND POSSIBILITY OF THIS MOLECULE FOR THE USE OF COMBINED ANTI-CANCER IMMUNOTHERAPY WITH GENE TRANSFER

Research Project

Project/Area Number 08671356
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General surgery
Research InstitutionSHIGA UNIVERSITY OF MEDICAL SCIENCE

Principal Investigator

INOUE Shuhei  SHIGA UNIVERSITY OF MEDICAL SCIENCE, SECOND DEPARTMENT OF SURGERY,ASSOCIATED PROFESSOR, 医学部, 助手 (30273402)

Co-Investigator(Kenkyū-buntansha) KATO Horofumi  SHIGA UNIVERSITY OF MEDICAL SCIENCE,SECOND DEPARTMENT OF SURGERY,ASSOCIATED PROF, 医学部, 講師 (20111974)
OKADA Yoshio  SHIGA UNIVERSITY OF MEDICAL SCIENCE,PRINCIPAL, 医学部, 学長 (10106825)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsMUC-1 mucin / Tumor antigenicity / Anti-tumor immunity / Gene transfer / Anti-cancer immunotherapy / Preparation for clinical intervention
Research Abstract

(1)Establishment of a MUC-1 transduced cancer cell line and analysis of its antigenicity as a tumor antigen
AMUC-1 transduced cell line was established by transfection of MUC-1 cDNA into breast cancer cell line obtained from a cancer patient. To study the antigenicity of the tranduced cell line, cytotoxic activity of peripheral blood lymphocytes (PBL) against the cell line was examined. A strong lytic activity wa boserved.
(2)Induction of cytotoxic T lymphocytes (CTL) specific for MUC-1 mucin and characterization of them
MUC-1 specific CTL were induced by co-culture of PBL from cancer patients with the MUC-1 tranduced cell line in the presence of cytokines. The induced CTL showed CD3/CD4 surface markers and TCRalphabeta chain on the surface by FACS.
(3)Analysis of antigenic determinants and specificity of the mucin molecule recognized by the CTL
The induced CTL were studied for the cytotoxic activity against the MUC-1 transduced cell line by cytotoxicity assay. Although a strong activity was seen against the MUC-1-expressing cell line, no activity was against the parent cell line. Adding anti-MUC-1 antibodies showed an inhibitory effect on the cytotoxicity observed.
(4)Effect of B7 and IL-12 on enhancement of CTL function
Apparent propagation of the CTL was induced via B7 molecule expressed on NZK-muc. On the other hand, CTL induced using stimulators without B7 expression showed negligible propagation. IL-12 supplement resulted in differentiation of CTL into CD8 positive T lymphocytes.
(5)Targetting MUC-1 mucin on cancer cells for combined anti-cancer immunotherapy with gene transfer technique
To enhance the CTL function, transfection of cytokine gene into the CTL is in process. Retrovirus expression vectors inserted with TNF-alpha and interferon-beta cDNA have been prepared.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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