| Project/Area Number |
21591829
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | University of Occupational and Environmental Health, Japan |
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Principal Investigator |
SOU Tetuya 産業医科大学, 医学部, 非常勤医師 (90412642)
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| Co-Investigator(Kenkyū-buntansha) |
HANAGIRI Takeshi 産業医科大学, 医学部, 准教授 (30299614)
TAKENOYAMA Mituhiro 産業医科大学, 医学部, 非常勤講師 (10309966)
BABA Teturou 産業医科大学, 医学部, 助教 (10506348)
安元 公正 産業医科大学, 医学部, 教授 (30150452)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2011: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2009: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Keywords | 悪性胸膜中皮腫 / 細胞障害性T細胞 / 腫瘍浸潤リンパ球 / SEREX法 / Thrombosposqoudin 2 / 腫瘍マーカー / 腫瘍免疫 / Thrombospoudin 2 / 細胞性免疫 / 液性免疫 / 細胞障害性Tリンパ球(CTL) / 腫瘍細胞株 / cDNA発現スクリーニング法 / modified SEREX法 / SCIDマウス血清中ヒト型IgG |
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| Research Abstract |
Malignant pleural mesothelioma(MPM) is difficult to diagnose at an early stage. The present study attempted to identify tumor-associated antigen, which is applicable for surrogate biomarkers of diagnosis or for antigen-targeted therapy for MPM.
Bulk cytotoxic T lymphocytes(CTL) were induced successfully by co-culture of lymphocytes and the autologous MPM cell line L324MSO. Then, CTL clone was established by the limiting dilution method. CTL clone showed cytotoxic activity against L324MSO, but not autologous PHA blast or K562 in an HLA-restricted manner. Using the CTL clone, an antigen-coding gene has been screened using the cDNA expression cloning technique.
On the other hand, tumor-associated antigens were also identified by using antibody derived from tumor-infiltrating B lymphocytes in MPM. Immunoscreening of the cDNA libraries was carried out by serological identification of antigens by a recombinant expression cloning method(SEREX), and 2 antigens were identified as MPM-associated antigens(Gene-X and thrombospondin-2(THBS-2)). Antibodies against Gene-X and THBS-2 in the sera were measured by ELISA method. A total of 124 sera were investigated ; 97 patients were finally diagnosed with MPM and 27 were diagnosed as non-malignant diseases(NM). The serum antibody-titer against THBS-2 was significantly higher in MPM patients than in NM(P<0.01), but there was no difference in the serum antibody-titer against Gene-X. The serum antibody-titer against THBS-2 was a useful non-invasive marker in the diagnosis of MPM. Furthermore, the suppression of Gene-X and THBS-2 expression by transfection with small interfering RNA resulted in suppression of the cell cycle associated gene and apoptosis associated gene, and the growth inhibition of MPM cell lines. These results may indicate that Gene-X and THBS-2 are may be a useful candidate as a target for specific immunotherapy.
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