Study of induction and evaluation of the specific immunological response by peptides derived from lung cancer associated antigens

2001 Fiscal Year Final Research Report Summary

• Project/Area Number: 11670587
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Respiratory organ internal medicine
• Research Institution: Nagoya City University
• Principal Investigator: KAWAGUCHI Haruhiko Nagoya City University Medical School Research Associate, 医学部, 助手 (90305532)
• Co-Investigator(Kenkyū-buntansha): OONO Tomoyosi Nagoya City University Medical School Research Associate, 医学部, 助手 (90264712) ; UEDA Ryuzo Nagoya City University Medical School Professor, 医学部, 教授 (20142169)
• Project Period (FY): 1999 – 2000
• Keywords: peptide / specific cytotoxicity / flow cytometry

Research Abstract

In this study, we introduced the lung cancer specific cytotoxcic immunoresponse by peptide derived from both HER2 and MAGES. As antigen presenting cell we used dendritic cells (DCs) that were introduced from peripheral blood mononuclear cells (PBMCs) of normal volunteers with either HLA A2402 or HLA 0201 after co culturing with IL-4 and GM-CFS. We generated peptide specific cytotoxic T cells (CTLs) by incubating auto PBMGs with DCs in the presence low dose IL-2 and ll-7 in addition to peptides. To generate highly purified CTLs, we selected CTLs by CD8 (+) negative selection using magnetic beas. For standard evaluation of cytotoxic activity against HER2 (+) lung cancer cell lines, we measured radioactivity in the supernatant after lysis of target cells. We observed both peptides derived from HER2 and MAGE3 specific CTL activity by this method and to explore more simple evaluation method, we investigated the usefulness of flow cytometric method (FACS) as an indicator of CTL activity. We stained CTLs with both anti IFN-gamma and anti CDS monoclonal antibodies labeled either FITC or PE and examined the frequency of double positive cells by FACS. The result obtained by FACS method was significantly correlated with the standard radioactivity against target cells. Because the number of double positive cells was a little few, further improvement in detection for CTLs by FACS is needed to apply this method for indicator of peptide specific CTL activity.