2004 Fiscal Year Final Research Report Summary
INDUCTION OF VIRUS-SPECIFIC CYTOTOXIC T LYMPHOCYTES FROM CORD BLOOD LYMPHOCYTES.
| Project/Area Number |
13470168
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Tokai University |
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Principal Investigator |
KATO Shunichi Tokai University, School of Medicine, Professor, 医学部, 教授 (70096212)
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| Co-Investigator(Kenkyū-buntansha) |
HAGIHARA Masao Tokai University, School of Medicine, Assistant Professor, 医学部, 講師 (00208422)
HATTORI Kinya Tokai University, School of Medicine, Research Assistant, 医学部, 助手 (50276820)
INOUE Hiroyasu Tokai University, School of Medicine, Research Assistant, 医学部, 助手 (40366000)
TANAKA Kazuo Tokai University, school of medicine, Associate Professor, 医学部, 助教授 (50236569)
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| Project Period (FY) |
2001 – 2004
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| Keywords | cytomegalovirus / CTL / Dendritic cells / immunotherapy |
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| Research Abstract |
A : Results of studies using human cord blood & periopheral blood
Cytomegalovirus (CMV) infection is a severe complication in post-stem cell transplanted (SCT) patients. In the present study, we have developed a culture system to expand anti-CMV specific cytotoxic killer T cells (CTL) using peripheral blood from normal healthy volunteers.
1. Ant-CMV CIL was successfully generated either from sero-positive or -negative persons using crude antigen-pulsed monocytes-derived dendritic cells.
2. M A2402 or 0201 restricted CMV-tetramers were used to isolate anti-CMV CTL which were then cultured with those peptides. In either method, killer activities together with antigen-specificities were confirmed by in vitro 51Cr release assays.
3. In SCT recipients, anti-CMV cellular immunity was successfully monitored by anti-CMV HLA tetramers in vivo.
B : Results of studies using mice
In order to confirm therapeutic efficacy of the adoptive transfer of cord-blood-derived cytomegalovirus (CMV)-specific CTL, murine cytomegalovirus, CMV's murine counterpart, was utilized. In this study, BALB/c mice (H-2d) were infected with MCMV In the preliminary experiments, we established the real time PCR method to measure a small amount of MCMV, and made H-2Ld/MCMV-IE tetramers, which could detect MCMV-specific CTLs. Using these techniques, we obtained several results showing below
1. MCMV-specific CTL could be maintained under the condition where MCMV-genome was undetectable. This result suggest that the adoptively transferred CMV-specific CTLs, which are in vitro generated, can be maintained in a CMV sero-negative recipient
2. Indigenous bacterial flora plays a role in maintaining and expanding of virus-specific CTL
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