| 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)
清水 崇史 東海大学, 医学部, 助手 (00338782)
保田 由喜治 東海大学, 医学部, 助手 (60265874)
矢部 普正 東海大学, 医学部, 講師 (70220217)
矢部 みはる 東海大学, 医学部, 講師 (40172514)
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| Project Period (FY) |
2001 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2004: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥3,700,000 (Direct Cost: ¥3,700,000)
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| Keywords | cytomegalovirus / CTL / Dendritic cells / immunotherapy / CMV-CTL / テトラマー / HLA / 臍帯血 / DC / CTL / ウイルスの特異的免疫 / ウイルス特異的免疫 / 臍帯血移植 / ヘルペスウイルス / リンパ球幼若化反応 / real time PCR / CD8+CD11b- |
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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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