Lineage-specific expression of CYBB in leukocytes
| Project/Area Number |
12670994
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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 |
Hematology
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| Research Institution | Institute of Tropical Medicine, Nagasaki University |
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Principal Investigator |
NAKAMURA Michio Institute of Tropical Medicine, Nagasaki University, Department of Host-Defense Biochemistry, Professor, 熱帯医学研究所, 教授 (30091276)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKIM Shoichi Institute of Tropical Medicine, Nagasaki University, Department of Host-Defense Biochemistry, Assistant, 熱帯医学研究所, 助手 (40253695)
KUMATORI Atsushi Institute of Tropical Medicine, Nagasaki University, Department of Host-Defense Biochemistry, Instructor, 熱帯医学研究所, 講師 (60244092)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2001: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | CYBB / gp91phox / flavocytochrome / PU.1 / HAF-1 / CGD / Pu box / STAT-1α / LRF-L / GATA / 慢性肉芽腫症 |
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| Research Abstract |
(1) Roles of PU.1 and HAF-1 for the expression of CYBB
Single base substitutions were systematically induced to the CYBB promoter and examined their effects on reporter activities and binding affinities to PU.1 and HAF-1. Data suggested that PU.1 is principal and HAF-1 supplementary. These also implied new CGDs and supplied ideally modified promoter sequences for maximal expression of the gene in the CGD phagocytes.
(2) A new cw-elemement for the activation of CYBB
We discovered a new GATA-binding site on the CYBB promoter. This site is essential for the expression of the gene promoted by PU.1 and GATA-1.
(3) A role of IRF-4 in B lymphocytes
IRF-4 was suggested to inhibit CYBB that may account for the low expression of gp91phox in B lymphocytes.
(4) Trials for GATA-1 gene transfection to phagocytes of the X-CGD patient with an abnormal CYBB promoter
For making gp91phox-deficient neutrophils/monocytes to gp91phox-normal eosinophils, we tried to transfect GATA-1 gene to the peripheral blood monocytes of a patient with X-linked bp-53T CGD patient, Efficiencies of liposomal and adenoviral transfections were inefficient but activated the gene nonspecifically suggesting a potential of the gene to be activated endogenously.
(5) Identification of the epitope for 7D5, a monoclonal antibody raised against flavocytochrome b558
7D5 epitope was molecular-biologically located to the extracellular peptide conformation of gp91phox. The antibody is now undoubtedly available for diagnosis of X-CGD and monitoring the efficiency of the gene ttransfer to CGD patients.
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Report
(3 results)
Research Products
(13 results)
