1997 Fiscal Year Final Research Report Summary
Function of the gene of Wiskott-Aldrich syndrome
| Project/Area Number |
08457219
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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 | Tohoku University |
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Principal Investigator |
TSUCHIYA Shigeru Tohoku University, Institute of Development, Aging, and Cancer, Associate Professor, 加齢医学研究所, 助教授 (30124605)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIE Hiromi Tohoku University, Institute of Development, Aging, and Cancer, Instructor, 加齢医学研究所, 助手 (60282000)
MINEGISHI Masayoshi Tohoku University, Institute of Development, Aging, and Cancer, Instructor, 加齢医学研究所, 助手 (20211592)
KONNO Tasuke Tohoku University, Institute of Development, Aging, and Cancer, Professor, 加齢医学研究所, 教授 (00004846)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Wiskott-Aldrich syndrome / WASP / gene diagnosis / carrier detection / mouse anti-WASP monoclonal antibody |
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| Research Abstract |
1) Mutation of WASP gene in Wiskott-Aldrich syndrome (WAS)
Mutation of WASP gene was found in 5 cases of WAS.One base (G) deletion in the exon 1 in cases 1 and 2, TCA*TAA change in the exon 2 in case 3, CGA*TGA change in the exon 10 in case 4 and one base deletion (C) in the exon 10 in case 5 were found. These mutations were suggested to give rise to truncated WASP of 63,63,53,320 and 444 amino acids, respectively.
2) Detection of carrier status of WAS
Carriers in family members were detected by PCR-SSCP in case 1 and 2, by Xhol digestion in case 4 and by direct sequencing of PCR product in case 5.
3) Expression of WASPmRNA
WASPmRNA was expressed in mononuclear cells and granulocytes from healthy volunteers by Northern blot analysis. WASPmRNA was not detected in those from the patients. The mRNA was found in human leukemia sell lines in any origin, but not in human fibroblasts.
4) Production of polyclonal and monoclonal antibodies to WASP and detection of WASP by Western blot analysis
Polyclonal and monoclonal (5A5) antibodies were made to WASP peptides and proved to be specific to WASP.Using these antibodies WASP was shown not to express in mononuclear cells of the patients.
5) Flow cytometric detection of cytoplasmic WASP using the 5A5 monoclonal antibody
WASP was clearly negative in cytoplasm of the lymphoblastoid cell lines from the patients as compared in that from normal volunteers. However, strong nonspecific binding interfered the detection of WASP in monocytes, lymphocytes and granulocytes from the patients.
6) Case report of WAS who received bone marrow transplantation and subsequent EBV related B lymphoproliferative disease
The patient was treated by donor lymphocyte transfusion successfully.
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