RETROVIRAL TRANSFER OF P47-PHOX INTO MURINE CELLS AND HUMAN MYELOID CELL LINES
RETROVIRAL TRANSFER OF P47-PHOX INTO MURINE CELLS AND HUMAN MYELOID CELL LINES
批准号:
3809754
负责人:
K LOMAX
金额:
$0.0万
依托单位国家:
美国
项目类别:
财政年份:
--
资助国家:
美国
项目状态:
未结题
起止时间:
至
中文摘要
这个非临床,IIDEA项目的目的是研究可行性
英文摘要
The purpose of this non-clinical, IIDEA project is to study the feasibility
of using retroviral vectors to transfer the gene which is abnormal in
autosomal recessive Chronic Granulomatous Disease(p47-phox) into human
hematopoietic cells. Development of retrovirus producer cell lines capable
of infecting human blood cells will provide the opportunity to study
expression of this protein in precursor and differentiated hematopoietic
cells. A cDNA for p47-phox containing the protein coding-sequence has been
cloned into a retroviral vector known as pLXSN. Two murine producer cell
lines have been established, one producing an ecotropic retrovirus capable
of infecting rodent cells and the other producing amphotropic retrovirus
capable of infecting a wide range of mammalian and avian cells. The
amphotropic virus containing p47-phox in both normal(sense) and
reverse(antisense) orientations was used to infect two leukemia cell lines,
HL-60 and U937. RNA and protein analysis demonstrated retrovirally
transferred p47-phox genes in these cells. In addition, Epstein Barr Virus
transformed B lymphocytes from a normal individual and an AR-CGD patient
were also infected. These lines also appear to have been transduced by the
retroviruses based on protein and RNA analysis.Assays to determine whether
the protein made from the transferred gene is functional are still in
progress. EBV transformed B lymphocyte cell lines derived from AR-CGD
patient cells may be helpful in studies of correction of the defect since
these cells in normals produce a small amount of superoxide and can be
studied at the functional as well as molecular level. Expression of
transferred p47-phox in murine bone marrow and hematopoietic reconstitution
of an irradiated mouse will be necessary before primate and human studies
can proceed. Preliminary experiments indicate that it is possible to
transfer this gene into murine bone marrow.
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CHARACTERIZATION OF THE MURINE HOMOLOG FOR THE HUMAN P47-PHOX GENE
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批准号:3809753
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项目类别:
-
资助金额:$0.0万
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财政年份:--
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负责人:K LOMAX
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依托单位:
海外基金