Journal article Open Access

Generation of HLA-deficient platelets from hematopoietic progenitor cells: GENERATION OF HLA-DEFICIENT PLTs

Figueiredo, Constança; Goudeva, Lilia; Horn, Peter A.; Eiz-Vesper, Britta; Blasczyk, Rainer; Seltsam, Axel


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        <foaf:name>Figueiredo, Constança</foaf:name>
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        <foaf:name>Goudeva, Lilia</foaf:name>
        <foaf:givenName>Lilia</foaf:givenName>
        <foaf:familyName>Goudeva</foaf:familyName>
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        <foaf:name>Horn, Peter A.</foaf:name>
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        <foaf:name>Eiz-Vesper, Britta</foaf:name>
        <foaf:givenName>Britta</foaf:givenName>
        <foaf:familyName>Eiz-Vesper</foaf:familyName>
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        <foaf:name>Blasczyk, Rainer</foaf:name>
        <foaf:givenName>Rainer</foaf:givenName>
        <foaf:familyName>Blasczyk</foaf:familyName>
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        <foaf:name>Seltsam, Axel</foaf:name>
        <foaf:givenName>Axel</foaf:givenName>
        <foaf:familyName>Seltsam</foaf:familyName>
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    <dct:title>Generation of HLA-deficient platelets from hematopoietic progenitor cells: GENERATION OF HLA-DEFICIENT PLTs</dct:title>
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    <dct:issued rdf:datatype="http://www.w3.org/2001/XMLSchema#gYear">2010</dct:issued>
    <dct:issued rdf:datatype="http://www.w3.org/2001/XMLSchema#date">2010-04-23</dct:issued>
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    <dct:description>BACKGROUND: Exposure to allogeneic blood products often leads to the development of human leukocyte antigen (HLA) antibodies. Refractoriness to platelet (PLT) transfusion caused by alloimmunization against HLA Class I antigens constitutes a significant clinical problem. STUDY DESIGN AND METHODS: We developed an RNA interference (RNAi)-based approach to silence the expression of HLA Class I molecules on PLTs derived from CD34+ progenitor cells. A lentiviral-based system was used to express short-hairpin RNA (shRNA) targeting b2-microglobulin (b2m) transcripts in CD34+ progenitor cells. Differentiation to PLTs was performed by incubating progenitor cells in the presence of thrombopoietin and interleukin-3. RESULTS: The transduction of RNAi cassettes containing the sequences for shRNAs targeting b2m caused up to 85% reduction of progenitor cells HLA Class I antigen expression, which was maintained in the culture-derived PLTs. The HLA-deficient PLTs derived from HLA-silenced CD34+ cells proved to be fully functional in in vitro tests when compared to peripheral blood–derived PLTs. CONCLUSIONS: Our data show that in vitro generating HLA Class I–deficient PLTs from hematopoietic progenitor cells prove to be feasible. As malignancy risks associated with insertional mutagenesis are not to be expected in anucleated PLTs, provision of HLA-deficient PLTs from large-scale production units may become reality in the management of patients suffering from PLT transfusion refractoriness.</dct:description>
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