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Blood, 16 April 2009, Vol. 113, No. 16, pp. 3838-3844. Prepublished online as a Blood First Edition Paper on January 9, 2009; DOI 10.1182/blood-2008-09-178475.
Submitted September 10, 2008
Department of Immunology and Transfusion Medicine, University Hospital of North Norway, Tromso, Norway * Corresponding author; email: tor.brynjar.stuge{at}fagmed.uit.no.
T cell responses have been implicated in the development of HPA-1a-induced neonatal alloimmune thrombocytopenia (NAIT). However, HPA-1a-specific T cells have neither been isolated nor characterized. Here, we aimed to determine whether HPA-1a-specific T cells could be isolated from HPA-1a-immunized women. In the present study, PBMCs from an HPA-1a-alloimmunized woman were cultured for weeks in the presence of HPA-1a peptide, labelled with CFSE, and assayed for antigen-specific proliferation. Individual proliferating cells were isolated by fluorescence-activated cell sorting and expanded in culture. Antigen specificity and HLA restriction were determined by cytokine secretion (ELISPOT) and proliferation assays. Several CD3+CD4+ T cell clones were isolated, that proliferated and secreted cytokines in response to HPA-1a peptide. Two of these clones have been established in long term culture in our laboratory. Both of these recognize synthetic as well as naturally processed HPA-1a antigen, and the recognition is restricted by the MHC molecule HLA-DRB3*0101, that is strongly associated with NAIT. These HPA-1a-specific T cell clones represent unambiguous evidence for the association of T cell responses with NAIT, and will serve as unique tools to elucidate the cellular immune response that may result in NAIT.
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