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Distribution of cell‐associated prion protein in normal adult blood determined by flow cytometry
Author(s) -
Barclay G. Robin,
Hope James,
Birkett Christopher R.,
Turner Marc L.
Publication year - 1999
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1999.01789.x
Subject(s) - monoclonal antibody , flow cytometry , microbiology and biotechnology , platelet , peripheral blood mononuclear cell , staining , antibody , blood cell , cell , biology , chemistry , immunology , biochemistry , in vitro , genetics
Leucocyte subpopulations from normally healthy individuals were identified by recognized combinations of fluorochrome‐conjugated antibodies to CD markers and stained by different monoclonal antibodies (MAb) to normal cellular prion protein (PrP C ), including the 3F4 MAb. Cell preparations were examined by three‐colour flow cytometry. All mononuclear leucocyte subpopulations and platelets expressed PrP C , but polymorphonuclear leucocytes and red blood cells expressed little or no PrP C . The amounts of PrP C expressed by the different cells were calculated by comparison to bead standards. Mononuclear leucocytes expressed 3000–4000 molecules of antibody‐reactive PrP C per cell. Resting platelets expressed around 1400 molecules of PrP C per cell, whereas activated platelets expressed around 4800 molecules of PrP C per cell. Extrapolation of these values to the amounts of the various cells in whole blood showed that platelet PrP C accounted for at least 96% of cell‐expressed PrP C in blood. The PrP C on mononuclear cells and platelets was sensitive to enzymatic treatment of cells by proteinase k and phosphatidylinositol‐specific phospholipase C. Certain anti‐PrP C MAbs which showed equivalent intensity of staining to MAb 3F4 on fresh cells showed relative reductions of staining compared to MAb 3F4 on stored cells, indicating possible structural alterations of PrP C under these conditions.