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. 2015 Jul 30;6:7858. doi: 10.1038/ncomms8858

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(a,b) PMA-stimulated microvessels in the presence of normal human plasma (a) or plasma from a TTP patient with 5% ADAMTS13 activity (b). The left panels in a and b show accumulated vessel wall VWF, and the right panels show zoomed-in views with vessel-wall staining also shown. Green: VWF, red: CD31 and blue: nuclei. (c) Comparison of length range of VWF strands from PMA-stimulated vessels perfused with: PBS, recombinant ADAMTS13, normal plasma or TTP plasma. ***P<0.001 with paired t-test. (d) Accumulation of fluorescent platelets on the vessel wall of stimulated vessels as a function of time with whole blood perfusion in the presence of control IgG (top panels), the ADAMTS13 inhibitory antibody A10 (middle panels), or the combination of A10 and the GPIbα inhibitory antibody AK2 (lower panels). Magenta: CD41a. Flow is from the top left to bottom right. Scale bar, 500 μm. (e) Quantification of adherent platelets on the vessel wall over time in (d). The size and number of thrombi are proportional to the mean fluorescence intensity of labelled platelets. (f,g) VWF-platelet strings/thrombi persists with significant platelet binding near the inlet (f) and less near the outlet (g) in the stimulated vessel after 30 min of whole blood perfusion with inhibited ADAMTS13. Partial blockage occurs with thrombi rich in platelets and VWF (f, left panel). Red blood cells accumulate at sites of platelet-VWF thrombi (f, right panel). Green: VWF, magenta, CD41a and blue: nuclei. (h) Zoomed view showing the structures of persistent VWF fibres bound platelets and red cells. Red: CD41a and green: VWF, white: colocalization of VWF and CD41a. (i) VWF persisted with minimal platelet binding in the stimulated vessel after 30 min of perfusion of blood with both ADAMTS13 and GPIb inhibited. Red: CD31, green: VWF and magenta: CD41a. (j) Blood film of blood treated with ADAMTS13 inhibitory antibody A10 and perfused through a stimulated microvessel showing red cell fragments and very few platelets. Arrows: red cell fragments. N >3 for each condition.