Plasma and platelet CFH content in response to plasma transfusion in a constitutive CFH-null patient treated with plasma transfusion. Immunoblots (nonreduced 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis) of equivalent plasma aliquots (left) and platelet lysates (right) from a normal person (lane “normal”) and aHUS patient 1 lacking constitutive expression of CFH (but capable of expressing FHL-1) tested before therapeutic plasma transfusion (lane “before”), after treatment (lane “after”), and after treatment at days 1, 7, and 14 (corresponding lanes; transfusion interval was 14 days). Loading was equivalent to 0.7 μL of plasma and lysate from 107 platelets per lane. Immunoblots were initially probed with a rabbit polyclonal antibody capable of detecting CFH and FHL-1, and subsequently with antibodies specific for coagulation FXII for the plasma blot, and β-actin for the platelet lysate blot as loading controls. CFH is deficient in plasma and platelets before (lane “before”) and on day 14 (lane “day 14”) compared with a normal person (lane “normal”), whereas FHL-1 levels were unchanged. CFH levels were highest right after transfusion (lane “after”) and then declined over the following 14 days (lanes “day 1,” “day 7,” and “day 14”) in both plasma (left panel) and platelets (right panel). A vertical line has been inserted in the right panel (“Platelet”) to indicate a repositioned gel lane.
