PMC3169087_01
Immunoprecipitation detection of IgG antibody against ADAMTS13 in plasma. Two micrograms of purified recombinant full-length ADAMTS13 (FL-A13) and the mutant truncated after the CUB domain (delCUB) were incubated with 5 muL of plasma from a well-characterized idiopathic TTP patient and the patient with dengue (this patient) and 40 muL of protein A-Sepharose 4B for 1 hour. The bound IgG-ADAMTS13 or delCUB complex was eluted from the beads after extensive washing by heating the samples at 100 C for 5 minutes with sodium dodecyl sulfate sample buffer and detecting by Western blot with anti-V5 IgG, followed by IRDye800 fluorescent conjugated anti-mouse IgG (1:12,500) and Odyssey image system. Clearly, IgG from both patients recognized FL-A13 and delCUB proteins. The IgG from normal human plasma did not (data not shown).
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F1
The antibody-antigen complexes were determined by immunoprecipitation, followed by Western blot with anti-V5 that recognizes the C-terminal V5-His tag (Fig. 1).
PLT count (x109/L; 
), LDH (IU/L), bilirubin (g/dL), and creatinine (mg/dL) levels during case evolution with plasma exchange.
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F2
TTP by plasma exchange therapy (1.0 plasma volume, daily) due to severity of the case, initiated before laboratorial results confirming DV infection. The time course of clinical and laboratorial findings showed evidence of temporal clinical and laboratorial improvements (Fig. 2).