Antiplatelet Effects of Clopidogrel Vs Aspirin in Virologically Controlled HIV

Figures

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  Figure 1. Clinical Trial and Experimental Design Schematic of clinical trial. (A) Study site visit and platelet phenotype assessment at (T1) enrollment, (T2) randomization, and (T3) follow-up at day 14. (B) Freshly isolated platelets (n = 2 per group) coincubated on HUVECs for 2 hours at T2 and T3. Isolated mRNA from HUVECS underwent NanoString transcriptomic analysis (n = 594), whereas a separate validation cohort (n = 11 to 14 per group) underwent platelet–HUVEC coincubation experiments with subsequent quantitative polymerase chain reaction (of HUVEC transcriptome expression) at study end (T3). HIV = human Immunodeficiency virus; HUVEC = human umbilical vein endothelial cell.

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  Figure 2. AA- and ADP-Induced Platelet Aggregation Are Inhibited After Aspirin and Clopidogrel Treatment Percent platelet aggregation using light transmission aggregometry. (A) Representation of platelet aggregation studies and (B) percent aggregation at 300 seconds in response to 1,600 μmol/L arachidonic acid by study assignment. (C) Representation of platelet aggregation studies and (D) percent aggregation at 300 seconds in response to 5.0 μmol/L of ADP by study assignment. n = ∼22 per group (aspirin/clopidogrel), n = ∼11 per group (control); ∗∗P < 0.01, ∗∗∗P < 0.001 by Wilcoxon signed-rank test. Bar graphs indicate median with IQR. AA = arachidonic acid; ADP = adenosine diphosphate; asa = aspirin; bl = baseline; clo = clopidogrel; Ctrl = control; fu = follow-up.

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  Figure 3. Platelet P-Selectin and PAC-1 Expression Is Reduced After Aspirin and Clopidogrel Treatment Platelet flow-cytometry (expressed as mean fluorescence units [MFI]). Resting (A) and (B) thrombin (0.05 U/mL) stimulated platelet p-selectin (CD62p+, CD42b+) expression in each treatment group. Resting (C), arachidonic acid (D, 0.60 μmol/L), ADP (E, 0.1 μmol/L), and epinephrine (F, 0.4 μmol/L) stimulated PAC-1 (CD42b+ PAC-1+) expression. All stimulated treatments occurred over 5 minutes. ∗∗P < 0.01, ∗∗∗P < 0.001 by Wilcoxon signed rank test. Bar graphs indicate median with IQR. Abbreviations as in Figure 2.

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  Figure 4. Platelet-Treated Composite Endothelial Transcript Expression Is Reduced After Clopidogrel Freshly isolated platelets at study randomization and follow-up (n = 2 per study assignment group) were coincubated on human umbilical vein endothelial cells (HUVECS) × 2 hours and the HUVEC transcriptome assessed using the NanoString immunology panel (594 genes). (A) Venn diagram displacing directionality and change in all measurable HUVEC transcripts by study assignment (∼246 genes). (B) Average change, expressed as percentage, in the composite HUVEC transcriptome by study assignment. (C) Top 5 down-regulated canonical pathways in the clopidogrel treatment arm vs aspirin and control arms (by Ingenuity Pathway Analysis). (D) Normalized (to control) fold change in HUVEC transcripts obtained from C by study assignment. ∗∗∗P < 0.001 (analysis of covariance and Student’s t-test for posttreatment scores). iNOS = inducible nitric oxide synthase; other abbreviations as in Figure 2.