Triglycerides, C-reactive protein, and interleukin-6 were natural log-trans- formed. Model A: adjusted for age, race, and sex. Model B: adjusted for age, race, sex, smoking, diabetes mellitus, hypertension, systolic blood pressure, and BMI. Model C: adjusted for age, race, sex, smoking, diabetes mellitus, hy- pertension, systolic blood pressure, BMI, LDL cholesterol, HDL cholesterol, C- reactive protein, interleukin-6, and fibrinogen. In all models, the estimates for age, race, and sex were adjusted only for the 2 remaining variables. *P<0.0001.

2. Top Genomewide Association Results for Soluble CD14 in European Americans and Blacks

Number SNPs Top SNP in Minor Minor Allele Effect Size (SE) Population Chromosome With P<5×10−8 Region Position Hg18 Candidate Genes Allele Frequency [ng/mL sCD14] P Value European-Americans 5q31 144 rs5744441 139997031 HBEGF, SLC4A9, ANKHD1, G 0.22 −101 (10) 2.98×10–23 EIF4EBP3, SRA1, APBB3, SLC35A4, CD14, IK, WDR55, TMCO6, DND1, HARS, HARS2, NDUFA2,ZMAT2, VTRNA1 Black 5q31 1 rs778584 139985396 CD14 T 0.31 128 (24) 5.5×10–9 European-Americans 1q24 19 rs1063412 170677590 PIGC A 0.43 −48 (8.5) 1.40×10–8

Black

SNP indicates single-nucleotide polymorphism.

rs5744441 (LD r2=1.0). After conditioning on rs5744455, probability values for 3 additional SNPs in the CD14 region were significant at the Bonferroni-adjusted threshold of P<0.003: rs2569191 and rs778584 (which are in strong LD; r2=0.97) and rs4914, suggesting the possibility of allelic heterogeneity at the CD14 locus. Together with rs5744455, rs4914 (a synonymous coding variant) and rs778584 define 4 common CD14 EA haplotypes (Table 3). Compared with ref- erent haplotype 1, haplotype 4 (tagged by the minor allele of rs5744455) was associated with lower sCD14, whereas haplo- type 3 (tagged by the minor allele of rs4914) and haplotype 2 (tagged by the minor allele of rs778584) were each associated with higher sCD14 levels.

were attributed to noncardiovascular causes. The relation- ships between sCD14 and these outcomes are shown in Table 4. When minimally adjusted for age, sex, and race, baseline sCD14 was associated with increased risk of MI, CHD, stroke, and mortality. Additional adjustment for traditional risk fac- tors and measures of subclinical vascular disease attenuated these associations, though the associations with CHD, stroke, and mortality remained significant. Moreover, the association of sCD14 with mortality and stroke (but not MI or CHD) was independent of the other studied biomarkers (CRP, IL-6, fibrin- ogen). In the fully adjusted model 4, sCD14 was associated similarly with death because of cardiovascular causes (hazard ratio=1.13; 95%CI=1.05–1.22; P=0.001) and death because of noncardiovascular causes (hazard ratio=1.11; 95%CI=1.04– 1.18; P=0.001). Table II in the online-only Data Supplement shows the association of each inflammation biomarker with mortality in multivariable models, including all the inflamma- tion biomarkers. In these models, sCD14 and IL-6 were each independently associated with mortality, whereas CRP and fibrinogen were not. When analyzed categorically, there was a graded, linear increase in mortality risk with each succes- sive sCD14 quintile. Compared with the bottom quintile, the hazard ratios (95% CIs) for the second, third, fourth, and fifth quintile were 1.00 (0.87–1.14), 1.02 (0.89–1.16), 1.12 (0.98– 1.27), and 1.27 (1.11–1.45), respectively. The hazard ratios comparing top with bottom quintiles for cardiovascular death and noncardiovascular were 1.26 (95% CI 1.01–1.57; P=0.04) and 1.32 (95% CI 1.11–1.57; P=0.002), respectively.

In blacks, several other SNPs on the IBC array had more significant P values than the index SNP from the black GWAS rs778584. The lead SNP from the IBC analysis was rs5744451 (MAF=0.09), which was associated with 279±36 ng/mL lower sCD14 (P=1.3×10–14). rs5744451 is located in the 5ʹ flanking region of CD14. This SNP was not successfully imputed and hence not included in the GWAS analysis. After conditioning on rs5744451, 3 additional CD14 SNPs (rs5744455, rs2569191, and rs778584) had P<0.003. Haplotype analysis confirmed that rs5744451 tags a second CD14-lowering haplotype that is com- mon among blacks but rare among EA (Table 3).

Relationship of sCD14 to Incident CVD and Mortality

Among the 5462 EA and black participants with measured at the CHS baseline examination, there were incident MI, 1547 incident CHD, 945 incident strokes, 3820 deaths during the 20-year follow-up period. Of the deaths, 1519 were attributed to cardiovascular causes and

Among the sCD14-associated SNPs identified in our GWAS, there was no association with risk of mortality in either ethnicity of CHS (Table III in the online-only Data Supplement). In CHS, the minor alleles of CD14 rs4914

3. Association of CD14 haplotypes With Soluble CD14 in European Americans and Blacks

CD14 SNP European Americans Blacks Haplotype rs778584 rs4914 rs5744455 rs5744451 Freq β (SE) P Value Freq β (SE) P Value H1 C C G A 0.17 Referent  0.44 Referent … H2 T C G A 0.49 67 (11) 6.15e-08 0.31 51 (25) 0.04 H3 C G G A 0.12 87 (15) 3.23e-08 0.07 26 (44) 0.55 H4 C C A A 0.21 −46 (12) 0.0002 0.08 −93 (40) 0.02 H5 C C G C <0.01 NA NA 0.09 −262 (37) 3.2e–013

SNP indicates single-nucleotide polymorphism; NA, not applicable; and EA, European The global P value for CD14 haplotype association with sCD14 in EA=6.65e–31. The global P value for CD14 haplotype association with sCD14 in blacks=1.28e–12.

4. Estimated Hazard Ratios for Association Between 1 SD Unit Increase in Soluble CD14 and Incident CVD Events and