Fig. 3. Particulate-containing CD68⁺CD169⁻ macrophages are increased in LLN and exhibit declining function over age.

a. Expression of CD68 and CD169 gated on live, singlet, and CD45⁺CD3⁻cells showing CD68⁺CD169⁻ (black rectangle), CD68-CD169⁺ (red rectangle), and CD68⁺CD169⁺ (blue rectangle) macrophage subsets. b. Differential expression of indicated markers by the three macrophages subsets compiled from LLN shown in a heat map. c. Frequency of macrophage subsets in LLN (n=21) and MLN (n=19). Mixed effects analysis with Tukey’s posttest. Data presented as the mean ± SD. Significance for CD68⁺CD169⁻ LLN vs. MLN: P=0.0004, CD68⁻CD169⁺ LLN vs. MLN: P=<0.0001, CD68⁺CD169⁺ LLN vs. MLN: P=0.9458. d. Frequency of macrophage subsets in LLN (n=21) and MLN (n=19) over age. Linear regression with Pearson correlation (two-tailed). Significance for LLN: CD68⁺CD169⁻ P=0.7447, CD68⁺CD169⁺ P=0.2580; MLN: CD68⁺CD169⁻ P=0.6145, CD68⁺CD169⁺ P=0.1429. Pearson r, *P < 0.033, **P < 0.005 and ***P < 0.001. Data presented as the mean ± 95% CI e. Expression of functional markers CD80/86, CD36, and CD209 by LN CD68⁺CD169⁻ (first and third column) and CD68⁺CD169⁺ (second and fourth column) macrophage subsets in LLNs (n=22, first and second column) and MLNs (n=21, third and fourth columns) over age. Linear regression with Pearson correlation (two-tailed). Significance for LLN: CD68⁺CD169⁻ of % CD80/86⁺ P=0.02075, CD68⁺CD169⁻ of % CD36⁺ P=0.0006202, CD68⁺CD169⁻ of % CD209⁺ P=0.1294, CD68⁺CD169⁺ of % CD80/86⁺ P=0.8834, CD68⁺CD169⁺ of % CD36⁺ P=0.2359, CD68⁺CD169⁺ of % CD209⁺ P=0.7309; MLN: CD68⁺CD169⁻ of % CD80/86⁺ P=0.6393, CD68⁺CD169⁻ of % CD36⁺ P=0.8223, CD68⁺CD169⁻ of % CD209⁺ P=0.3858, CD68⁺CD169⁺ of % CD80/86⁺ P=0.6306, CD68⁺CD169⁺ of % CD36⁺ P=0.8698, CD68⁺CD169⁺ of % CD209⁺ P=0.8307. Pearson r, *P < 0.033, **P < 0.005 and ***P < 0.001. Data presented as the mean ± 95% CI.