Figure 3.

CD4+T cells from subjects with interstitial lung abnormalities (ILA) loss the KLRG1 expression compared with old control group. Peripheral mononuclear cells were prepared to flow cytometry and western blot. First, live cells were identified, the gate of CD3+CD4+was delimited, and then the expression of CD27, CD28, CD57, KLRG1 and PD1 was evaluated by percentage and mean fluorescence intensity (MFI) (panels A and B, respectively). Panel C: Total T cells were obtained using a Pan T kit and then CD4+T cells were enriched. The expression of KLRG1 was evaluated in the fraction of CD4+T cells by western blot under non-reducing condition. Panel D: Band densities were normalised against GAPDH by densitometry analysis and results are shown in relative units of concentration using IMAGEJ software; bars show mean±SD. In panels A and B, graphs show individual value mean±SD from: Young: n=12, old: n=13, old+ILA: n=10. Western blot was performed in two subjects per group. *p<0.05 **p<0.01. Statistical analysis was performed with multiple t-tests and a Holm-Sidak method was used to adjustment for multiple comparisons; asterisks indicate comparison of the three groups. When the asterisk is on a line, indicates, the comparison between old control group and old+ILA. CCR7, C-C chemokine receptor 7; KLRG1, killer-cell lectin-like receptor G1; PD-1, programmed cell death 1.