T-Helper 17 Cell/Regulatory T-Cell Imbalance in COPD Combined with T2DM Patients

Figures & data

Table 1 Demographics and Clinical Characteristics in the Each Group of Subjects

Variables	Healthy Control Group	COPD Group	T2DM Group	COPD+T2DM Group	P-value
Subjects (No.)	30	36	35	32
Age (year)	65.82±9.37	70.24±8.14	68.17±8.47	66.30±10.72	0.189
Gender (Male/female)	21/9	28/8	24/11	25/7	0.725
BMI (kg/m^2)	24.34±3.82	23.83±4.95	26.43±4.21	24.86±3.57	0.062
Smoking rate, n (%)	6 (20%)	15 (41.67%)	8 (22.86%)	13 (40.63%)	0.368
Fasting blood glucose (mmol/L)	4.39±1.12	4.43±0.99	8.96±3.57^**##	7.87±3.02^**##	<0.001
2h postprandial blood glucose (mmol/L)	6.24±2.17	6.83±2.46	14.52±4.98^**##	13.61±4.55^**##	<0.001
HbA1c (%)	5.64±1.72	6.08±1.65	8.34±2.02^**##	8.70±2.36^**##	<0.001
FEV1/FVC%	89.25±9.43	51.57±10.61**	85.64±11.32^##	43.24±8.06^**##ΔΔ	<0.001
FEV1% predicted	83.12±8.46	50.87±8.24**	79.05±9.15^##	42.37±9.30^**##ΔΔ	<0.001

Notes: Data are presented as mean ± SD. **P<0.01 compared with the healthy control group; ^##P<0.01 compared with the COPD group; ^ΔΔP<0.01 compared with the T2DM group.

Abbreviations: BMI, body mass index; FEV1, forced expiratory volume in 1 second; FVC, forced vital capacity.

Table 2 Measurement Indicators in the Each Group of Subjects

	Healthy Control Group	COPD Group	T2DM Group	COPD+T2DM Group	F-value	P-value
Th17 (CD4^+T%)	0.568±0.325	1.365±0.673**	1.230±0.801**	2.127±0.917^**##ΔΔ	24.553	<0.001
Treg (CD4^+T%)	3.955±0.937	3.704±1.164	1.835±0.872^**##	1.822±0.819^**##	12.085	<0.001
Th17/Treg	0.172±0.093	0.506±0.247**	0.659±0.281**	1.157±0.563^**##ΔΔ	45.502	<0.001
RORγt	1.000 ± 0.000	10.800 ± 1.175**	9.684 ± 1.505**	25.670 ±5.117^**##ΔΔ	14.11	< 0.001
Foxp3	1.000 ± 0.000	0.718 ± 0.048**	0.748 ± 0.014**	0.435 ± 0.059^**##ΔΔ	73.28	< 0.001
IL-17A (pg/mL)	6.894±0.538	9.995±1.679**	10.194±1.868**	12.519±1.754^**##ΔΔ	26.892	<0.001
IL-17F (pg/mL)	2.606±0.907	4.120±1.296**	4.529±1.319**	6.356±1.508^**##ΔΔ	45.034	<0.001
IL-21 (pg/mL)	3.267±1.267	5.111±1.093**	4.339±2.087*	7.993±2.198^**##ΔΔ	43.193	<0.001
IL-23 (pg/mL)	8.507±2.701	16.558±4.652**	14.534±5.507**	20.040±3.956^**##ΔΔ	37.595	<0.001
IL-6 (pg/mL)	19.257±3.562	25.248±4.601**	24.307±4.291**	33.514±5.313^**##ΔΔ	19.344	<0.001
IL-l0 (pg/mL)	10.876±6.809	7.868±3.916*	8.084±3.219**	4.913±1.241^**##ΔΔ	10.417	<0.001
TGF-β1(pg/mL)	25.498±5.021	20.704±5.807**	19.008±5.477**	15.142±4.065^**##ΔΔ	21.414	<0.001

Notes: Data are presented as mean ± SD. *P<0.05, **P<0.01 compared with the healthy control group; ^##P<0.01 compared with the COPD group; ^ΔΔP<0.01 compared with the T2DM group. GAPDH was used as internal reference to detect the statistical results of RORγt and Foxp3.

Figure 1 Representative flow cytometric dot-plots of peripheral blood Th17 cells in the each group of subjects. (A) is Healthy control; (B) is COPD patient; (C) is T2DM patient; (D) is COPD+T2DM patient. The percentage of positive cells is shown in each panel.

Figure 2 Representative flow cytometric dot-plots of peripheral blood Tregs cells in the each group of subjects. (A) is Healthy control; (B) is COPD patient; (C) is T2DM patient; (D) is COPD+T2DM patient. The percentage of positive cells is shown in each panel.

Figure 3 Imbalance of circulating Th17 and Tregs in COPD, T2DM and COPD combined with T2DM patients. (A) Comparisons of Th17 and Treg cell percentages in peripheral blood from healthy controls, COPD, T2DM and COPD combined with T2DM patients. (B) Comparisons of Th17 cell percentages in the 4 groups. (C) Comparisons of Treg cell percentages in the 4 groups. (D) Comparisons of the ratios of Th17/Treg in the 4 groups. The data are represented as the mean ± SD; a value of *P <0.05 was considered statistically significant.

Figure 4 The relative levels of Foxp3 and RORγt mRNA transcripts to the control glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in PBMCs from healthy controls, COPD, T2DM and COPD combined with T2DM patients. (A and C) Comparisons of RORγt in the 4 groups. (B and D) Comparisons of Foxp3 in the 4 groups. The data are represented as the mean ± SD; a value of **P <0.01 was considered statistically significant. COPD (C group), T2DM (D group) and COPD combined with T2DM (C+D group).

Figure 5 Disordered homeostasis of serum IL-17A, IL-17F, IL-21, IL-23, IL-10, TGF-β1 and IL-6 in COPD, T2DM and COPD combined with T2DM patients. Levels of inflammatory cytokines were quantified by ELISA in the 4 groups (A) IL-17A, (B) IL-17F, (C) IL-21, (D) IL-23, (E) IL-10, (F) TGF-β1, (G) IL-6. The data are represented as the mean ± SD; a value of *P <0.05 or **P <0.01 was considered statistically significant. COPD (C group), T2DM (D group) and COPD combined with T2DM (C+D group).