Identification of macrophage activation-related biomarkers in obese type 2 diabetes that may be indicative of enhanced respiratory risk in COVID-19

Abu Saleh Md Moin, Thozhukat Sathyapalan, Ilhame Diboun, Stephen L Atkin, Alexandra E Butler, Abu Saleh Md Moin, Thozhukat Sathyapalan, Ilhame Diboun, Stephen L Atkin, Alexandra E Butler

Abstract

Hyperactivation of the immune system through obesity and diabetes may enhance infection severity complicated by Acute Respiratory Distress Syndrome (ARDS). The objective was to determine the circulatory biomarkers for macrophage activation at baseline and after serum glucose normalization in obese type 2 diabetes (OT2D) subjects. A case-controlled interventional pilot study in OT2D (n = 23) and control subjects (n = 23). OT2D subjects underwent hyperinsulinemic clamp to normalize serum glucose. Plasma macrophage-related proteins were determined using Slow Off-rate Modified Aptamer-scan plasma protein measurement at baseline (control and OT2D subjects) and after 1-h of insulin clamp (OT2D subjects only). Basal M1 macrophage activation was characterized by elevated levels of M1 macrophage-specific surface proteins, CD80 and CD38, and cytokines or chemokines (CXCL1, CXCL5, RANTES) released by activated M1 macrophages. Two potent M1 macrophage activation markers, CXCL9 and CXCL10, were decreased in OT2D. Activated M2 macrophages were characterized by elevated levels of plasma CD163, TFGβ-1, MMP7 and MMP9 in OT2D. Conventional mediators of both M1 and M2 macrophage activation markers (IFN-γ, IL-4, IL-13) were not altered. No changes were observed in plasma levels of M1/M2 macrophage activation markers in OT2D in response to acute normalization of glycemia. In the basal state, macrophage activation markers are elevated, and these reflect the expression of circulatory cytokines, chemokines, growth factors and matrix metalloproteinases in obese individuals with type 2 diabetes, that were not changed by glucose normalisation. These differences could potentially predispose diabetic individuals to increased infection severity complicated by ARDS. Clinical trial reg. no: NCT03102801; registration date April 6, 2017.

Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Circulatory levels of M1 macrophage activation markers in the steady state of obese type 2 diabetes (OT2D). Plasma LBP (A), TLR4 (B), CD80 (C) and CD38 (D) levels in control subjects (black circles) and obese subjects with T2D (blue circles). Basal level of plasma LBP was higher (85,311 ± 1453 vs 91,747 ± 3048 RFU of LBP, OT2D vs control, p < 0.05) but TLR4 was unchanged in OT2D (235 ± 11 vs 254 ± 13 RFU of TLR4, OT2D vs control, p = ns). Basal levels of plasma M1 macrophage surface markers CD38 and CD80 were also higher in OT2D (714 ± 24 vs 626 ± 21 RFU of CD80, OT2D vs control, p < 0.01; 441 ± 15 vs 408 ± 7 RFU of CD38, OT2D vs control, p < 0.05). Acute normalization of glycemia had no effect on levels of LBP, CD80 or CD38 in T2D subjects (A-D). Data were presented as mean ± SEM. *, p < 0.05, **, p < 0.01.
Figure 2
Figure 2
Circulatory levels of cytokines and chemokines released from activated M1 macrophages in the steady state of obese type 2 diabetes (OT2D). Plasma CXCL1 (A), CXCL5 (B), CCL5 (RANTES) (C), CXCL9 (D) and CXCL10 (E) levels in control subjects (black circles) and obese subjects with T2D (blue circles). Basal level of plasma CXCL1, CXCL5 and RANTES was higher in OT2D compared to control (2487 ± 177 vs 2060 ± 67 RFU of CXCL1, OT2D vs control, p < 0.05; 543 ± 17 vs 500 ± 10 RFU of CXCL5, OT2D vs control, p < 0.05; 36,961 ± 5692 vs 18,162 ± 2393 RFU of RANTES, OT2D vs control, p < 0.01). Basal levels of plasma CXCL9 and CXCL10 were lower in T2D compared to control (455 ± 26 vs 722 ± 113 RFU of CXCL9, and 1825 ± 122 vs 2706 ± 352 RFU of CXCL10, OT2D vs control, p < 0.05). Acute normalization of glycemia had no effect on levels of CXCL1. CXCL5, RANTES, CXCL9 and CXCL10 in T2D subjects (A-E). Data were presented as mean ± SEM. *, p < 0.05, **, p < 0.01.
Figure 3
Figure 3
Circulatory levels of activated M2 macrophage markers in the steady state of obese type 2 diabetes (OT2D). Plasma TGF-β1 (A), CD163 (B), MMP7 (C) and MMP9 (D) levels in control subjects (black circles) and obese subjects with T2D (blue circles). Basal levels of plasma TGF-β1, CD163, MMP7 and MMP9 were higher in OT2D compared to control (1123 ± 72 vs 933 ± 27 RFU of TGF-β1, OT2D vs control, p < 0.01; 2748 ± 256 vs 2297 ± 124 RFU of CD163, OT2D vs control, p < 0.05). Two matrix metalloproteinases MMP7 and MMP9 were also increased in OT2D (1242 ± 93 vs 1005 ± 42 RFU of MMP7, OT2D vs control, p < 0.05 and 30,193 ± 3746 vs 19,532 ± 1562 RFU of MMP9, OT2D vs control, p < 0.01). Acute normalization of glycemia had no effect on levels of LBP, CD80 or CD38 in T2D subjects (A-D). Data were presented as mean ± SEM. *, p < 0.05, **, p < 0.01.
Figure 4
Figure 4
Circulatory levels of adipose tissue macrophage (ATM) markers in the steady state of obese type 2 diabetes (OT2D). Plasma level of MCP-1 (A), MIF (B) and netrin-1 (C) in control subjects (black circles) and obese subjects with T2D (blue circles). Basal level of plasma MCP-1 did not alter in OT2D compared to control (698 ± 49 vs 760 ± 73 RFU of MCP-1, OT2D vs control, p = ns). Basal level of plasma MIF and netrin-1was increased in OT2D (1387 ± 223 vs 1007 ± 29 RFU of MIF-1 and 558 ± 30 vs 478 ± 19 RFU of netrin-1, OT2D vs control, p < 0.05). Acute normalization of glycemia had no effect on levels of LBP, CD80 or CD38 in T2D subjects (A-C). Data were presented as mean ± SEM. *, p < 0.05.

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