Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Eosinophil levels in eAT, blood, bone marrow, liver, and intestine of rIL5 injected mice . (A ) Male C57BL/6J mice were fed either chow or HFD for 8 weeks and simultaneously IP injected twice weekly with rIL5 protein. (B) Flow cytometry gating strategy for eosinophils (CD45 + , live, F4/80 lo , CD11b lo , SiglecF + ) and macrophages (CD45 + , live, F4/80 hi , CD11b hi , SiglecF − ). (C) Percent eAT eosinophils negatively correlate with body weight (slope = −0.96, R 2 = 0.75). (D) Treatment with rIL5 elevates eAT eosinophils in HFD-fed mice, compared to saline and vehicle controls, back to chow-fed levels [n = 4–11]. (E) No difference in percent eosinophils in blood, bone marrow, or liver of rIL5 treated mice, but intestine does show an increase [n = 2–8]. Data are shown as means ± SEM. eAT = epididymal adipose tissue; HFD = high fat diet. Key: chow = hashed bars; HFD = open bars; saline = white; vehicle = grey; rIL5 = black. * P < 0.05, compared to saline; *** P < 0.0005, compared to saline; ˆˆ P < 0.005, compared to vehicle.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques: Injection, Flow Cytometry, Saline

Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Inflammatory profile of obese eAT with elevated eosinophils . (A ) Representative 40× images of eAT from HFD-fed obese mice treated with saline, vehicle, or rIL5. Macrophages (red, F4/80) are seen in abundance in all groups, whereas eosinophils (green, SiglecF) are increased in rIL5 treated mice. Magnified view shows the donut-shaped nucleus ( * ) of eosinophils juxtaposed to the spherical nucleus (#) of macrophages [n = 3]. (B) Treatment with rIL5 does not alter percent eAT macrophages or (C) protein expression of pro-inflammatory marker MHCII [n = 6–11]. (D) Gene expression analysis of macrophage marker, Adgre1 (F4/80), and an array of macrophage polarization genes were not statistically different between groups in eAT [n = 6–8]. Data are shown as means ± SEM. eAT = epididymal adipose tissue, HFD = high fat diet. Key: saline = white; vehicle = grey; rIL5 = black.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques: Saline, Expressing, Marker, Gene Expression

Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Weight gain, body composition, and glucose tolerance in mice with elevated AT eosinophils . (A) HFD-fed mice gained more weight over 8 weeks compared to chow, with no difference in (B) total weight gained between vehicle or rIL5 treated mice [n = 4]. HFD-fed vehicle and rIL5 treated mice have equally altered total fat and lean mass by (C) percent or (D) grams at study completion [n = 4–12]. (E) HFD-fed mice have increased body, eAT, and sAT weight, but no further difference with rIL5 [n = 4]. (F) Glucose tolerance is impaired by HFD-feeding, but not rescued by rIL5 treatment [n = 4]. Data are shown as means ± SEM. AT = adipose tissue; HFD = high fat diet; eAT = epididymal AT; sAT = subcutaneous AT. Key: chow = hashed bars, dotted lines; HFD = open bars, solid lines; vehicle = grey; rIL5 = black. * P < 0.05, compared to chow; ** P < 0.005, compared to chow; *** P < 0.0005, compared to chow; **** P < 0.0001, compared to chow; ˆ P < 0.005, compared to vehicle.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques:

Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Triglyceride tolerance in mice with elevated AT eosinophils . Fasted HFD-fed mice treated with vehicle or rIL5 for 8 weeks were given a bolus of TGs (i.e. olive oil) and showed no difference between groups in (A) plasma TG [n = 8] (B) plasma FFA [n = 4] (C) blood glucose [n = 8] (D) or plasma cholesterol [n = 8] over the course of 5 h. Data are shown as means ± SEM. HFD = high fat diet; TG = triglycerides; FFA = free fatty acids. Key: vehicle = grey; rIL5 = black.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques: Clinical Proteomics

Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Metabolic parameters of rIL5 treated mice during a mixed-meal test . HFD-fed mice treated with vehicle or rIL5 for 8 weeks were fasted and then allowed access to HFD ad libitum for 3 h. (A) Both groups consumed the same amount of food [n = 6–7]. (B) A spike in blood glucose occurred at 0.5 h following refeeding with no difference between groups [n = 6–7]. (C) Insulin also rose at 0.5 h and then held relatively steady in both groups [n = 6–7]. Plasma (D) TG [n = 6–7], (E) FFA [n = 4], and (F) cholesterol [n = 5–7] did not vary between groups upon refeeding. Data are shown as means ± SEM. HFD = high fat diet; TG = triglycerides; FFA = free fatty acids. Key: vehicle = grey; rIL5 = black.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques: Clinical Proteomics

Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Insulin signaling in eAT with elevated eosinophils . (A) Representative western blots of eAT showed an increase in the ratio pAKT/AKT upon insulin stimulation in HFD-fed mice that received vehicle or rIL5 for 8 weeks [n = 3–4]. β-Actin is shown as a protein loading control. (B) Quantification of western blots revealed no difference in the eAT insulin signaling response via pAKT/AKT between vehicle and rIL5 treated mice. Data are shown as means ± SEM. eAT = epididymal adipose tissue; HFD = high fat diet. Key: vehicle = grey; rIL5 = black; insulin injected = dotted bars; no-insulin saline control injection = open bars. * P < 0.05, compared to no insulin stimulation.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques: Western Blot, Control, Injection, Saline

Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Cold challenge: Energy balance in mice with elevated AT eosinophils . HFD-fed mice that previously received vehicle or rIL5 for 8 weeks were individually housed in metabolic cages to measure an array of physiological parameters for 2 days at RT and 2 days at 4 °C [n = 8]. (A) Average daily body weight did not vary between vehicle and rIL5 treated mice. (B) Average daily food intake and (C) average daily movement were increased in the dark cycle compared to light cycle, but with no difference between vehicle and rIL5. (D) The respiratory quotient oscillated between light and dark cycles to equal degrees in vehicle and rIL5 treated mice, both during RT and the 4 °C cold challenge; indicating both groups had the same energy substrate utilization. (E) Energy expenditure increased upon 4 °C cold challenge to maintain body heat, but did not vary between vehicle and rIL5 treated mice. Both treatment groups lost the same amount of (F) body mass, (G) fat mass, and (H) lean mass. Data are shown as means ± SEM. eAT = epididymal adipose tissue; sAT = subcutaneous AT. RT = room temperature, set at 21 °C; HFD = high fat diet. Key: vehicle = grey bars/lines; rIL5 = black bars/lines.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques:

Journal: Molecular Metabolism

Article Title: Elevating adipose eosinophils in obese mice to physiologically normal levels does not rescue metabolic impairments

doi: 10.1016/j.molmet.2017.12.004

Figure Lengend Snippet: Cold challenge: Beiging capacity of white AT with elevated eosinophils . AT of HFD-fed mice that received vehicle or rIL5 for 8 weeks was examined after 2 days of RT or 4 °C cold exposure. After cold exposure, the presence of increased eosinophils was verified in (A) sAT and (C) eAT of rIL5 treated mice compared to vehicle controls [n = 5–7]. Cold exposure alone was able to increase eosinophils in sAT but not eAT. (B) Ucp1 gene expression in sAT was increased ∼307-fold in the cold compared to RT, with no further increase in rIL5 treated mice [n = 4–5]. (D) Ucp1 gene expression in eAT trended towards an increase under cold conditions, but no difference was observed between vehicle and rIL5 treated mice [n = 5–7]. Data are shown as means ± SEM. eAT = epididymal adipose tissue; sAT = subcutaneous AT; RT = room temperature, set at 21 °C; HFD = high fat diet; Ucp1 = uncoupling protein 1. Key: vehicle = grey bars; rIL5 = black bars.

Article Snippet: During the entire 8-week period, mice also received 50 ng recombinant IL5 (rIL5; R&D Systems, Minneapolis, MN) protein (in 0.001% BSA as carrier protein) by intraperitoneal (IP) injection twice per week to elevate eosinophils.

Techniques: Gene Expression