Journal: Mediators of Inflammation

Article Title: Lipopolysaccharide-Binding Protein Downregulates Fractalkine through Activation of p38 MAPK and NF- κ B

doi: 10.1155/2017/9734837

Figure Lengend Snippet: SB203580 and SC-514 ameliorated LPS-induced lung injury and inflammation. Hematoxylin and eosin staining was used to determine the LPS-induced lung injury and inflammatory response in the lung tissue of the ARDS rat model. (a) The control group rats (A) had a distinct framework with complete alveolar walls and interstitium without exfiltration. The LPS group rats (B) showed edema, neutrophil infiltration, hemorrhage, moderate bronchiole epithelial desquamation, and minimal hyaline membrane formation. Histotological analysis also show that rats receiving LBPK95A, SB203508, or SC514 injection had substantially less inflammatory cell infiltration, edema, hemorrhage, and thickness of alveolar walls in comparison to rats treated with LPS ((C), (D), and (E), resp.). The photomicrographs are at 400x magnification. ((b) and (c)) The total number of cells, neutrophil number, and neutrophil ratio in BALF were higher in the LPS group than that in the control group and lower in the LPS+LBPK95A, LPS+SB, and LPS+SC groups than that in the LPS group ( p < 0.05, n = 10, in all cases). The MPO of lung tissues and BALF was determined by ELISA. The results show that the level of MPO concentration in the LPS group was significantly higher than that in the control group ( p < 0.05). (d) However, the MPO concentration level in the LPS+LBPK95A, LPS+SB, and LPS+SC groups was significantly lower than that in the LPS group ( p < 0.05 in all cases). (e) The LPS group had a significantly higher W/D ratio than the control group, and the W/D ratio in the LPS+LBPK95A, LPS+SB, and LPS+SC groups were significantly lower than that in the LPS group ( p < 0.05 in all cases). The LPS group had a lower PaO 2 /FiO 2 ratio than the control group. (f) The LPS+LBPK95A, LPS+SB, and LPS+SC groups had higher PaO 2 /FiO 2 ratios than the LPS group ( p < 0.05 in all cases). ∗ represents p < 0.05 when compared with that in the CTL group of rats; # represents p < 0.05 when compared with that in the LPS group of rats; △ represents p < 0.05 when compared with that in the LPS+LBPK95A group of rats ( n = 10).

Article Snippet: The levels of myeloperoxidase activity were measured by the rat MPO ELISA kit (HK105-01, Hycult Biotech, USA) according to the manufacturer's instructions.

Techniques: Staining, Injection, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Journal of Immunology Research

Article Title: CYP4F18-Deficient Neutrophils Exhibit Increased Chemotaxis to Complement Component C5a

doi: 10.1155/2015/250456

Figure Lengend Snippet: Myeloperoxidase (MPO) levels in colonic tissue in DSS colitis. MPO was measured by ELISA in colonic tissue homogenates from wild-type (WT) and Cyp4f18 knockout (KO) mice treated for 9 days with 4% DSS in drinking water (error bars represent SEM, n = 5). Control mice received drinking water without DSS.

Article Snippet: Myeloperoxidase (MPO) was measured using an ELISA kit from Hycult Biotech (HK210).

Techniques: Enzyme-linked Immunosorbent Assay, Knock-Out

Journal: PLoS ONE

Article Title: The Complement Anaphylatoxin C3a Receptor (C3aR) Contributes to the Inflammatory Response in Dextran Sulfate Sodium (DSS)-Induced Colitis in Mice

doi: 10.1371/journal.pone.0062257

Figure Lengend Snippet: C3a/C3a-desArg was measured by enzyme-linked immunosorbent assay (ELISA) in EDTA plasma samples withdrawn by cardiac puncture. Of note, the calculated and depicted values are most likely too high because purified murine C3a had to be used as standard (see . The data shown are means ± SEM from n = 4 (WT, H 2 O), n = 21 (WT, DSS), n = 4 ( C3ar -/- , H 2 O), and n = 21 ( C3ar -/- , DSS) mice for BALB/c, and n = 4 (WT, H 2 O), n = 11 (WT, DSS), n = 4 ( C3ar -/- , H 2 O) and n = 12 ( C3ar -/- , DSS) mice for C57BL/6. n.s., not significant; *, p<0.05.

Article Snippet: To determine myeloperoxidase (MPO), an enzyme-linked immunosorbent assay (ELISA) kit for mouse myeloperoxidase (HyCult Biotechnology, Uden, Netherlands) was used.

Techniques: Enzyme-linked Immunosorbent Assay, Purification

Journal: PLoS ONE

Article Title: The Complement Anaphylatoxin C3a Receptor (C3aR) Contributes to the Inflammatory Response in Dextran Sulfate Sodium (DSS)-Induced Colitis in Mice

doi: 10.1371/journal.pone.0062257

Figure Lengend Snippet: Levels of the granulocyte marker enzyme MPO were measured by enzyme-linked immunosorbent assay (ELISA) in tissue homogenates. The data shown are means ± SEM from n = 7 (WT, H 2 O), n = 21 (WT, DSS), n = 5 ( C3ar -/- , H 2 O), and n = 21 ( C3ar -/- , DSS) mice for BALB/c, and n = 5 (WT, H 2 O), n = 11 (WT, DSS), n = 5 ( C3ar -/- , H 2 O) and n = 12 ( C3ar -/- , DSS) mice for C57BL/6. n.s., not significant; ***, p<0.001.

Article Snippet: To determine myeloperoxidase (MPO), an enzyme-linked immunosorbent assay (ELISA) kit for mouse myeloperoxidase (HyCult Biotechnology, Uden, Netherlands) was used.

Techniques: Marker, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: “Rogue” neutrophil-subset [DEspR+CD11b+/CD66b+] immunotype is an actionable therapeutic target for neutrophilic inflammation-mediated tissue injury – studies in human, macaque and rat LPS-inflammation models

doi: 10.3389/fimmu.2022.1008390

Figure Lengend Snippet: DEspR+ neutrophil-subset roles in LPS-induced tissue injury model. (A) Diagram of LPS-induced multi-organ encephalopathy rat model in hsICH-prone rats. Regular salt-challenge (0.4% NaCl regular rat chow) from embryonic day 0.5 (E0.5) in Dahl Salt-sensitive hypertensive rat inducing hypertension-associated neutrophil/endothelial activation. Sub-endotoxic dose of LPS (1.8 mg/kg IV) was infused in study rats after observation of the 1 st intracerebral hemorrhage event in the age-matched rat cohort around 4m of age (signal ICH ~4m). After LPS was infused, treatment (Tx), either anti-DEspR mAb 10a3 or 6g8 as notated, or mock-treatment (mockTx) saline (vehicle) was given. Treated rats with full recovery were monitored until 35 days (35d). (B) Representative post-mortem images of rat brains after intravascular blood volume replaced with 1X PBS. Left, non-LPS-challenged brain; Middle: anti-DEspR (10a3) mAb treated brain after LPS-infusion. Right: LPS-challenge mock-treated control. (C) Kaplan-Meier survival curve of treated (Tx: 10a3, 1 mg/kg IV, n = 8) vs saline mock-treated (mockTx, n = 9) LPS-challenged hsICH rats. Log Rank Mantel-Cox test: P = 0.0007; hazard ratio (Mantel-Haenszel 10.2, 95% CI of ratio: 2.67 – 39.25). (D-G) Analysis of anti-DEspR mAb target engagement and bioeffects. Study groups are designated (+ or -) per agent (left to right): black open bar = control no LPS and no treatments, black bar = reference control LPS + saline, red bar = LPS + 10a3; red open bar = LPS + 6g8 (D) Minimal to no DEspR+ neutrophils in no-LPS rat control. Compared to LPS + saline control (100% reference), anti-DEspR mAbs 10a3 and 6g8 reduce survival of DEspR+ neutrophils (n = 6 replicates/group), concordant with target engagement and bioeffects in peripheral DEspR+ neutrophils ex vivo . (E) Brain membrane-bound protein levels of mouse-specific immunoglobulin (IgG) in 10a3 or 6g8 mAbs showing brain target engagement, 6g8 > 10a3. (F, G) Analyses of brain non-membrane bound proteins by ELISA detect reduced brain levels of (F) neutrophil-derived myeloperoxidase (MPO) in 10a3 and 6g8-treated rat brains compared to LPS + saline mockTx rat brain, and (G) reduced levels of albumin as a marker of edema in the brain. ng/g brain, nanogram per gram brain; μg/g brain, microgram per gram brain.

Article Snippet: MPO ELISA kit (Hycult Biotech Cat# HK105) and Rat Albumin ELISA kit (AssayPro Cat# ERA2201-1) were used respectively at a 1:10 sample dilution.

Techniques: Activation Assay, Ex Vivo, Enzyme-linked Immunosorbent Assay, Derivative Assay, Marker

Journal: Frontiers in Immunology

Article Title: “Rogue” neutrophil-subset [DEspR+CD11b+/CD66b+] immunotype is an actionable therapeutic target for neutrophilic inflammation-mediated tissue injury – studies in human, macaque and rat LPS-inflammation models

doi: 10.3389/fimmu.2022.1008390

Figure Lengend Snippet: DEspR+ neutrophil-subset roles in LPS-induced tissue injury model. (A) Diagram of LPS-induced multi-organ encephalopathy rat model in hsICH-prone rats. Regular salt-challenge (0.4% NaCl regular rat chow) from embryonic day 0.5 (E0.5) in Dahl Salt-sensitive hypertensive rat inducing hypertension-associated neutrophil/endothelial activation. Sub-endotoxic dose of LPS (1.8 mg/kg IV) was infused in study rats after observation of the 1 st intracerebral hemorrhage event in the age-matched rat cohort around 4m of age (signal ICH ~4m). After LPS was infused, treatment (Tx), either anti-DEspR mAb 10a3 or 6g8 as notated, or mock-treatment (mockTx) saline (vehicle) was given. Treated rats with full recovery were monitored until 35 days (35d). (B) Representative post-mortem images of rat brains after intravascular blood volume replaced with 1X PBS. Left, non-LPS-challenged brain; Middle: anti-DEspR (10a3) mAb treated brain after LPS-infusion. Right: LPS-challenge mock-treated control. (C) Kaplan-Meier survival curve of treated (Tx: 10a3, 1 mg/kg IV, n = 8) vs saline mock-treated (mockTx, n = 9) LPS-challenged hsICH rats. Log Rank Mantel-Cox test: P = 0.0007; hazard ratio (Mantel-Haenszel 10.2, 95% CI of ratio: 2.67 – 39.25). (D-G) Analysis of anti-DEspR mAb target engagement and bioeffects. Study groups are designated (+ or -) per agent (left to right): black open bar = control no LPS and no treatments, black bar = reference control LPS + saline, red bar = LPS + 10a3; red open bar = LPS + 6g8 (D) Minimal to no DEspR+ neutrophils in no-LPS rat control. Compared to LPS + saline control (100% reference), anti-DEspR mAbs 10a3 and 6g8 reduce survival of DEspR+ neutrophils (n = 6 replicates/group), concordant with target engagement and bioeffects in peripheral DEspR+ neutrophils ex vivo . (E) Brain membrane-bound protein levels of mouse-specific immunoglobulin (IgG) in 10a3 or 6g8 mAbs showing brain target engagement, 6g8 > 10a3. (F, G) Analyses of brain non-membrane bound proteins by ELISA detect reduced brain levels of (F) neutrophil-derived myeloperoxidase (MPO) in 10a3 and 6g8-treated rat brains compared to LPS + saline mockTx rat brain, and (G) reduced levels of albumin as a marker of edema in the brain. ng/g brain, nanogram per gram brain; μg/g brain, microgram per gram brain.

Article Snippet: MPO ELISA kit (Hycult Biotech Cat# HK105) and Rat Albumin ELISA kit (AssayPro Cat# ERA2201-1) were used respectively at a 1:10 sample dilution.

Techniques: Activation Assay, Ex Vivo, Enzyme-linked Immunosorbent Assay, Derivative Assay, Marker