Journal: Frontiers in immunology

Article Title: Macrophage-Derived Protein S Facilitates Apoptotic Polymorphonuclear Cell Clearance by Resolution Phase Macrophages and Supports Their Reprogramming.

doi: 10.3389/fimmu.2018.00358

Figure Lengend Snippet: Figure 3 | Protein S (PROS1)-deficient macrophages display reduced reprogramming. Macrophages from Pros1fl/fl or LysMCre/+; Pros1fl/fl mice were isolated from peritoneal exudates 66 h after zymosan A injection and incubated for 24 h with vehicle or lipopolysaccharide (LPS, 1 µg/ml). Then, supernatants were collected and analyzed for their content of TNFα [(A), **P = 0.003], IL-6 (B), CCL3 [(C), *P = 0.02], and IL-10 [(D); *P ≤ 0.03] by standard ELISA. (E) IL-10 secretion by untreated or LPS-stimulated control and Pros1-cKO peritoneal macrophages, or by macrophages supplemented with either PROS1 (25 nM), apoptotic cells (AC) or both (AC + PROS1). Results represent the means ± SEM of at least three independent experiments. Two-way ANOVA, *P = 0.02, ***P ≤ 0.0001.

Article Snippet: reagents ELISA kits for mouse TNFα, IL-10, and IL-6 were obtained from Biolegend; a mouse CCL3 detection kit was obtained from R&D Systems.

Techniques: Isolation, Injection, Incubation, Enzyme-linked Immunosorbent Assay, Control

Journal: iScience

Article Title: Metabolic modeling of single bronchoalveolar macrophages reveals regulators of hyperinflammation in COVID-19

doi: 10.1016/j.isci.2022.105319

Figure Lengend Snippet: Glutamine and pyruvate supplementation reduces proinflammatory cytokines and chemokines production in macrophages (A) Violin plot shows glutamine levels in plasma of HCs or patients with COVID-19. Each dot corresponds to an individual patient. (B) Violin plots shows expression levels of SLC1A5 , GLS , and GLUD1 in macrophages from BALF of patients with HCs, mild or COVID-19. (C) Violin plots show relative levels of indicated metabolic fluxes in macrophages from patients with mild and severe COVID-19. (D) GSEA analysis of pyruvate metabolism gene sets in macrophages from patients with severe versus mild COVID-19 . (E) Expression levels of PDHA1 (pyruvate dehydrogenase E1 subunit alpha 1) , PDHB (pyruvate dehydrogenase E1 subunit beta) , and LDHB (lactate dehydrogenase B) in macrophages from BALF of patients with HCs, mild, or severe COVID-19. (F) Violin plots show relative levels of indicated metabolic fluxes in macrophages from patients with mild and severe COVID-19. (G–I) Human primary monocyte-derived macrophages were stimulated with SARS-CoV-2 virus (MOI = 0.1) for 24 h (G). mRNA levels of SLC1A5 , GLUD1 , and GLUL (H) and PDHA1 , PDHB , and LDHB (I) were detected by real-time PCR. Shown are representative data (mock, n = 5; SARS-CoV-2, n = 5) from 3 independent donors with mean values. (J–L) Human primary macrophages cultured in complete medium or medium without glutamine and pyruvate were stimulated with SARS-CoV-2 (J). CCL2 and TFN-α levels in culture supernatant were detected by enzyme-linked immunosorbent assay(ELISA), and CCL3 and IL1B in cells were detected by real-time PCR 24 h after stimulation(K-L). (M) Violin plot shows glucose levels in plasma of HCs or patients with COVID-19. Each dot corresponds to an individual patient. (N–P) Human primary macrophages cultured in complete medium with 2-DG were stimulated with SARS-CoV-2 (N). CCL2 and CCL3 levels in culture supernatant were detected by ELISA, and IL6 and IL1B in cells were detected by real-time PCR 24 h after stimulation (O-P). Shown are representative data (n = 6 for each group) from 3 independent donors with mean values. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Unpaired, two-tailed Student’s t test (for H and I) or one-way ANOVA and Bonferroni’s post hoc test (for K-L and O-P) were performed to compare between groups.

Article Snippet: Human MIP-1alpha (CCL3) ELISA Kit , Peprotech , Cat# 900-K35.

Techniques: Clinical Proteomics, Expressing, Derivative Assay, Virus, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: iScience

Article Title: Metabolic modeling of single bronchoalveolar macrophages reveals regulators of hyperinflammation in COVID-19

doi: 10.1016/j.isci.2022.105319

Figure Lengend Snippet:

Article Snippet: Human MIP-1alpha (CCL3) ELISA Kit , Peprotech , Cat# 900-K35.

Techniques: Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Software, Activity Assay

Journal: iScience

Article Title: Behenic acid alleviates inflammation and insulin resistance in gestational diabetes mellitus by regulating TLR4/NF-κB signaling pathway

doi: 10.1016/j.isci.2024.111019

Figure Lengend Snippet:

Article Snippet: Mouse MIP1α (Macrophage Inflammatory Protein 1 Alpha) ELISA Kit , ELK Biotechnology, China , Cat#ELK1116.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, RNA Extraction, SYBR Green Assay, CCK-8 Assay, Software

Journal: Journal of Translational Medicine

Article Title: Different maturation cocktails provide dendritic cells with different chemoattractive properties

doi: 10.1186/s12967-015-0528-7

Figure Lengend Snippet: Determination of chemokines involved in monocyte chemoattraction. a The expression of different members of the monocyte chemoattractant protein family was evaluated by qPCR. Results are first normalised to the mean expression of two housekeeping genes (GAPDH and HPRT1) and then toward gold standard DC. The relative expression of nine different donor is represented as mean ± SE. *p < 0.05, ns p > 0.05 in the Tukey post hoc test. b The CCL2 and CCL8 concentration of “wash out” supernatants from mature DC seeded in the absence of exogenous stimuli for 24 h were determined by sandwich ELISA. The results are represented as mean ± SE of 8–12 different donors. *p < 0.05, ns p > 0.05 in the Tukey post hoc test.

Article Snippet: Following primers were used: GAPDH (GAGAAGGCTGGGGCTCATTTGC, GGACTGTGGTCATGAGTCCTTCC), HPRT1 (GCTGGATTACATCAAAGCACTG, CTGACCAAGGAAAGCAAAGTCT), CCL2 (CCCCAGTCACCTGCTGTTAT, TGGAATCCTGAACCCACTTC), CCL3L (CTCTGCAACCAGGTCCTCTC, TTTCTGGACCCACTCCTCAC: amplify CCL3L1 and CCL3L3, but not CCL3), CCL4 (TACCATGAAGCTCTGCGTGA, TACCACAAAGTTGCGAGGAA; amplify CCL4, CLL4L1 and CCL4L2), CCL5 (CGCTGTCATCCTCATTGCTA, ACACACTTGGCGGTTCTTTC), CCL8 (CCCTCCAAGATGAAGGTTTC, GAATCCCTGACCCATCTCTC), CCL13 (TGCACTCAACGTCCCATCTA, ACTTCTCCTTTGGGTCAGCA), CCL17 (TTCAAAACCAGGGTGTCTCC, CTGCCCTGCACAGTTACAAA), CCL22 (ACTGCACTCCTGGTTGTCCT, GCTCTTCATTGGCTCAGCTT), CCL23 (TTTGAAACGAACAGCGAGTG, TGTGTCCCTTCACCTTGACA), CXCL8 (TTGCCAAGGAGTGCTAAAGAA, CAGACAGAGCTCTCTTCCATCA), CXCL9 (TGCTGGTTCTGATTGGAGTG, TTTGGCTGACCTGTTTCTCC), CXCL10 (CAACACGTGGACAAAATTGG, TCCAGTCTCAGCACCATGAA), CXCL11 (AGAGGACGCTGTCTTTGCAT, TAAGCCTTGCTTGCTTCGAT), CXCL16 (CTCCTGGCCATCATCTTCAT, AAGCTTCCATTCTTGGCTCA), The chemokine concentrations in the DC “wash out” supernatants were determined by sandwich ELISA for CCL2, CCL3, CCL5, CCL8, CXCL9 and CXCL16 (all from PeproTech) and by flow cytometric plexing for CCL4/MIP-1β, CXCL8/IL8 and CXCL10/IP-10 (CBA multiplexing kit, Becton–Dickinson).

Techniques: Expressing, Concentration Assay, Sandwich ELISA