Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Human peripheral blood CD14+ monocytes , iXCells , Cat # 10HU-008.

Techniques: Recombinant, Isolation, Enzyme-linked Immunosorbent Assay, Cell Isolation, Software, Imaging

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Bone Marrow Mesenchymal Stem Cell-Derived Dermcidin-Containing Migrasomes enhance LC3-Associated Phagocytosis of Pulmonary Macrophages and Protect against Post-Stroke Pneumonia.

doi: 10.1002/advs.202206432

Figure Lengend Snippet: Figure 7. DCD is beneficial to AIS recovery and DCD-containing BM-MSC-derived migrasome effectively promotes phagocytosis of macrophages. A– G) Peripheral blood of AIS patients (acute phase, 0–3d after disease onset, n = 16) and healthy controls (HC, n = 8) were collected. (A) Plasma DCD concentration was assessed with ELISA. *p < 0.05, compared with HC by Student’s t-test (mean ± standard deviation). (B) Correlation of clinic parameters and plasma DCD concentration was assessed with Spearman correlation analysis and Point-biserial correlations. *p < 0.05. DM, diabetes mellitus, CHD, coronary heart disease. (C) Representative images of the magnetic resonance diffusion weighted imaging (MR-DWI) of AIS patients with low plasma DCD concentration (DCD ≤3.33 ng ml−1) or high plasma DCD concentration (DCD > 3.33 ng ml−1). (D) Association between plasma DCD concentration with infarct scale was estimated with Spearman correlation analysis. (E) Association between plasma DCD concentration with delta NIHSS (NIHSS at 7d minus NIHSS at 1d) was estimated with Spearman correlation analysis. (F) Representative images of the chest Computed Tomography (CT) of AIS patients with low plasma DCD concentration (DCD ≤3.39 ng ml−1, median of the cohort) or high plasma DCD concentration (DCD > 3.39 ng ml−1, median of the cohort). (G) Pie charts showing the occurrence of post-stroke pneumonia in AIS patients with low and high plasma DCD concentrations. H) DCD (1 ng ml−1), PBS-migrasomes (PBS-M, 50 μg ml−1) or E. Coli-migrasomes (E. Coli-M, 50 μg ml−1) labeled with Dil (red) were treated to BMDM (15 min). Immunostaining of WGA (green) and DCD (withe) in migrasome-treated BMDM was performed. Experiments were repeated for three times. I,J) BMDM were first pre-stimulated with DCD (1 ng ml−1), PBS-M (50 μg ml−1), or E. Coli-M (50 μg ml−1) for overnight then treated with E. Coli (E. Coli : BMDM = 20:1, 1 h). Phagocytic efficiency of BMDM to GFP expressing E. Coli was assessed with flow cytometry (I) and immunostaining (J). Experiments were repeated three times. **p < 0.01, compared with PBS-treated group by one-way ANOVA (mean ± standard deviation).

Article Snippet: Human Monocyte Enrichment and Macrophage Differentiation: Mononucleus cells were isolated from peripheral blood of healthy adults (age = 18–40y) with human peripheral blood monocyte isolation Solution kit (Solarbio, P8680).

Techniques: Derivative Assay, Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Imaging, Computed Tomography, Labeling, Immunostaining, Expressing, Cytometry

Journal: Journal of Virology

Article Title: Human Cytomegalovirus Decreases Major Histocompatibility Complex Class II by Regulating Class II Transactivator Transcript Levels in a Myeloid Cell Line

doi: 10.1128/JVI.01901-19

Figure Lengend Snippet: HCMV downregulates surface expression of MHC class II in a myeloid progenitor cell line. (A) Flow cytometry analysis of unstained Kasumi-3 cells or cells stained for HLA-DR, HLA-DQ, and HLA-DP. (B) Flow cytometry scatter plot of HCMV-infected (72 hpi) and uninfected cells showing the relationship between mCherry (a marker of HCMV infection) and HLA-DR. (C) Histograms of HCMV-infected (mCherry) and uninfected Kasumi-3 cells stained for surface HLA-DR at 72 and 120 h postinfection. (D) Bar graph of the geometric mean (GM) fluorescence values for the samples used in the assay whose results are presented in panel C, displayed as a percentage of the value for the uninfected sample. (E) Geometric mean fluorescence values from uninfected or HCMV-infected CD14+ human peripheral blood monocytes (HPBM) at 72 hpi. Values are averages from a minimum of three independent experiments. *, P < 0.05.

Article Snippet: Human peripheral blood monocytes (catalog number 6906K-50A; Cell Applications Inc.) were maintained in the manufacturer’s supplied human blood cell culture medium.

Techniques: Expressing, Flow Cytometry, Staining, Infection, Marker, Fluorescence

Journal: Molecular Metabolism

Article Title: Activated macrophages control human adipocyte mitochondrial bioenergetics via secreted factors

doi: 10.1016/j.molmet.2017.07.008

Figure Lengend Snippet: Effects of LPS/IFNγ- and IL10/TGFβ-activated primary MΦ-CM on bioenergetics of human SGBS and primary adipocytes . Primary human CD14 + monocytes from 4 healthy donors were cultured, differentiated to macrophages (naïve MΦs) and activated with LPS/IFNγ or IL10/TGFβ as described in . (A) Morphological changes detected by bright field microscopy (one representative experiment shown). (B) mRNA expression of CD163 and CD40 analyzed with ΔΔCT method using naïve MΦs as Calibrator (n = 4). (C) CD163 and CD40 mRNA expression presented as ratio. (B, C) *p < 0.05 vs. LPS/IFNγ-activated MΦ, ***p < 0.001 vs. LPS/IFNγ-activated MΦ, ###p < 0.001 vs. naïve MΦs. (D) mRNA expression of UQCRC2 and NDUFB8 analyzed with ΔΔCT method using naïve THP1 macrophages as Calibrator (n = 4). ( E–G) SGBS adipocytes were treated with either cell-free control media, LPS/IFNγ-activated or IL10/TGFβ-activated MΦ-CM collected from 3 different donors for 48 h as described in and analyzed as described in Figure 4 , Figure 5 . Data are normalized to cell number (DNA content) and are the mean + SEM of 5 independent experiments each performed in 2–8 wells condition. OCR and ECAR traces vs time are shown in ( Figure S4B and C ). (E) ATP-linked respiration of SGBS adipocytes after treatment for 48 h with indicated MΦ-CM. (F) ATP-linked respiration of SGBS adipocytes presented as relative change in OCR in percent of cell-free control. (G) ECARs due to glycolysis of SGBS adipocytes after treatment for 48 h with indicated MΦ-CM. (H – J) Human primary subcutaneous adipocytes were treated with either cell-free control media, LPS/IFNγ-activated or IL10/TGFβ-activated MΦ-CM from 3 different donors as described in . Bioenergetic profile was analyzed as described in Figure 4 , Figure 5 . Data are presented as fold change to the mean rates of cell-free control for each donor and are the mean + SEM of experiments from 3 adipocyte donors performed in 6–8 wells per condition and donor. OCR and ECAR traces vs time are shown in ( Figure S4E and F ) (H) ATP-linked respiration of human primary adipocytes after treatment for 48 h with indicated MΦ-CM. (I) ATP-linked respiration of human primary adipocytes presented as relative change in OCR in percent of cell-free control. (J) ECARs due to glycolysis of human primary adipocytes after treatment for 48 h with indicated MΦ-CM. (E)–(J) *p < 0.05 vs. LPS/IFNγ-activated MΦ-CM; **p < 0.01 vs. LPS/IFNγ-activated MΦ-CM; ***p < 0.001 vs. LPS/IFNγ-activated MΦ-CM; #p < 0.05 vs. cell-free control media.

Article Snippet: Human CD14 + blood monocytes from 4 healthy donors were purchased from Zenbio (Research Triangle Park, NC, USA) and differentiated with 10 ng/ml CSF1 for 7 days to adherent macrophages (MΦ) before same stimulation as for THP1 cells was performed.

Techniques: Cell Culture, Microscopy, Expressing, Control

Journal: Stem Cell Research & Therapy

Article Title: Ixmyelocel-T, an expanded multicellular therapy, contains a unique population of M2-like macrophages

doi: 10.1186/scrt345

Figure Lengend Snippet: Ixmyelocel-T macrophages express M2 macrophage surface receptors. (A) Ixmyelocel-T stained with the M2 macrophage markers CD163 and CD206.

Article Snippet: For the generation of M1 and M2 macrophages, human peripheral blood monocytes from healthy donors were purchased (AllCells LLC., Alameda, USA).

Techniques: Staining

Journal: Stem Cell Research & Therapy

Article Title: Ixmyelocel-T, an expanded multicellular therapy, contains a unique population of M2-like macrophages

doi: 10.1186/scrt345

Figure Lengend Snippet: Ixmyelocel-T macrophages express M2 markers. Quantitative real-time PCR gene expression analysis of M1 and M2 macrophage markers within the ixmyelocel-T, M1, and M2 macrophages normalized to GAPDH (n ≥ 4). Relative expression of (A) PPARγ, (B) CD206, (C) CD163, (D) CD204, (E) SR-B1, (F) MERTK, (G) CCR7, (H) TNFα, (I) IL-1β, and (J) TGFβ in M1, M2, and IXT macrophages. Values are presented as mean ± SEM relative to IXT, * P <0.05 , ** P < 0.01, *** P < 0.001 vs M1, # P <0.05 , ## P < 0.01, ### P < 0.001 vs M2. PPAR-γ, peroxisome proliferator-activated receptor gamma; TGFβ, transforming growth factor beta.

Article Snippet: For the generation of M1 and M2 macrophages, human peripheral blood monocytes from healthy donors were purchased (AllCells LLC., Alameda, USA).

Techniques: Real-time Polymerase Chain Reaction, Gene Expression, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Ixmyelocel-T, an expanded multicellular therapy, contains a unique population of M2-like macrophages

doi: 10.1186/scrt345

Figure Lengend Snippet: Ixmyelocel-T macrophages remain anti-inflammatory after inflammatory challenge. (A) TNFα and (B) IL-12 were quantified in M1, M2, and ixmyelocel-T supernatants treated with and without 0.1 μg/mL LPS (n = 4–7). Values are presented as mean ± SEM relative to control, * P <0.001 vs M1. # P <0.001 vs M2. & P <0.05 vs IXT. ^ P <0.05, ^^ P <0.001 vs M1 + LPS. % P <0.001 vs M2 + LPS. LPS, lipopolysaccharide; SEM = standard error of the mean.

Article Snippet: For the generation of M1 and M2 macrophages, human peripheral blood monocytes from healthy donors were purchased (AllCells LLC., Alameda, USA).

Techniques: Control