Journal: Biochimica et Biophysica Acta. Molecular Basis of Disease

Article Title: Single cell RNA-seq resolution revealed CCR1 + /SELL + /XAF + CD14 monocytes mediated vascular endothelial cell injuries in Kawasaki disease and COVID-19

doi: 10.1016/j.bbadis.2023.166707

Figure Lengend Snippet: The differential activation of CD14 and CD16 monocytes responding to endothelial and epithelial dysfunction. The profile scores of molecules mediating the interplays of classic monocytes with vascular endothelial cells (A-B) and non-classic monocytes with alveolar epithelial cells (C—D). E. Immunostaining for CDH5 and γH2AX in coronary artery cryo-section between KD patient and healthy donor, indicating endothelial injuries in KD. F. Expression level of CCR1 , DYSF , SELL , LMNB1, and XAF1 in CD14 classical monocytes among five groups. G. UMAP projection of CCR1 , DYSF , SELL , LMNB1, and XAF1 positive cells, respectively . H. UMAP projection of CCR1 , SELL and XAF1 triple positive monocytes among five groups. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, Wilcoxon rank-sum test, adjusted for Bonferroni post hoc test, had been applied.

Article Snippet: Cell smears fixated in 4 % PFA were blocked in blocking buffer (PBS containing 1 % BSA, 0.1 % Triton X-100, 5 % goat serum) for 1 h at room temperature, followed by incubation with rabbit anti-Dysferlin (1:200, Abcam, cat # ab124684), rabbit anti-XAF1 (1:200, Abcam cat #ab2254) antibodies and mouse anti-CD14 (1:200, Abcam cat #ab181470) diluted in PBS containing 1 % BSA, 5 % goat serum at 4 °C overnight.

Techniques: Activation Assay, Immunostaining, Expressing

Journal: Biochimica et Biophysica Acta. Molecular Basis of Disease

Article Title: Single cell RNA-seq resolution revealed CCR1 + /SELL + /XAF + CD14 monocytes mediated vascular endothelial cell injuries in Kawasaki disease and COVID-19

doi: 10.1016/j.bbadis.2023.166707

Figure Lengend Snippet: SELL+/CCR1+/XAF1+ CD14 monocytes enhanced the adhesion and damages to endothelial cells. A. Flow cytometry identified the higher percentages and median fluorescence intensity of SELL, CCR1, and LMNB1 in KD and COV. B. Immunostaining for CD14 and DYSF, XAF1 in isolated PBMCs. C. The ratio of DYSF positive CD14 monocytes in total classic monocytes among KD, COV, FLU, and healthy donors. And the median fluorescence intensity of XAF1 in CD14 monocytes among KD, FLU, and healthy donors. D-E. THP-1 had been stained with Calcein AM and transfected siRNAs of CCR1 , DYSF , SELL , LMNB1 , and XAF1 before co-culture with HUVECs. Then the numbers of adhesion THP-1 with HUVECs were counted by every 20× field view. F. The expressions of TNFa and IL6 in HUVECs, and the ratio of γH2AX + cells in HUVECs after co-cultured with THP-1, which had been transfected with siRNAs of CCR1 , DYSF , SELL , LMNB1 , and XAF1 . G. The inhibition of SELL , CCR1 and XAF1 together in THP-1 significantly reduced the adhesion between monocytes and endothelial cells. H. Collaborated inhibiton of SELL , CCR1 and XAF1 in THP-1 decreased the expression of IL6 and TNF-α in HUVECs with lower ratio of rH2AX+ cells after co-culture with THP-1. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, Two-way analysis of variance with Bonferroni post hoc test was performed to analyze data. Bar, 100 μm.

Article Snippet: Cell smears fixated in 4 % PFA were blocked in blocking buffer (PBS containing 1 % BSA, 0.1 % Triton X-100, 5 % goat serum) for 1 h at room temperature, followed by incubation with rabbit anti-Dysferlin (1:200, Abcam, cat # ab124684), rabbit anti-XAF1 (1:200, Abcam cat #ab2254) antibodies and mouse anti-CD14 (1:200, Abcam cat #ab181470) diluted in PBS containing 1 % BSA, 5 % goat serum at 4 °C overnight.

Techniques: Flow Cytometry, Fluorescence, Immunostaining, Isolation, Staining, Transfection, Co-Culture Assay, Cell Culture, Inhibition, Expressing

Journal: Scientific Reports

Article Title: Quantity of alcohol drinking positively correlates with serum levels of endotoxin and markers of monocyte activation

doi: 10.1038/s41598-017-04669-7

Figure Lengend Snippet: Box plots showed the levels of sCD14 and sCD163 in the serum of controls and excessive drinkers (ED)

Article Snippet: Commercially available enzyme-linked immunosorbent assays were used according to the manufacturers’ protocols for measuring sCD14 (R&D System, Cat#DC140, Minneapolis, MN), and sCD163 (MyBioSource, Cat # MBS728094, San Diego, CA).

Techniques:

Journal: Scientific Reports

Article Title: Quantity of alcohol drinking positively correlates with serum levels of endotoxin and markers of monocyte activation

doi: 10.1038/s41598-017-04669-7

Figure Lengend Snippet: ( A , B ) Peripheral blood mononuclear cells (PBMCs) were isolated from healthy controls ( A and B ) and pre-treated with ethanol ranging from 6.25 mM–50 mM (1.4 mg–10 mg%) followed by treatment with LPS (10 ng/ml for 6 hrs). Ethanol primes PBMCs for LPS-induced inflammatory responses, as indicated by the increase in the levels of sCD14 and sCD163 in the supernatant in the dose dependent manner. ( C , D ) PBMCs were isolated from excessive drinkers and subjected to the stimulation with LPS (10 ng/ml for 6 hrs). sCD14 and sCD163 were significantly increased in supernatants of LPS stimulated PMBCs from subjects with EAU compared to controls. *Compared to controls and ^compared to LPS treatment alone, p < 0.05.

Article Snippet: Commercially available enzyme-linked immunosorbent assays were used according to the manufacturers’ protocols for measuring sCD14 (R&D System, Cat#DC140, Minneapolis, MN), and sCD163 (MyBioSource, Cat # MBS728094, San Diego, CA).

Techniques: Isolation

Journal: Scientific Reports

Article Title: Quantity of alcohol drinking positively correlates with serum levels of endotoxin and markers of monocyte activation

doi: 10.1038/s41598-017-04669-7

Figure Lengend Snippet: Relationship between serum LPS and sCD14 ( A ) and sCD163 ( B ). The relationship between serum LPS and sCD14/sCD163 was not linear; however, the correlation was evident when the serum LPS > 2 EU/ml which is consistent with the range we observed subjects with excessive drinking.

Article Snippet: Commercially available enzyme-linked immunosorbent assays were used according to the manufacturers’ protocols for measuring sCD14 (R&D System, Cat#DC140, Minneapolis, MN), and sCD163 (MyBioSource, Cat # MBS728094, San Diego, CA).

Techniques:

Journal: Scientific Reports

Article Title: Quantity of alcohol drinking positively correlates with serum levels of endotoxin and markers of monocyte activation

doi: 10.1038/s41598-017-04669-7

Figure Lengend Snippet: Relationship between serum LPS ( A ), sCD14 ( B ), and sCD163 ( C ) and quantity of alcohol consumption during the last 30 days as measured by timeline follow back. The serum levels of LPS, sCD14, and sCD163 were positively correlated with the quantity of alcohol consumption.

Article Snippet: Commercially available enzyme-linked immunosorbent assays were used according to the manufacturers’ protocols for measuring sCD14 (R&D System, Cat#DC140, Minneapolis, MN), and sCD163 (MyBioSource, Cat # MBS728094, San Diego, CA).

Techniques:

Journal: Scientific Reports

Article Title: Quantity of alcohol drinking positively correlates with serum levels of endotoxin and markers of monocyte activation

doi: 10.1038/s41598-017-04669-7

Figure Lengend Snippet: Serum levels of LPS, sCD14, and sCD163 in response to alcohol cessation. Abstinence was associated with significant decline in the levels of LPS ( A ), sCD14 ( B ) and sCd163 ( C ).

Article Snippet: Commercially available enzyme-linked immunosorbent assays were used according to the manufacturers’ protocols for measuring sCD14 (R&D System, Cat#DC140, Minneapolis, MN), and sCD163 (MyBioSource, Cat # MBS728094, San Diego, CA).

Techniques:

Journal: Experimental & molecular medicine

Article Title: TLR7-dependent eosinophil degranulation links psoriatic skin inflammation to small intestinal inflammatory changes in mice.

doi: 10.1038/s12276-024-01225-y

Figure Lengend Snippet: Fig. 1 Psoriatic skin inflammation induces changes in the intestinal microenvironment. a, b Serum concentrations of soluble CD14, calprotectin, and zonulin in healthy individuals and patients with psoriasis. c Schematic of imiquimod (IMQ)-induced psoriatic inflammation. d Serum fluorescein isothiocyanate (FITC)-dextran fluorescence and soluble CD14 and calprotectin concentrations in mice. e Images of hematoxylin and eosin-stained tissues from the small intestine (SI) and large intestine (LI). Scale bars, 124.5 μm. f Ratio of villus length to crypt length. Three to five villi and crypts were analyzed per section. g Calprotectin concentrations in stool. h Sequencing of the stool microbiota of the SI and LI of mice treated with IMQ (n = 3) or vehicle cream (n = 4). Image of the cytokine array membrane (i) and the quantified density (j). The data are presented as the means ± SDs. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 according to unpaired t tests (b, d, f, h, and CCL2 and CXCL9 in j), Mann–Whitney tests (calprotectin in b and j), or one-way ANOVA with Bonferroni’s multiple comparisons (g).

Article Snippet: A Quantikine Mouse CD14 ELISA Kit (R&D Systems, Minneapolis, MN, USA; cat# MC140) and a DuoSet Mouse Calprotectin ELISA Kit (R&D Systems; cat# DY8596-05) were used according to the manufacturer’s instructions.

Techniques: Staining, Sequencing, Cream, Membrane, MANN-WHITNEY