Journal: International journal of molecular sciences

Article Title: Peritoneal Expression of Membrane Complement Regulators Is Decreased in Peritoneal Dialysis Patients with Infected Peritonitis.

doi: 10.3390/ijms24119146

Figure Lengend Snippet: Figure 1. Correlations of expressions of membrane complement regulators (CRegs), CD46, CD55, and CD59, deposition of complement (C) activation products such as aC3 and C5b-9, and levels of sC5b-9 in peritoneal dialysis fluid (PDF) with tissue injury score and comparison of sC5-9 levels in PDF between Groups P and NP. Expressions of each of the CRegs CD46, CD55, and CD59 showed good inverse correlations with the severity score for peritoneal injuries ((A–C) respectively). In contrast, deposition of C activation products such as activated C3 (aC3) and C5b-9 correlated with the severity score of peritoneal tissue injures ((D,E) respectively). Adjusted PDF levels of sC5b-9 (sC5b-9/TP) also correlated with the severity score for peritoneal injures (F). Between Groups P and NP, the tissue injury score and PDF levels of sC5b-9/TP were significantly different ((G,H), respectively). Dots in the graph show outliers. Outliers were defined as values greater than the 75th percentile plus 1.5 times the interquartile range. #, p < 0.05.

Article Snippet: Monoclonal Ab (mAb) mouse antihuman CD46 (MEM 258) was purchased from BIO-RAD (Hercules, CA, USA).

Techniques: Membrane, Activation Assay, Comparison

Journal: International journal of molecular sciences

Article Title: Peritoneal Expression of Membrane Complement Regulators Is Decreased in Peritoneal Dialysis Patients with Infected Peritonitis.

doi: 10.3390/ijms24119146

Figure Lengend Snippet: Figure 4. Expressions of CRegs CD46, CD55, and CD59 in those with and without peritonitis and depositions of complement (C) activation products. Frames (A–E) and (F–J) represent peritoneal tissues in Group P and Group NP, respectively. Frames (A,F), frames (B,G), frames (C,H), frames (D,I), and frames (E,J) show expressions of CD46, CD55, and CD59 and depositions of activated C3 (aC3) and C5b- 9, respectively. Graphs (K–O) show results for expressions of CD46, CD55, and CD59 and deposition of aC3 and C5b-9, respectively. Original magnification of (A–C) and (F–H) is ×400, while that of (D,E,I,J) is ×100. Inserted frames in right upper corners of frames (F–H) are at double the original magnification. Scale bars of 50 µm and 200 µm are shown in the left upper corner of frames (A,D), respectively. * shows peritoneal cavity side. Dots in the graph show outliers. Outliers were defined as values greater than the 75th percentile plus 1.5 times the interquartile range. # p < 0.05.

Article Snippet: Monoclonal Ab (mAb) mouse antihuman CD46 (MEM 258) was purchased from BIO-RAD (Hercules, CA, USA).

Techniques: Activation Assay

Journal: International journal of molecular sciences

Article Title: Peritoneal Expression of Membrane Complement Regulators Is Decreased in Peritoneal Dialysis Patients with Infected Peritonitis.

doi: 10.3390/ijms24119146

Figure Lengend Snippet: Figure 5. Pathological changes with accumulation of inflammatory cells of peritonitis and expression of membrane complement regulators (CRegs) caused by Candida spp. or P. aeruginosa (Group P1) and Gram-positive cocci (Group P2). Graphs (A,B) show the tissue injury scores and the proportion of cytokeratin-positive mesothelial cells in the peritoneum, respectively. Graph (C–E) shows the numbers of esterase-positive neutrophils, CD3-positive pan-T cells, and CD68-positive macrophages, respectively. Graphs (F,G) show the scores of fibrin deposition and fibrosis in the peritoneum, respectively. Graphs (H–J) show CRegs CD46, CD55, and CD59, respectively. Group P1 shows cases with fungal infection or P. aeruginosa and Group P2 shows cases with Gram-positive cocci. Dots in the graph show outliers. Outliers were defined as values greater than the 75th percentile plus 1.5 times the interquartile range. #, p < 0.05.

Article Snippet: Monoclonal Ab (mAb) mouse antihuman CD46 (MEM 258) was purchased from BIO-RAD (Hercules, CA, USA).

Techniques: Expressing, Membrane, Infection

Journal: International Journal of Molecular Sciences

Article Title: Peritoneal Expression of Membrane Complement Regulators Is Decreased in Peritoneal Dialysis Patients with Infected Peritonitis

doi: 10.3390/ijms24119146

Figure Lengend Snippet: Correlations of expressions of membrane complement regulators (CRegs), CD46, CD55, and CD59, deposition of complement (C) activation products such as aC3 and C5b-9, and levels of sC5b-9 in peritoneal dialysis fluid (PDF) with tissue injury score and comparison of sC5-9 levels in PDF between Groups P and NP. Expressions of each of the CRegs CD46, CD55, and CD59 showed good inverse correlations with the severity score for peritoneal injuries (( A – C ) respectively). In contrast, deposition of C activation products such as activated C3 (aC3) and C5b-9 correlated with the severity score of peritoneal tissue injures (( D , E ) respectively). Adjusted PDF levels of sC5b-9 (sC5b-9/TP) also correlated with the severity score for peritoneal injures ( F ). Between Groups P and NP, the tissue injury score and PDF levels of sC5b-9/TP were significantly different (( G , H ), respectively). Dots in the graph show outliers. Outliers were defined as values greater than the 75th percentile plus 1.5 times the interquartile range. # , p < 0.05.

Article Snippet: Monoclonal Ab (mAb) mouse anti-human CD46 (MEM 258) was purchased from BIO-RAD (Hercules, CA, USA).

Techniques: Membrane, Activation Assay, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Peritoneal Expression of Membrane Complement Regulators Is Decreased in Peritoneal Dialysis Patients with Infected Peritonitis

doi: 10.3390/ijms24119146

Figure Lengend Snippet: Expressions of CRegs CD46, CD55, and CD59 in those with and without peritonitis and depositions of complement (C) activation products. Frames ( A – E ) and ( F – J ) represent peritoneal tissues in Group P and Group NP, respectively. Frames ( A , F ), frames ( B , G ), frames ( C , H ), frames ( D , I ), and frames ( E , J ) show expressions of CD46, CD55, and CD59 and depositions of activated C3 (aC3) and C5b-9, respectively. Graphs ( K – O ) show results for expressions of CD46, CD55, and CD59 and deposition of aC3 and C5b-9, respectively. Original magnification of ( A – C ) and ( F – H ) is ×400, while that of ( D , E , I , J ) is ×100. Inserted frames in right upper corners of frames ( F – H ) are at double the original magnification. Scale bars of 50 μm and 200 μm are shown in the left upper corner of frames ( A , D ), respectively. * shows peritoneal cavity side. Dots in the graph show outliers. Outliers were defined as values greater than the 75th percentile plus 1.5 times the interquartile range. # p < 0.05.

Article Snippet: Monoclonal Ab (mAb) mouse anti-human CD46 (MEM 258) was purchased from BIO-RAD (Hercules, CA, USA).

Techniques: Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Peritoneal Expression of Membrane Complement Regulators Is Decreased in Peritoneal Dialysis Patients with Infected Peritonitis

doi: 10.3390/ijms24119146

Figure Lengend Snippet: Pathological changes with accumulation of inflammatory cells of peritonitis and expression of membrane complement regulators (CRegs) caused by Candida spp. or P. aeruginosa (Group P1) and Gram-positive cocci (Group P2). Graphs ( A , B ) show the tissue injury scores and the proportion of cytokeratin-positive mesothelial cells in the peritoneum, respectively. Graph ( C – E ) shows the numbers of esterase-positive neutrophils, CD3-positive pan-T cells, and CD68-positive macrophages, respectively. Graphs ( F , G ) show the scores of fibrin deposition and fibrosis in the peritoneum, respectively. Graphs ( H – J ) show CRegs CD46, CD55, and CD59, respectively. Group P1 shows cases with fungal infection or P. aeruginosa and Group P2 shows cases with Gram-positive cocci. Dots in the graph show outliers. Outliers were defined as values greater than the 75th percentile plus 1.5 times the interquartile range. # , p < 0.05.

Article Snippet: Monoclonal Ab (mAb) mouse anti-human CD46 (MEM 258) was purchased from BIO-RAD (Hercules, CA, USA).

Techniques: Expressing, Membrane, Infection

Journal: Cancer Immunology Research

Article Title: MicroRNAs Affect Complement Regulator Expression and Mitochondrial Activity to Modulate Cell Resistance to Complement-Dependent Cytotoxicity

doi: 10.1158/2326-6066.cir-18-0818

Figure Lengend Snippet: Figure 6. Modulation of the membrane complement regulatory proteins by microRNAs. The expression of CD46 (A), CD55 (B), and CD59 (C) in lysates of K562 cells transfected with microRNA inhibitors or with control, determined by the proteomic analysis described in Materials and Methods, is shown as the mean SD of log2 of intensity value units of five independent experiments. , P < 0.05; , P < 0.01, relative to control (Student t test). NS, not significant. D, K562 cells were transfected with miR-616 inhibitor plasmid or with control (C) plasmid as a negative control. After 48 hours, the cells were labeled with anti-CD46, anti-CD55, or anti-CD59 and then with fluorescently labeled secondary antibody. Cells were then analyzed by flow cytometry, and the mean fluorescence intensity (MFI) values, representative of three independent experiments, were determined. The expression of each regulator in anti–miR-616–treated cells was normalized to their levels in control cells (set as 100). , P < 0.05; , P < 0.01, relative to control (Student t test). E–G, K562 cells were transfected with a miR-150 expression plasmid or a control plasmid. After 24 hours, the cells were labeled with mouse anti-CD46 (E), anti-CD55 (F), or anti-CD59 (G) and fluorescently labeled secondary antibody. Cells were then analyzed by flow cytometry, and MFI values, representative of three independent experiments, were determined. , P < 0.05, relative to control (Student t test).

Article Snippet: Mouse monoclonal antihuman CD46 (clone MEM-258), anti-human CD55 (clone 67), and anti-human CD59 (clone MEM-43) antibodies were purchased from AbD Serotec.

Techniques: Membrane, Expressing, Transfection, Control, Plasmid Preparation, Negative Control, Labeling, Cytometry