Journal: Clinical Medicine Insights. Arthritis and Musculoskeletal Disorders

Article Title: Macroscopical, Histological, and In Vitro Characterization of Nonosteoarthritic Versus Osteoarthritic Hip Joint Cartilage

doi: 10.4137/CMAMD.S29844

Figure Lengend Snippet: Expression of complement receptors (C5aR and C3aR) and CRPs (CD46, CD55, and CD59) in cartilage in situ. FNF and OA cartilage sections immunolabeled for ( A 1–2 ) C5aR, ( B 1–2 ) C3aR, ( C 1–2 ) CD46, ( D 1–2 ) CD55, and ( E 1–2 ) CD59. Notes: Cell nuclei were counterstained using DAPI (blue). Scale bar: 200 µm. Negative controls (B 3 ) performed by using only the secondary antibody revealed no unspecific staining.

Article Snippet: The primary antibodies (rabbit anti-human C3aR [1:30, Assay Biotechnology], mouse anti-human C5aR [1:100, GeneTex, Biozol], mouse anti-human CD46, CD59 [1:20 and 1:40, both AbD Serotec], and goat anti-human CD55 [1:40, R&D Systems]) were diluted in blocking buffer containing 0.1% Triton X-100 (Sigma-Aldrich) and incubated overnight at 4 °C in a humidified chamber.

Techniques: Expressing, In Situ, Immunolabeling, Staining

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: CD55 expression in human lung cancer tissue.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Expressing, Staining

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: CD55 expression in human lung cancer tissue. ( a ) Immunohistochemical analysis of CD55 in lung tissue. i, normal lung; ii, lung adenosquamous carcinoma; iii, lymph node metastatic carcinoma from lung adenocarcinoma; iv, lung mucoepidermoid carcinoma; v, lung squamous cell; vi, lung adenocarcinoma; vii, lung large cell carcinoma; viii, lung bronchioloalveolar carcinoma. Scale bars = 100 μm. Insets are magnified images demonstrating CD55 expression on the cell membrane and in the cytoplasm. ( b ) Immunohistochemical analysis of CD55 expression in normal organs. i, spleen; ii, lung; iii, skeletal muscle; iv, kidney; v, rectum; vi, colon; vii, stomach; viii, liver. Scale bars = 100 μm.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Expressing, Immunohistochemical staining, Membrane

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Development of chimeric CD55-specific monoclonal antibodies. ( a ) Phage ELISA results for individual clones tested against recombinant human CD55. A total of 384 clones were tested for binding to recombinant human CD55 (Supplementary Fig. ). The data for 36 clones are shown. ( b ) Reactivity of three anti-CD55 IgGs to recombinant human CD55 analyzed by antigen-binding ELISA. ( c ) SDS-PAGE analysis of the Ab1 anti-CD55 IgG antibody. ( d ) Representative flow cytometry analysis of H460 and H69 cells stained with the Ab1 anti-CD55 antibody.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Bioprocessing, Enzyme-linked Immunosorbent Assay, Clone Assay, Recombinant, Binding Assay, SDS Page, Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Characterization of 177 Lu-DTPA-anti-CD55 in vitro . ( a ) Representative results of three independent Lindmo assays to examine binding of 177 Lu-anti-CD55 to H460 cells. ( b ) Representative results of three independent saturation binding assays of 177 Lu-anti-CD55 in H460 cells. ( c ) Blocking assays in H460, H358, H69, and WI-38 cells (n = 3; **P < 0.01; Student’s t test). The results are presented as the mean ± standard error of the mean (SEM; error bars).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: In Vitro, Binding Assay, Blocking Assay

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Biodistribution analysis of 177 Lu-anti-CD55 in vivo . ( a ) The biodistribution of 177 Lu-anti-CD55 in tissue from pleural metastatic mice (n = 3–6 per condition). The results are presented as the mean ± SD (error bars). ( b ) The biodistribution of 177 Lu-anti-CD55 in normal lung and lung tumor tissue from pleural metastatic mice (n = 3–6 per condition; **P < 0.01; Student’s t test). The results are presented as the mean ± SD (error bars). ( c ) Excretion and residual radioactivity after injection of 177 Lu-anti-CD55 into mice. ( d ) Micro-SPECT/CT images of pleural metastatic or control mice treated with 177 Lu-anti-CD55. The amount of radioactivity is calculated in MBq/cc.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: In Vivo, Radioactivity, Injection, Micro-SPECT, Control

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Biodistribution of the 177 Lu-anti-CD55 antibody in a pleural metastatic mouse model.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques:

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Effects of the 177 Lu-anti-CD55 antibody on invasion and migration in lung cancer cells. ( a ) The invasive front of squamous cell carcinoma in the lung (arrows). Scale bars = 50 μm. ( b ) Invasion and migration of H460 cells treated with IgG, anti-CD55, or 177 Lu-anti-CD55. ( c ) Invasion and migration were quantified by counting cells in ten randomly selected regions (***P < 0.001; Student’s t test). The results are presented as the means ± SEM (error bars).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Migration

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Therapeutic efficacy of the 177 Lu-anti-CD55 antibody in vitro and in vivo . ( a ) Assays of H460 lung cancer cell viability after treatment with IgG, 177 Lu-IgG, anti-CD55, or 177 Lu-anti-CD55. α-CD55 (μg/ml) indicates the concentration of all the antibodies, IgG, 177 Lu-IgG, α-CD55, and 177 Lu-α-CD55. 177 Lu-α-CD55 (MBq) indicates the corresponding amounts of radioactivity of 177 Lu-α-CD55 for the indicated concentration (n = 3; **P < 0.01; Student’s t test). The results are presented as the mean ± SEM (error bars). ( b ) Assays of H358 cell viability after treatment with IgG, 177 Lu-IgG, anti-CD55, or 177 Lu-anti-CD55 (n = 3; *P < 0.05; Student’s t test). The results are presented as the mean ± SEM (error bars). ( c ) Effects of 177 Lu-anti-CD55 on the survival of H460 bearing-early pleural metastatic mice (n = 10 for each group; ***P < 0.001; Log-rank (Mantel-Cox) test). ( d ) Effects of 177 Lu-anti-CD55 on the survival of pleural metastatic mice (n = 10 for each group; **P < 0.01; Log-rank (Mantel-Cox) test).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Drug discovery, In Vitro, In Vivo, Concentration Assay, Radioactivity

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Combinatorial effects of 177 Lu-anti-CD55 and cisplatin in vitro and in vivo . ( a ) Cell viability assays of H460 lung cancer cells treated with 0.1 MBq of 177 Lu-anti-CD55 in the presence or absence of 3 μM cisplatin (n = 3; *P < 0.05; Student’s t test). The results are presented as the mean ± SEM (error bars). ( b ) Cell viability assays of H358 lung cancer cells treated with 0.1 MBq of 177 Lu-anti-CD55 in the presence or absence of 3 μM cisplatin (n = 3; *P < 0.05; Student’s t test). The results are presented as the mean ± SEM (error bars). ( c ) Effects of 177 Lu-anti-CD55 in the presence or absence of cisplatin on the survival of early pleural metastatic mice (n = 10 for each group; ***P < 0.001; Log-rank (Mantel-Cox) test). ( d ) Analysis of the survival of pleural metastatic mice after treatment with 177 Lu-anti-CD55 in the presence or absence of cisplatin (n = 10 for each group).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: In Vitro, In Vivo

Journal: The Journal of Experimental Medicine

Article Title: Loss of decay-accelerating factor triggers podocyte injury and glomerulosclerosis

doi: 10.1084/jem.20191699

Figure Lengend Snippet: ADR induces PLAD-dependent cleavage of DAF. (A and B) Representative images (A) and distribution of DAF expression (B) quantified in hiPod exposed to vehicle, ADR (0.3 µg/ml), PLADi (1 µM), or ADR + PLADi (0.3 µg/ml in 1 µM) for 24 h. DAF IF signal was normalized to actin expression pixel by pixel, and the MFI for each cell was computed. Results are representative of two independent experiments with similar results. (C and D) Representative blots (C) and densitometric analysis (D) of PLAD expression in hiPod cell lysates previously exposed to vehicle or ADR for 24 h. (E and F) Representative blots (E) and densitometric analysis (F) of DAF in the supernatants of hiPod exposed to ADR for 24 h with or without PLADi (WB). (G) Representative blot of DAF in the urine from BALB/c male mice at 2 wk after treatment with vehicle or ADR compared with recombinant mouse DAF (rDAF). In each group, we pooled and concentrated urine samples from eight mice (see Materials and methods). All experimental data were verified in at least three independent experiments. *P < 0.05; n.s., not significant. Scale bars: 50 µm. Error bars are SEM.

Article Snippet: The urine from mouse and human samples was separated on 4–20% precast Protean TGX gels (Bio-Rad Laboratories) with recombinant mouse (55 μg/ml, 5490-CD-050; R&D Systems) or human (15 μg/ml, SRP6437; Sigma-Aldrich) CD55 protein, respectively.

Techniques: Expressing, Recombinant

Journal: The Journal of Experimental Medicine

Article Title: Loss of decay-accelerating factor triggers podocyte injury and glomerulosclerosis

doi: 10.1084/jem.20191699

Figure Lengend Snippet: FSGS in humans is associated with DAF down-regulation and complement activation. (A–D) C3 (A), C3aR (B), C5aR (C), and DAF mRNA (D) expression in glomeruli of human biopsy specimens with pathological diagnosis of FSGS or diabetic kidney disease compared with normal kidneys. Data are from previously published microarray studies by and were subjected to further analysis using Nephroseq. (E–H) Representative renal staining and data quantification for C3d (IF; E and F) and DAF (immunohistochemistry; G and H) in patients with FSGS ( n = 18) and in kidneys from healthy renal donors ( n = 10). (I) Correlation between protein and C3a in urine samples from 27 patients with FSGS taken at the time of kidney biopsy (before therapy). (J and K) Differences in proteinuria (J) and urinary C3a (K) measured before versus 3–6 mo after steroid therapy in a subset of 13 patients with FSGS. (L) Correlation between the change in proteinuria and change in urinary C3a before and after therapy for each of the same 13 patients. (M) Representative blot of DAF in the urine from healthy control individuals and patients with FSGS compared with recombinant human DAF (rDAF). In each group, we pooled and concentrated urine samples from five and five subjects, respectively (see Materials and methods). *P ≤ 0.05. Scale bars: 25 μm. Error bars are SEM.

Article Snippet: The urine from mouse and human samples was separated on 4–20% precast Protean TGX gels (Bio-Rad Laboratories) with recombinant mouse (55 μg/ml, 5490-CD-050; R&D Systems) or human (15 μg/ml, SRP6437; Sigma-Aldrich) CD55 protein, respectively.

Techniques: Activation Assay, Expressing, Biomarker Discovery, Microarray, Staining, Immunohistochemistry, Control, Recombinant

Journal: bioRxiv

Article Title: Actinotrichia-independent developmental mechanisms of spiny rays facilitate the morphological diversification of Acanthomorpha fish fins

doi: 10.1101/2025.03.01.640274

Figure Lengend Snippet: a Distribution of the actinotrichia at the tips of spiny rays (left) and soft rays (right) at three stages of fin deveolpent in fixed samples; the lower pair of panels of each stage are magnified images of the yellow dashed box in the upper panels. Spiny-ray bones are outlined in orange. b, c Actinotrichia distribution and spiny- and soft-ray bone morphology in wild-type fish (panel b , n = 8) and in the actinodin1 −/− / actinodin2 +/- knockout fish (panel c , n = 6). Actinotrichia were labeled with DAFFM DA (green), and the spiny- and soft-ray bones were labeled with alizarin red (magenta). d Schematic illustration of the spiny- and soft-ray morphology in fish with a loss of actinotrichia.

Article Snippet: To observe the actinotrichia, vital staining with 5 μM of DAFFM DA (Goryo Chemical, #SK1003-01) was used.

Techniques: Knock-Out, Labeling

Journal: bioRxiv

Article Title: Actinotrichia-independent developmental mechanisms of spiny rays facilitate the morphological diversification of Acanthomorpha fish fins

doi: 10.1101/2025.03.01.640274

Figure Lengend Snippet: Sections of the second dorsal fin showing the structural organization of the soft-ray tip in larvae at DF-st4. The same TEM images are presented corresponding to the lower panels, with the soft ray (light blue), actinotrichia (magenta), and basement membrane (yellow) highlighted in different colors. The numbers of each panels (#1, #2, #3) in correspond to the numbers in yellow dashed boxes. Mes, mesenchymal.

Article Snippet: To observe the actinotrichia, vital staining with 5 μM of DAFFM DA (Goryo Chemical, #SK1003-01) was used.

Techniques: Membrane

Journal: bioRxiv

Article Title: Actinotrichia-independent developmental mechanisms of spiny rays facilitate the morphological diversification of Acanthomorpha fish fins

doi: 10.1101/2025.03.01.640274

Figure Lengend Snippet: In spiny-ray development, mesenchymal cells (green) condense at the tips of spiny-ray bones, and the layer of extracellular matrix (ECM) (yellow) surrounds mesenchymal cells. In soft-ray development, the actinotrichia (magenta) are distributed at the tips of soft-ray bones and serve as scaffolds for mesenchymal cells (green). Micro-CT scan image were used to schematically illustrate the fin bones.

Article Snippet: To observe the actinotrichia, vital staining with 5 μM of DAFFM DA (Goryo Chemical, #SK1003-01) was used.

Techniques: Micro-CT

Journal: bioRxiv

Article Title: Actinotrichia-independent developmental mechanisms of spiny rays facilitate the morphological diversification of Acanthomorpha fish fins

doi: 10.1101/2025.03.01.640274

Figure Lengend Snippet: a Schematic illustration of the obtaining the actinodin1 ( and1 ) and actinodin2 ( and2 ) knockout fish. b Actinotrichia distribution and spiny- and soft-ray bone morphology in the and1 −/− / and2 +/+ knockout fish. Actinotrichia were labeled with DAFFM DA (green), and the spiny- and soft-ray bones were labeled with alizarin red (magenta). White brackets and arrows indicate actinotrichia. c Sequences of actinodin1 −/− / actinodin2 +/+ knockout fish, showing a mutation spectrum revealed by Sanger sequencing. Sequences in blue and green indicate the PAM and the recognition sequence for the sgRNA target, respectively. a.a., amino acids.

Article Snippet: To observe the actinotrichia, vital staining with 5 μM of DAFFM DA (Goryo Chemical, #SK1003-01) was used.

Techniques: Knock-Out, Labeling, Mutagenesis, Sequencing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 4. CML triggered by anti–EGFR-IgG3 negatively correlates with CD55 and CD59 expression levels. (A) Surface expression levels of CD46, CD55, and CD59 on analyzed cell lines were quantified by calibrated flow cytometry. Means 6 SEM of at least three independent experiments are presented. (B) Correlations between CD46, CD55, or CD59 and anti–EGFR-IgG3–mediated CDC were calculated for all four cell lines. CDC results at 2 mg/ml Ab concentration were taken from experiments presented in Fig. 2. (C) A431 cells were seeded into 10-cm plates and grown overnight. On the following day, cells were transfected with 50 nM control siRNA or with single siRNAs specific for CD46, CD55, CD59, or with a combination of all three mCRP-specific siRNAs for 72 h. Efficiency of siRNA-induced knockdown was analyzed by direct flow cytometry using fluorochrome-labeled, mCRP- specific Abs (CD46-Pacific blue, CD55-PE, CD59-FITC), or respective control Abs. (D–G) CDC against control siRNA or mCRP-specific, siRNA- transfected A431 cells was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human serum and anti–EGFR-IgG1 (upper panels), anti– EGFR-IgG3 (lower panels), as well as the respective control Abs at increasing concentrations. (H) Concentration-dependent binding of CD55-Ab (BRIC216, mouse IgG1) to A431 cells was analyzed by indirect immunofluorescence. Results from one representative experiment are presented. (I) CDC triggered by anti–EGFR-IgG1, anti–EGFR-IgG3, or respective control Abs (all at 66.67 nM) against A431 cells in the presence of saturating concentrations of CD55-Ab or a control Ab (both at 66.67 nM) was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human (Figure legend continues)

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Expressing, Cytometry, Concentration Assay, Transfection, Control, Knockdown, Labeling, Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 5. CD55 dampens anti–EGFR-IgG3–triggered CML and promotes C1q-dependent induction of AP amplification. BHK-EGFR+ #5 cells were transiently transfected with a control vector or a CD55 vector for 48 h. (A) Cell-surface expression of CD55 was analyzed by direct flow cytometry using PE-conjugated CD55-specific or control Abs. (B–D) The influence of CD55 overexpression on anti–EGFR-IgG3–mediated CDC was investigated by [51Cr] release assays either (B) in an Ab concentration–response curve, (C) in a time-dependent manner, or (D) in serum titration experiments. (E–H) The influence of the alternative complement pathway inhibitor CRIg (E and G), the presence of C1q in serum (F; at 66.67 nM Ab concentration; mean 6 SEM of triplicates), as well as of factor B (H; 13.33 nM Ab concentration, 12.5% v/v factor B–depleted serum, 200 mg/ml factor B), on anti–EGFR-IgG3–mediated CDC was analyzed using either (E and F) control vector–transfected or CD55 vector–transfected BHK-EGFR+ #5 cells or (G and H) DiFi cells (66.67 nM Ab concentration). (I) Deposition of factor Bb on control vector– or CD55 vector–transfected BHK-EGFR+ #5 cells was analyzed by flow cytometry. Relative deposition levels were calculated by equating RFI measured in the absence of Ab with 100%. Results are presented as mean 6 SEM of at least three independent experiments with different blood donors. *p # 0.05 anti–EGFR-IgG3 versus respective control Ab; (B–D, I) #p # 0.05 control vector versus CD55 vector; (E and G) #p # 0.05 without CRIg-Fc versus CRIg-Fc; (H) #p # 0.05 w/o factor B versus with factor B.

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Transfection, Control, Plasmid Preparation, Expressing, Cytometry, Over Expression, Concentration Assay, Titration

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 6. Overview of complement activation by human anti–EGFR-IgG3 in the context of CD55 expression. On CD55-deficient target cells (left panel), anti–EGFR-IgG3 mediates strong C3b but low C4b deposition and induces assembly of classical and alternative C3 convertases, predominantly resulting in the induction of fast and efficient CDC via the classical pathway of complement activation. In contrast, on CD55-expressing target cells (right panel), CD55 mainly accelerates the decay of low amounts of classical C3 convertases, leading to amplification of the AP and finally to slow and inefficient CDC induction.

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Activation Assay, Expressing