Journal: bioRxiv

Article Title: Plasmodium falciparum exploits CD44 as a co-receptor for erythrocyte invasion

doi: 10.1101/2023.04.12.536503

Figure Lengend Snippet: (A) Mass spectrometry results from affinity purification experiments in which bead-immobilized rCD44-Fc or rFc were incubated with lysate from P. falciparum strain 3D7 schizont-stage parasites. Heat map shows the normalized spectral counts (based on protein size) of the most abundant P. falciparum proteins detected and their enrichment in rCD44-Fc compared to rFc. NaN indicates no counts. The table shows Log probability, raw number of spectra and unique peptides, and coverage of EBA-140 and EBA-175 proteins found in rCD44-Fc lane. (B) Western blot of independent affinity purification experiment in which bead-immobilized rCD44-Fc or rFc were incubated with P. falciparum schizont-stage lysate, followed by immunoblotting for EBA-175 or EBA-140. SN, supernatant. (C) Flow cytometry-based binding assays in which recombinant region RII of EBA-175-His (left) or EBA-140-His (right) were incubated with beads coated with rCD44-Fc, rFc, or no protein. Binding was detected using an anti-His antibody and a fluorescent secondary antibody. (D) Flow cytometry-based binding assays between rCD55-Fc or rFc and region RII of EBA-175-His (left) or EBA-140-His (right). (E) Binding assays of RII EBA-175 (left) or RII EBA-140 (right) with rCD44-Fc in the presence of anti-CD44 monoclonal antibody BRIC 222 or isotype control BRIC 170.

Article Snippet: Protein A Dynabeads (1.5 mg) were incubated with 8 µg rCD44-Fc (R&D systems 3660-CD), rCD44-Fc (SinoBiological 12211-H02H), rCD55-Fc (SinoBiological 10101-H02H), IgG 1 -Fc (R&D systems 110-HG), or no protein in 200 µl antibody binding and washing buffer (Invitrogen kit 10006D) for one hour at RT.

Techniques: Mass Spectrometry, Affinity Purification, Incubation, Western Blot, Flow Cytometry, Binding Assay, Recombinant, Protein Binding, Control

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: 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: Nature Communications

Article Title: IL-21R-STAT3 signalling initiates a differentiation program in uterine tissue-resident NK cells to support pregnancy

doi: 10.1038/s41467-023-42990-0

Figure Lengend Snippet: a RNA velocity of the differentiation trajectory of uterine trNK. b Pseudotime inference of uterine trNK differentiation in a two-dimensional state-space defined by Monocle2 (left), and ordering of each uterine trNK subgroup along the trajectory (right). c Distribution of each uterine trNK subgroup along the pseudotime trajectory. d Gene expression dynamics of uterine trNK along pseudotime trajectory. e Expression of the indicated effector genes in uterine trNK, respected to pseudotime coordinate. f Expression of the indicated transcription factors in uterine trNK, respected to pseudotime coordinate. g Expression of the indicated surface molecules in uterine trNK, respected to pseudotime coordinate. h Flow cytometry showing co-expression of 4-1BB and CD55 on uterine trNK in virgin and gd8.5 mice. i Giemsa-Wright staining showing the morphology of uterine trNK-V, trNK-D and trNK-T subgroups from gd8.5 dams. j Deconvolution of the transcriptomes (bulk RNA-Seq) of uterine trNK-V, trNK-D and trNK-T subgroups, using the gene signatures of virginal, differentiating, and terminal trNK subgroups (based on scRNA-Seq). Numbers indicate the percentages in each box. Data are shown as the mean ± SEM. Data are representative of two independent experiments ( a – j ) or at least three independent experiments ( h – i ). Source data are provided as a Source Data file.

Article Snippet: Antibodies specific to mouse CD45.2 (104), CD19 (eBio1D3), CD11b (M1/70), CD11c (N418), CD5 (53-7.3), Gr-1 (RB6-8C5), NKp46 (29A1.4), NK1.1 (PK136), 4-1BB (17B5), EOMES (Dan11mag), Ki-67 (SolA15) were purchased from eBioscience; antibodies specific to mouse CD3 (145-2C11), F4/80 (QA17A29), I-A/I-E (M5/114.15.2), CD49a (HMα1), IL-21R (4A9), CXCR6 (SA051D1), CD16/32 (93) were purchased from BioLegend; antibodies specific to mouse phospho-STAT3 (Tyr705) (4/P-STAT3) were purchased from BD Biosciences; antibody specific to mouse CD55 (076) was purchased from Sino Biological.

Techniques: Expressing, Flow Cytometry, Wright Stain, RNA Sequencing Assay

Journal: Nature Communications

Article Title: IL-21R-STAT3 signalling initiates a differentiation program in uterine tissue-resident NK cells to support pregnancy

doi: 10.1038/s41467-023-42990-0

Figure Lengend Snippet: a Flow cytometry and the corresponding statistics showing compositional change of uterine NK in virgin mice and pregnant mice at the indicated gestational days ( n = 3, 4, 4, and 3 per group). b Schematics of prediction for potential transcription regulators (related to Supplementary Fig. ). c Scatter plots showing the motif enrichment and expression of potential trNK-specific transcription regulators. d Dot plot showing the expression of cytokine receptors associated with STAT3 activation. e Flow cytometric histogram showing IL-21R expression on uterine trNK-V, trNK-D, trNK-T subgroups and cNK from gd8.5 mice ( n = 4 per group; * P = 0.0152, * P = 0.0483, ** P = 0.0011). f Quantitative PCR assessing Il21 expression in uteri at the indicated gestational days ( n = 5, 5, 3, 3, and 3 per group; * P = 0.0258, * P = 0.0377). g Flow cytometric and statistical analysis of STAT3 phosphorylation in uterine trNK. Uterine trNK are sorted from virgin and gd6.5 mice, and stimulated in presence or absence of recombinant IL-21 for 30 min ( n = 4 per group; * P = 0.0348, * P = 0.0316). ( h - k ) Uterine trNK-V subgroup is sorted from gd6.5 mice, and cultured in presence or absence of IL-21 and STAT3 inhibitor (iSTAT3) for 72 h. h Flow cytometric and statistical analysis showing the percentage of 4-1BB - CD55 - , 4-1BB + CD55 - , 4-1BB - CD55 + and 4-1BB + CD55 + trNK subgroups ( n = 4 per group; *** P = 0.0010, ** P = 0.0019, ** P = 0.0011, ** P = 0.0021, ** P = 0.0078, * P = 0.0122). i Dot plot depicting the gene expression changes of in vitro cultured uterine trNK ( h ). j Heatmap showing expression change of the STAT3-dependent and -independent genes ( i ) k Scoring of the STAT3-dependent IL-21-upregulated genes in gd8.5 uterine trNK. Numbers indicate the percentages in each box. Data are shown as the mean ± SEM. P -values are calculated by two-sided unpaired t -test, * P < 0.05, ** P < 0.01, *** P < 0.001. Data are representative of two independent experiments ( b – d , i – k ) or at least three independent experiments ( a , e – h ). Source data are provided as a Source Data file.

Article Snippet: Antibodies specific to mouse CD45.2 (104), CD19 (eBio1D3), CD11b (M1/70), CD11c (N418), CD5 (53-7.3), Gr-1 (RB6-8C5), NKp46 (29A1.4), NK1.1 (PK136), 4-1BB (17B5), EOMES (Dan11mag), Ki-67 (SolA15) were purchased from eBioscience; antibodies specific to mouse CD3 (145-2C11), F4/80 (QA17A29), I-A/I-E (M5/114.15.2), CD49a (HMα1), IL-21R (4A9), CXCR6 (SA051D1), CD16/32 (93) were purchased from BioLegend; antibodies specific to mouse phospho-STAT3 (Tyr705) (4/P-STAT3) were purchased from BD Biosciences; antibody specific to mouse CD55 (076) was purchased from Sino Biological.

Techniques: Flow Cytometry, Expressing, Activation Assay, Real-time Polymerase Chain Reaction, Recombinant, Cell Culture, In Vitro

Journal: Cell Reports Methods

Article Title: Sequence enrichment profiles enable target-agnostic antibody generation for a broad range of antigens

doi: 10.1016/j.crmeth.2023.100475

Figure Lengend Snippet:

Article Snippet: CD55 , Sino Biological , Cat# 10101-H08H.

Techniques: Produced, Recombinant, Sequencing, Software