Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Virulence of S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) in mice a

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Mutagenesis

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Opsonophagocytosis of fully encapsulated type 3 pneumococci and unencapsulated and PspA-deficient mutants in NHS. Opsonization, determined by flow cytometry, is expressed as opsonic index (see Materials and Methods for details). Bars represent means ± standard errors of three separate experiments. S3 (ATCC 6303) and WU2 are fully encapsulated parent strains (Caps+/PspA+), JY1123 is a PspA-deficient mutant of WU2 (Caps+/PspA−), and DW3.8 is an unencapsulated mutant of WU2 (Caps−/PspA−). JY1123 and DW3.8 significantly different from WU2 (∗∗∗, P < 0.002; ∗∗∗∗, P < 0.001 [Student’s t test]).

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Flow Cytometry, Mutagenesis

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Activation Assay, Mutagenesis

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Formation of TCC by the type 3 pneumococcal strains S3 (ATCC 6303) and WU2 (both Caps+/PspA+), the unencapsulated mutant DW3.8 (Caps−/PspA−) of WU2, and the PspA-deficient mutant JY1123 (Caps+/PspA−) of WU2. TCC were determined by ELISA after incubation of pneumococci with NHS for 10 min at 37°C, and TCC formation was expressed as the ELISA titer (means ± standard errors of three independent experiments [horizontal top axis]). Control, no pneumococci added; Zymosan, positive control for TCC formation. DW3.8 was significantly different from ATCC 6303, WU2, and JY1123 (∗ P < 0.05 [Student’s t test]).

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Mutagenesis, Enzyme-linked Immunosorbent Assay, Incubation, Positive Control

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Effect of trypsin treatment of pneumococci on complement activation by type 3 strains ATCC 6303 and WU2 (both Caps+/PspA+) and unencapsulated and PspA-deficient mutants of WU2. Complement activation, determined by the hemolytic assay, is expressed as a Z value, representing the number of active hemolytic sites per erythrocyte (y axis). Bars represent mean is ± standard errors of three independent experiments. S3 (ATCC 6303) and WU2 are fully encapsulated parent strains, JY1123 is a PspA-deficient mutant of WU2 (Caps+/PspA−), and DW3.8 is an unencapsulated mutant of WU2 (Caps−/PspA−). Trypsin-treated bacteria (open bars) were significantly different from untreated bacteria (hatched bars), and both buffer-treated JY1123 and DW3.8 were significantly different from buffer-treated WU2 (∗∗, P < 0.01: ∗∗∗∗, P < 0.001 [Student’s t test]).

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Activation Assay, Hemolytic Assay, Mutagenesis

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Capsular polysaccharide content of S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) before and after trypsin treatment

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Mutagenesis

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Effect of trypsin treatment on opsonophagocytosis of type 3 strain S3 (ATCC 6303) in NHS. Opsonization, determined by flow cytometry, is expressed as the opsonic index (see Materials and Methods for details). Bars represent means ± standard errors of three separate experiments. S3/VSB++, S3 (ATCC 6303) pretreated with buffer; S3/trypsin, S3 (ATCC 6303) pretreated with trypsin. Trypsin-treated bacterial were significantly different from untreated bacteria (∗∗, P < 0.01 [Student’s t test]).

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Flow Cytometry

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Effect of trypsin treatment on the binding of complement-regulatory protein factor H by type 3 pneumococcal strains S3 (ATCC 6303) and WU2 (both Caps+/PspA+), the unencapsulated mutant DW3.8 (Caps−/PspA−) of WU2 and the PspA-deficient mutant JY1123 (Caps+/PspA−) of WU2. Factor H binding was determined by flow cytometry and is expressed as fluorescence units (means ± standard errors of three independent experiments) on the y axis. DW3.8 was significantly different from S3 (ATCC 6303), WU2, and JY1123 (∗∗, P < 0.01 [Student’s t test]). Trypsin-treated bacteria were significantly different from buffer-treated bacteria (∗, P < 0.05; ∗∗∗∗, P < 0.001 [Student’s t test]).

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Binding Assay, Mutagenesis, Flow Cytometry, Fluorescence

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Effect of trypsin treatment on the PspA content of pneumococci. The PspA content of pneumococcal strains ATCC 6303 (S3) (Caps+/PspA+), WU2 (Caps+/PspA+), JY1123 (Caps+/PspA+), and DW3.8 (Caps+/PspA+) was determined by FACS analysis with monoclonal antibody XiR 278 (see Materials and Methods). Results are expressed as arbitrary fluorescence units. Bars denote means ± standard errors of three independent experiments. Trypsin-treated S3 and WU2 were significantly different from bacteria treated with buffer, and JY1123 and DW3.8 were significantly different from buffer-treated WU2 (∗∗∗∗, P < 0.001 [Student’s t test]).

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Fluorescence

Journal:

Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

doi:

Figure Lengend Snippet: Immunoblotting of cell wall proteins of type 3 pneumococcal strain ATCC 6303 with anti-human factor H antibody. Pneumococcal proteins blotted onto a nitrocellulose membrane were sequentially incubated with NHS, rabbit anti-human factor H antibody, and peroxidase-labeled goat anti-rabbit antibody. Left lane, high-range molecular mass markers (47 to 208 kDa); middle lane, pneumococcal proteins reactive with factor H (88, 150, and 196 kDa); right lane, low-range molecular mass markers (34.6 to 104 kDa). The molecular masses of low- and high-range marker proteins are indicated on right and left of the immunoblot, respectively.

Article Snippet: Complement activation induced by S. pneumoniae type 3 (ATCC 6303 and WU2) and type 3 mutant strains (JY1123 and DW3.8) a The role of the classic and alternative pathways in complement activation by the different pneumococcal strains was investigated by using EGTA- and EDTA-containing buffers during the activation step.

Techniques: Western Blot, Incubation, Labeling, Marker