Journal:

Article Title: Beneficial Immunomodulation by Streptococcus mutans anti-P1 Monoclonal Antibodies is Fc-independent and Correlates with Increased Exposure of a Relevant Target Epitope

doi: 10.4049/jimmunol.0803300

Figure Lengend Snippet: Host genetic background affects beneficial immunomodulation. (A) BIAcore SPR analysis of S. mutans adherence to immobilized salivary agglutinin. Sensograms show the binding of S. mutans to immobilized SAG in the presence of serum from BALB/c or C57/BL6 mice immunized with S. mutans alone or with IC containing MAbs 6-11A or 5-5D at high (saturating) or low (0.1× sub-saturating) concentrations. (B) Evaluation of IgG subclasses reactive with anti-S. mutans whole cells. Sera collected 7 days after primary immunization from BALB/c or C57/BL6 mice immunized with S. mutans alone (diamonds), IC containing MAb 6-11A at the higher concentration (squares), or lower concentration (triangles) were evaluated by quantitative ELISA. (C) Same as panel B except that MAb 5-5D was used in the experiment. (D) A competition ELISA was used to determine the level of MAb 1-6F-like Abs in the serum of BALB/c and C57/BL6 mice immunized with ICs of MAb 6-11A and 5-5D compared to S. mutans alone. The percent inhibition of binding of biotin-labeled 1-6F to S. mutans by sera from mice immunized with bacteria alone (white bars), high MAb concentration IC (black bars), and low MAb concentration IC (grey bars) is indicated. Results are expressed as mean ± SEM and statistical significance compared to serum from S. mutans only immunized mice is indicated by P values. Data are representative of at least three independent experiments. Statistical analysis was performed using graph pad prism 4.0 and analysis included one-way anova.

Article Snippet: Hence, optimal immunogenicity appears to represent a balance between preservation of sufficient conformational complexity to reconstitute relevant target epitopes, while disrupting the native structure such that immunodominance is shifted toward a more effective response. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 7 caption a7 Evaluation of immunogenic properties of recombinant P1 polypeptides. (A) BIAcore SPR analysis of S. mutans adherence to salivary agglutinin.

Techniques: Binding Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Labeling, Bacteria

Journal:

Article Title: Beneficial Immunomodulation by Streptococcus mutans anti-P1 Monoclonal Antibodies is Fc-independent and Correlates with Increased Exposure of a Relevant Target Epitope

doi: 10.4049/jimmunol.0803300

Figure Lengend Snippet: Evaluation of the role of activating Fc receptors in beneficial immunomodulation by anti-P1 MAbs. (A) BIAcore SPR analysis of S. mutans adherence to immobilized salivary agglutinin. Sensograms show the binding of S. mutans to immobilized SAG in the presence of serum from Fcer1g transgenic mice immunized with S. mutans alone or with IC containing high (saturating) or low (0.1× sub-saturating) concentrations of MAb 5-5D. (B) MAb 1-6F competition ELISA. The percent competition by sera from mice immunized with S. mutans alone (white bars), IC containing the high (saturating) MAb concentration (black bars), or IC containing the low (0.1× sub-saturating) MAb concentration (grey bars) is indicated. Results are expressed as mean ± SEM and statistical significance compared to serum from S. mutans only immunized mice is indicated by P values. (C) Quantitative anti-NR21 IgG subclass ELISA. Levels of anti-NR21 specific IgG1, IgG2a, and IgG2b subclass antibodies were measured in sera collected from Fcer1g transgenic mice immunized with S. mutans alone (diamonds), or with IC containing MAb 5-5D at the high saturating (squares) or low 0.1× sub-saturating concentration (triangles). Results are expressed as mean ± SEM and statistical significance compared to serum from S. mutans only immunized mice is indicated by P values. Data are representative of at least three independent experiments. Statistical analysis was performed using graph pad prism 4.0 and analysis included one-way anova.

Article Snippet: Hence, optimal immunogenicity appears to represent a balance between preservation of sufficient conformational complexity to reconstitute relevant target epitopes, while disrupting the native structure such that immunodominance is shifted toward a more effective response. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 7 caption a7 Evaluation of immunogenic properties of recombinant P1 polypeptides. (A) BIAcore SPR analysis of S. mutans adherence to salivary agglutinin.

Techniques: Binding Assay, Transgenic Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal:

Article Title: Beneficial Immunomodulation by Streptococcus mutans anti-P1 Monoclonal Antibodies is Fc-independent and Correlates with Increased Exposure of a Relevant Target Epitope

doi: 10.4049/jimmunol.0803300

Figure Lengend Snippet: Re-evaluation of the immunomodulatory properties of MAb 4-10A. (A) BIAcore SPR analysis of S. mutans adherence to salivary agglutinin in the presence of sera collected from BALB/c mice immunized with S. mutans alone or with ICs containing MAb 4-10A at the indicated dilutions. For the sake of clarity the change in resonance units detected over the 60 second injection cycle rather than the complete sensograms are shown. (B) MAb 1-6F competition ELISA. Sera collected from mice 7 days after primary immunization with S. mutans alone or with ICs containing MAb 4-10A at the indicated dilutions were tested for their ability to inhibit binding of biotin-labeled 1-6F to S. mutans whole cells. Results are expressed as the mean percent inhibition ± SEM and statistical significance compared to serum from S. mutans only immunized mice is indicated by P values. (C) Quantitative anti-NR21 IgG subclass ELISA. The levels of NR21-specific IgG1, IgG2a, and IgG2b subclass antibodies were measured in the same sera described in panel B. Results are expressed as mean ± SEM and statistical significance compared to serum from S. mutans only immunized mice is indicated by P values. (D) MAb 4-10A enhancement of MAb 1-6F binding to S. mutans whole cells by ELISA. The percent increase or decrease in binding of biotin-labeled MAb 1-6F to S. mutans in the presence of MAb 4-10A added at the indicated dilution was calculated. Data are representative of three independent experiments. Statistical analysis was performed using graph pad prism 4.0 and analysis included one-way anova.

Article Snippet: Hence, optimal immunogenicity appears to represent a balance between preservation of sufficient conformational complexity to reconstitute relevant target epitopes, while disrupting the native structure such that immunodominance is shifted toward a more effective response. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 7 caption a7 Evaluation of immunogenic properties of recombinant P1 polypeptides. (A) BIAcore SPR analysis of S. mutans adherence to salivary agglutinin.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Binding Assay, Labeling, Inhibition

Journal:

Article Title: Beneficial Immunomodulation by Streptococcus mutans anti-P1 Monoclonal Antibodies is Fc-independent and Correlates with Increased Exposure of a Relevant Target Epitope

doi: 10.4049/jimmunol.0803300

Figure Lengend Snippet: Evaluation of immunogenic properties of recombinant P1 polypeptides. (A) BIAcore SPR analysis of S. mutans adherence to salivary agglutinin. Sensograms show the binding of S. mutans to immobilized SAG in the presence of sera collected from sham-immunized (PBS) BALB/c mice compared to those immunized with GC14 (full-length recombinant P1) or with truncated polypeptides NR21, CK1 and CK2. (B) Sera collected from mice immunized with CG14 (filled diamonds), NR21 (open squares), CK1 (filled triangles), CK2 (filled squares), MBP alone (open diamonds), or acrylamide alone (open triangles) were tested for their total IgG reactivity against S. mutans whole cells by ELISA. (C) MAb 1-6F competition ELISA. The percent inhibition of biotin-labeled 1-6F to S. mutans whole cells by sera from mice immunized with CG14 (filled diamonds), NR21 (open squares), CK1 (filled triangles), CK2 (filled squares), MBP alone (open diamonds), or acrylamide alone (open triangles) are shown by line graph. The standard deviation between replicate samples was less than 5 percent in all cases.

Article Snippet: Hence, optimal immunogenicity appears to represent a balance between preservation of sufficient conformational complexity to reconstitute relevant target epitopes, while disrupting the native structure such that immunodominance is shifted toward a more effective response. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 7 caption a7 Evaluation of immunogenic properties of recombinant P1 polypeptides. (A) BIAcore SPR analysis of S. mutans adherence to salivary agglutinin.

Techniques: Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Labeling, Standard Deviation