Journal: PLoS ONE

Article Title: Comparative Immunogenicity of 7 and 13-Valent Pneumococcal Conjugate Vaccines and the Development of Functional Antibodies to Cross-Reactive Serotypes

doi: 10.1371/journal.pone.0074906

Figure Lengend Snippet: Inhibition of serotype-specific 6A, 6B and 6C opsonophagocytic activity (GMTs) by purified polysaccharide following immunization with PCV7 or PCV13 vaccine.

Article Snippet: To assess the contribution of type specific antibody to killing of cross-reactive antigens, sera were retested by OPA after adsorbing sera with purified 19F, 19A, 6A, 6B or 6C capsular polysaccharide (ATCC for 19F and 19A and Statens Serum Institute for 6A, 6B, and 6C) one at a time and reported as the titer where 50% of bacteria were killed.

Techniques: Inhibition, Activity Assay, Purification

Journal: Nature Communications

Article Title: Dynamic capsule restructuring by the main pneumococcal autolysin LytA in response to the epithelium

doi: 10.1038/ncomms10859

Figure Lengend Snippet: ( a ) Schematic representation of localization of cell wall synthesis and autolysin (red=capsule, green=old cell wall, black=new cell wall containing choline, white=new cell wall containing ethanolamine, LytA=black diamonds). ( b ) TIGR4 was grown as described for ethanolamine pulse experiments (Methods). Penicillin was added as indicated at 40 ng ml −1 . For an autolytic negative reference the mutant in lytA (LytA − ) grown in choline was included. ( c ) Cultures from b after ethanolamine or choline pulse were harvested and used in a standard capsule-shedding assay and the supernatant fractions were analysed by capsule blot. Std: purified capsule polysaccharides. ( d ) TIGR4 was subjected to a capsule-shedding assay with either LL-37 at 4 μg ml −1 , pencillin (10–80 ng ml −1 ) or vancomycin (100–800 ng ml −1 ). Supernatants from these assays were then analysed by capsule blot. Std: purified capsule polysaccharides. ( e ) At the arrows, cultures of TIGR4 were exposed to either penicillin (Pen) or vancomycin (Vanc) at the indicated concentrations and growth was monitored by optical density.

Article Snippet: Past methods of polyacrylamide gel electrophoresis (PAGE) resulted in <5% of capsule entering the resolving portion of the gel, and the low molecular weight species from bacterial samples that entered PAGE gels were not apparent in commercially available purified capsular polysaccharides (ATCC, serotype 4 cat 173-X, serotype 3 cat 169-X, serotype 2 cat 165-X, serotype 6A cat 271-X and serotype 19F cat 99-X).

Techniques: Mutagenesis, Purification

Journal: Frontiers in Immunology

Article Title: Pneumococcal Polysaccharide Vaccination Elicits IgG Anti-A/B Blood Group Antibodies in Healthy Individuals and Patients with Type I Diabetes Mellitus

doi: 10.3389/fimmu.2016.00493

Figure Lengend Snippet: IgM/IgG antibody response to pneumococcal polysaccharides after immunization with Pn23 vaccine . Healthy individuals ( n = 28) and patients with type I DM ( n = 16) were immunized with the PnPs vaccine Pneumo 23 Vaccine “Pasteur Merieux” (Pn23), and blood samples were drawn 4–6 weeks after vaccination. Antibodies against pneumococcal polysaccharides (anti-PnPs) were determined by ELISA as described in Section “ ” and presented as IgM–Pn23-antibody response (A) or IgG–Pn23-antibody response (B) . Results are depicted using box plot diagrams, with the median represented by a cross, the interquartile range (IQR) represented by the box, and 5- and 95-percentile values represented by the whiskers; statistically significant differences between study groups were calculated using the non-parametric two-tailed Mann–Whitney U -test.

Article Snippet: During a clinical study published previously , healthy individuals ( n = 39) and patients with type I DM ( n = 20) were immunized with the PnPs vaccine Pn23 (a 23-valent vaccine containing pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6B, 7, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F, Pasteur Merieux Connaught, Lyon, France) after informed consent was obtained ( , ).

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Pneumococcal Polysaccharide Vaccination Elicits IgG Anti-A/B Blood Group Antibodies in Healthy Individuals and Patients with Type I Diabetes Mellitus

doi: 10.3389/fimmu.2016.00493

Figure Lengend Snippet: Increase in blood group IgG anti-A/B antibodies in healthy individuals and in patients with type I DM after immunization with Pn23 vaccine as determined with the Biacore ® device . To determine IgM (A) and IgG (B) anti-A/B antibodies in healthy individuals and patients with type I DM prior and post immunization with Pneumo 23 Vaccine “Pasteur Merieux” (Pn23), an anti-human IgM mAb (A) or anti-human IgG Abs (B) were applied as the second step reagents as described in Section “ .” Results are depicted using box plot diagrams, with the median represented by a cross, the interquartile range (IQR) represented by the box, and 5- and 95-percentile values represented by the whiskers; statistically significant differences between study groups were calculated using the non-parametric two-tailed Mann–Whitney U -test. For the IgM antibody response, no statistically significant differences were found between study groups.

Article Snippet: During a clinical study published previously , healthy individuals ( n = 39) and patients with type I DM ( n = 20) were immunized with the PnPs vaccine Pn23 (a 23-valent vaccine containing pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6B, 7, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F, Pasteur Merieux Connaught, Lyon, France) after informed consent was obtained ( , ).

Techniques: Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Pneumococcal Polysaccharide Vaccination Elicits IgG Anti-A/B Blood Group Antibodies in Healthy Individuals and Patients with Type I Diabetes Mellitus

doi: 10.3389/fimmu.2016.00493

Figure Lengend Snippet: Binding characteristics of anti-A/B antibodies after immunization with Pn23 vaccine . The percentage of dissociation (A) of blood group anti-A/B antibodies before and after immunization with Pneumo 23 Vaccine “Pasteur Merieux” (Pn23) in healthy individuals ( n = 28) and in patients with type I DM ( n = 16) was investigated using the Biacore device, the CM5 chip, and the synthetic blood group specific A/B trisaccharides as described in Section “ .” Furthermore, the estimated affinity constant K D (B) of blood group specific anti-A antibodies before and after pneumococcal vaccination were analyzed in serum samples of three healthy individuals and two patients with type I DM selected for their initially high anti-A antibody titer (>than 1:256 or >than 700 RU). Kinetic measurements of anti-A antibodies binding to surface bound trisaccharide before and after vaccination in two representative individuals (a healthy individual, Ind1, and a DM I patient, Ind2) who fulfilled these criteria are given; one healthy individual that was not included in the vaccination protocol but showed high titer for blood group anti-A antibodies was measured in parallel (Control). Green curves represent serum sample and red curves serum sample diluted by half; the black curves depict the calculated fit. n.d., not detectable.

Article Snippet: During a clinical study published previously , healthy individuals ( n = 39) and patients with type I DM ( n = 20) were immunized with the PnPs vaccine Pn23 (a 23-valent vaccine containing pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6B, 7, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F, Pasteur Merieux Connaught, Lyon, France) after informed consent was obtained ( , ).

Techniques: Binding Assay, Control