Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Monoclonal antibodies isolated from mice immunized with CP5 or CP8 conjugate vaccines.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: Isolation

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Reactivity of five mAbs with native and O-deacetylated CP5 and CP8. (A) Structural similarities between CP5 and CP8 and their sites of O-acetylation (O-Ac). Both CP5-specific mAbs (4C2 and 5D1) reacted with (B) native CP5, whereas only mAb 4C2 reacted with (C) O-deacetylated CP5 (De-O-CP5). Three CP8-specific mAbs (5A6, 3B5, and 4G5) reacted with (D) native CP8, whereas only 5A6 and 3B5 reacted with (E) O-deacetylated (De-O-CP8) CP8. D-ManNAcA, N-acetyl-D-mannosaminuronic acid; L-FucNAc, N-acetyl-L-fucosamine; D-FucNAc, N-acetyl-D-fucosamine. Each sample was tested in duplicate, and the ELISAs were performed at least twice. The mean values of a representative experiment are shown.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques:

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Strains used in this study or tested in the colony immunoblot.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: Western Blot, Genetically Modified, Expressing, Mutagenesis

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: mAbs CP5–4C2 and CP8–5A6 were tested by colony immunoblot for reactivity with S. aureus type 5 and 8 reference strains or clinical isolates. The staphylococcal isolates are listed in Table 2. mAb 4C2 reacted with all 14 serotype 5 isolates, whereas mAb 5A6 reacted with 19 of 21 serotype 8 isolates. CoNS, coagulase-negative staphylococci; CP-, capsule negative. The immunoblot was performed three times, and a representative blot is shown.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: Western Blot

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Quantitation of cell-associated and soluble CP5 or CP8 produced by clinical isolates of S. aureus cultivated in RMPI + 1% casamino acids. CP levels were measured by an ELISA inhibition method, and CP5 and CP8 concentrations are expressed relative to those of Reynolds (CP5) and Reynolds (CP8), respectively, which were included as reference strains on every assay. CP levels were compared by one-way ANOVA, and multiple comparisons were made to the reference strains Reynolds (CP5) and Reynolds (CP8). ***, P < 0.001; **, P < 0.01; *, P < 0.05.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: Quantitation Assay, Produced, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Opsonic activity of CP5-specific mAbs (A) 4C2 and (B) 5D1 against wild type strain Reynolds (CP5) and its isogenic cap5H deletion mutant that produces wild-type levels of O-deacetylated CP5. The mAbs were serially diluted 3-fold and tested in the OPK assay. The results are expressed as percent changes in the number of CFU/ml after a 2-h incubation with mAb, HL60 cells, and a complement (C’) source. The samples labeled SA+ C’+HL60 contain no mAb. The titer is expressed as the lowest mAb dilution that killed 50% of the inoculum (dotted line). The experiment was performed twice, and a representative experiment is shown.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: Activity Assay, Mutagenesis, Incubation, Labeling

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Comparative opsonic activity of mAbs against serotype 5 and 8 S. aureus strains. (A) Backbone-specific CP5 mAb 4C2 showed somewhat better opsonic activity against Reynolds (CP5) than strain Newman. (B) Backbone-specific CP8 mAbs 5A6 showed comparable opsonic activity against Reynolds (CP8) and MRSA strain ST80–16. (C) O-acetyl-specific CP8 mAb 4G5 showed greater OPK activity against Reynolds (CP8) than strain ST80–16. The sample labeled 0 antibody contained HL60s, S. aureus, and the complement source. The data shown are means ( ± standard errors) of three to four experiments.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: Activity Assay, Labeling

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Passive immunization with CP5- or CP8-specific mAbs in the mouse bacteremia model. Mice were immunized IV with 100 µg of (A) CP5 mAb 4C2 or (B) CP8 mAb 5A6 24 h before challenge by the IP route with 107 CFU (A) S. aureus Reynolds (CP5) or (B) Reynolds (CP8). Control mice were immunized with irrelevant mAb 6C5 or polyclonal CP5 specific rabbit IgG (300 µg), and all mice were bled 2 h after bacterial challenge. The horizontal lines represent median CFU/ml blood for each group of mice. Bacteremia levels in mice administered mAbs 4C2 or 5A6 were compared by the Mann-Whitney U test to levels in control mice given mAb 6C5. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: MANN-WHITNEY

Journal: Virulence

Article Title: Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo

doi: 10.1080/21505594.2016.1270494

Figure Lengend Snippet: Plasma levels of CP5 or CP8 in mice infected IV with ∼2 × 108 CFU of S. aureus NRS382 (CP5+) or ST80–17 (CP8+), respectively. After 1 to 5 days, the animals were bled by cardiac stick, and the plasma extracts were prepared. CP levels were determined by an ELISA inhibition assay, and CP concentrations were compared by the Mann-Whitney U test. *, P < 0.05. The lower limit of CP detection is indicated by a dashed line.

Article Snippet: A3 S. epidermidis RP62A NA Biofilm producing, methicillin-resistant strain 65 A4 S. haemolyticus JCSC 1435 NA Sequenced strain from a Japanese patient in Tokyo 66 A5 S. saprophyticus 15305 NA ATCC; genome sequenced uropathogenic isolate 67 A6 S. lugdunensis 45–6447 NA Bacteremia isolate from 2014 BWH a Staphylococcus aureus B1 Newman 5 Human isolate from 1952 68 B2 Mu50 5 Vancomycin-intermediate MRSA isolate; Japan 69 B3 COL 5 MRSA strain with SCC mec II 65 B4 502A 5 Strain adept at colonization 70 B5 NRS 382 5 USA100 MRSA NARSA b B6 NRS 383 5 USA200 MRSA NARSA B7 NRS 386 5 USA700 MRSA NARSA C1 Lowenstein 5 Nabi serotype 5 reference strain 71 C2 Reynolds (CP5) 5 Prototype CP5+ strain 17 C3 Reynolds Δ cap5H 5 Produces WT levels of O-deacetylated CP5 51 C4 Crimson 5313 5 Bacteremia isolate from 2012 BWH C5 Crimson 18207 5 Bacteremia MRSA isolate from 2013 BWH C6 29213 5 Antibiotic susceptibility testing reference strain ATCC C7 ST22 #15 5 CA-MRSA c from 2010; Regensburg, Germany F. Hanses D1 MN8 8 Menstrual toxic shock isolate 72 D2 ST80–16 8 CA-MRSA strain from 2010; Regensburg, FRG 73 D3 Wright 8 Nabi serotype 8 reference strain 71 D4 PS80 8 ATCC 27700 58 D5 Becker 8 Prototype CP8+ strain 74 D6 V8 8 Source of staphylococcal V8 protease 75 D7 HS-522 8 ST239 MRSA 76 E1 NRS 387 8 USA800 MRSA NARSA E2 NRS 483 8 USA1000 MRSA NARSA E3 NRS 484 8 USA1100 MRSA NARSA E4 GFRY 8 Bacteremia isolate from 1997 BWH E5 VP 8 Bacteremia isolate from 1995 41 E6 JP122 8 Bacteremia isolate from 1990 BWH E7 JP136 8 Bacteremia isolate from 1990 BWH F1 25923 8 Seattle 1945; antibiotic reference strain ATCC F2 Reynolds (CP8) 8 Reynolds genetically modified to produce CP8 17 F3 UAMS-1 8 Genome sequenced musculoskeletal isolate 77 F4 2672 8 USA400; necrotizing pneumonia and sepsis isolate 32 F5 RF122 8 Genome sequenced bovine isolate 78 F6 Sanger 252 8 Epidemic MRSA clone 31 F7 MW2 8 Community acquired MRSA; USA400 79 G1 Sanger 476 CP- Community acquired MSSA; USA400 31 G2 8325–4 CP- NCTC; cured of three prophages 80 G3 LAC CP- Los Angeles clone; USA300 MRSA 81 G4 NRS 482 CP- USA300 MRSA NARSA G5 NRS 691 CP- USA500 MRSA NARSA G6 Cowan I CP- Isolate with high protein A expression 82 G7 Reynolds (CP-) CP- Reynolds acapsular mutant 17 Open in a separate window Note . a BWH, Brigham and Women's Hospital, Boston, MA. b NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus. c CA-MRSA, community-acquired methicillin resistant Staphylococcus aureus.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Inhibition, MANN-WHITNEY