Journal:

Article Title: Invasion and Killing of Human Endothelial Cells by Viridans Group Streptococci

doi: 10.1128/IAI.71.5.2365-2372.2003

Figure Lengend Snippet: Bacterial species and strains used in this study and their invasion of HUVEC monolayers

Article Snippet: Streptococcus gordonii strain CH1 (Challis) was chosen as the paradigm for most of the experiments reported here because it is virulent in the rat model of infection ( 59 ), several adhesion-deficient mutants were available for comparison ( 14 , 39 , 40 ), and it produces large amounts of the alpha-hemolysin (hydrogen peroxide) ( 3 , 4 ). table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Bacterium Description Source or reference HUVEC invasion a S. gordonii DL1 (Challis) Wild-type reference strain 52 17 ± 5 EM230 DL1 hsa :: ermAM 52 4 ± 2 OB219 DL1 sspA ′ sspB ′:: ermAM 14 20 ± 4 OB220 DL1 sspA :: ermAM 14 19 ± 6 OB235 DL1 cshA3 :: ermAM 40 2 ± 0.4 OB271 DL1 cshB2 :: ermAM 40 2 ± 1 OB277 DL1 cshA31 :: cat cshB2 :: ermAM 40 2 ± 0.6 CH1 (Challis) Wild-type reference strain 59 82 ± 15 AMS12 CH1 with 1.7-kb Hin d fragment of gtfG replaced with lacZ /Erm r 59 14 ± 0.5 M5 Wild type from human dental plaque 28 5 ± 2.5 S. sanguis L22 Wild type from human dental plaque 28 76 ± 11 L52 Wild type from human dental plaque 28 9 ± 2 L74 Wild type from human dental plaque 28 52 ± 8 L79 Wild type from human dental plaque 28 14 ± 5 133-79 Wild type from human endocarditis 28 21 ± 5 2017-78 Wild type from human endocarditis 28 25 ± 3 10556 Wild type from human endocarditis ATCC 98 ± 18 S. oralis KS32AR Wild type from dental plaque 8 9 ± 8 16532AR Wild type from dental plaque 8 3 ± 2 S. mitis NCTC 10712 Wild type from dental plaque 8 23 ± 6 OP51 Wild type from dental plaque 8 35 ± 9 S. mutans UA159 Wild type from dental plaque 60 26 ± 7 S. salivarius 13419 Wild type from human saliva ATCC 2 ± 1 E. coli HB101 Hybrid of strains K-12 and B, transformable strain 7 <0.01 Open in a separate window a Average number of intracellular bacteria (× 10 4 ) per HUVEC monolayer ± standard deviation of the mean ( n = 6). b ATCC, American Type Culture Collection.

Techniques:

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Plasmid Preparation, Cloning

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: S. gordonii strains differ in adherence following neuraminidase pretreatment. The graph shows adherence of S. gordonii strains over 1 h to oral epithelial cell line TR146 pretreated with either purified neuraminidase (N) or medium alone for 30 min. Adherence is expressed as a percentage of the inoculum. Values are the means for at least three independent experiments, each performed in triplicate, ±SD. Asterisks indicate a significant difference in adherence of the S. gordonii strain to neuraminidase-treated cells compared to the tissue culture medium control. Statistical significance was tested by two-tailed Student's t test. *, P < 0.05.

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Purification, Control, Two Tailed Test

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: Serine-rich repeat proteins Hsa and GspB are required for binding to sialic acid but not to underlying carbohydrates. The graphs show adherence of S. gordonii strains DL1 (A), PS478 (B), and SK186 (C) and isogenic serine-rich repeat protein mutants to oral epithelial cell line TR146 pretreated with either neuraminidase (N) or medium alone for 30 min. Adherence after an hour of incubation was expressed as a percentage relative to the untreated parental strain. Values are the means for at least three independent experiments, each performed in triplicate, ±SD. Statistical significance was tested by ANOVA with an LSD post hoc test. *, P < 0.05. NS, not significant.

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Binding Assay, Incubation

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: Some S. gordonii strains bind β-1,4-linked galactose on oral epithelial cells. The graphs show adherence of S. gordonii DL1 (A), PS478 (B), and SK186 (C), in the presence of SpBgaA146-990 (B) or PBS control, to oral epithelial cell line TR146 pretreated with either neuraminidase (N) or medium alone. Adherence after an hour of incubation was expressed as a percentage relative to the untreated strain. Values are the means for at least three independent experiments, each performed in triplicate, ±SD. Statistical significance was tested by ANOVA with an LSD post hoc test. *, P < 0.05. NS, not significant.

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Control, Incubation

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: Some S. gordonii strains bind β-1,4-linked galactose on pharyngeal epithelial cells. The graphs show adherence of S. gordonii to D562 cells pretreated with neuraminidase in the presence or absence of SpBgaA146-990 (B). Adherence after an hour was expressed as a percentage relative to the untreated strain. Values are the means for at least three independent experiments, each performed in triplicate, ±SD. Statistical significance was tested by two-tailed Student's t test. *, P < 0.05. NS, not significant.

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Two Tailed Test

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: S. gordonii binding to β-1,4 galactose is not mediated by BgaA. The graphs show adherence of S. gordonii strains PS478 and SK186 and isogenic bgaA (A) and bgaA gspB (B) mutants to neuraminidase-treated (N) oral epithelial cell line TR146. Adherence after an hour of incubation was expressed as a percentage relative to the untreated parental strain. Values are the means for at least three independent experiments, each performed in triplicate, ±SD. Statistical significance was tested by ANOVA with an LSD post hoc test *, P < 0.05. NS, not significant.

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Binding Assay, Incubation

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: S. gordonii binding to β-1,4 galactose is primarily sortase A mediated. The graphs show adherence of S. gordonii strains PS478 (A) and SK186 (B) and isogenic serine-rich repeat protein mutants, in the presence of SpBgaA146-990 (B) or PBS control, to oral epithelial cell line TR146 pretreated with either neuraminidase (N) or medium alone. Adherence after an hour of incubation was expressed as a percentage relative to the untreated parental strain. Values are the means for at least three independent experiments, each performed in triplicate, ±SD. Statistical significance was tested by ANOVA with an LSD post hoc test. *, P < 0.05. NS, not significant.

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Binding Assay, Control, Incubation

Journal: Infection and Immunity

Article Title: Role of Neuraminidase-Producing Bacteria in Exposing Cryptic Carbohydrate Receptors for Streptococcus gordonii Adherence

doi: 10.1128/IAI.00068-18

Figure Lengend Snippet: S. oralis can increase adherence of S. gordonii SK186 in a neuraminidase-dependent manner. The graph shows adherence of the S. gordonii SK186 ΔbgaA strain to the oral epithelial cell line TR146 in the presence of neuraminidase (N), the S. oralis ATCC 10557 ΔbgaA (So ΔbgaA) or S. oralis ATCC 10557 ΔbgaA ΔnanA (So ΔbgaA ΔnanA) strain, or medium alone. Adherence after an hour of incubation was expressed as a percentage relative to the untreated parental strain. Values are the means for at least three independent experiments, each performed in triplicate, ±SD. Statistical significance was tested by ANOVA with an LSD post hoc test. *, P < 0.05. NS, not significant.

Article Snippet: This allows studies to focus on the differences due to mutations or treatments without the distraction of variability in baseline adherence between experiments; this strategy is often used when studying streptococcal adherence to epithelial cells ( 32 , – 35 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Characteristic(s)/genotype c Source or reference Strains Streptococcus gordonii DL1 S. gordonii Challis 66 b DL1 Δ hsa Δ hsa :: aad9 , Spc r This study SK12 S. gordonii oral isolate 67 b 38 Human isolate 68 b SK33 S. gordonii oral isolate 67 b PS478 S. gordonii endocarditis isolate 17 PS478 Δ gspB Δ gspB :: aad9 , Spc r This study PS478 Δ bgaA Δ bgaA :: aad9 , Spc r This study PS478 Δ srtA Δ srtA :: aad9 , Spc r This study PS478 Δ bgaA Δ gspB This study G9B S. gordonii oral isolate 69 b M99 S. gordonii endocarditis isolate 70 ATCC 10558 S. gordonii endocarditis isolate 66 b SK186 a S. gordonii oral isolate 71 b SK186 Δ gspB Δ gspB :: aad9 , Spc r This study SK186 Δ bgaA Δ bgaA :: aad9 , Spc r This study SK186 Δ srtA Δ srtA :: aad9 , Spc r This study SK186 Δ bgaA Δ gspB This study 72-40 72 b Streptococcus oralis ATCC 10557 S. oralis endocarditis isolate 73 b ATCC 10557 Δ bgaA Δ bgaA :: kan-rpsL + , Kan r This study ATCC 10557 Δ bgaA ΔnanA Δ bgaA :: kan-rpsL + Δ nanA :: erm , Erm r Kan r This study Escherichia coli strain Stellar Cloning host Clontech Plasmids pDrive Cloning vector, Amp r Kan r Qiagen pDriveΔgspB pDriveΔ gspB :: aad9 , Spc r Kan r This study pDriveΔ bgaA pDriveΔ bgaA :: aad9 , Spc r Amp r Kan r This study pDriveΔ srtA pDriveΔ srtA :: aad9 , Spc r Kan r This study pDriveΔ nanA pDriveΔ nanA :: erm , Erm r Amp r Kan r 25 pJET1.2/blunt Cloning vector, Amp r Thermo Fisher Scientific pJET SobgaA pJET1.2 containing a fragment of ATCC 10557 bgaA This study pJET So Δ bgaA pJET SobgaA with Janus cassette, Amp r Kan r This study pJET gspB pJET1.2 containing a fragment of 7240 gspB This study pJETΔ gspB pJET gspB with Janus cassette, Amp r Kan r This study Open in a separate window a Previously named LGR2. b These strains were initially identified as different streptococcal species but were reclassified into S. gordonii . c Kan r , kanamycin resistant; Amp r , ampicillin resistant; Spc r , spectinomycin resistant; Erm r , erythromycin resistant.

Techniques: Incubation

Journal:

Article Title: Invasion and Killing of Human Endothelial Cells by Viridans Group Streptococci

doi: 10.1128/IAI.71.5.2365-2372.2003

Figure Lengend Snippet: Bacterial species and strains used in this study and their invasion of HUVEC monolayers

Article Snippet: Streptococcus gordonii strain CH1 (Challis) was chosen as the paradigm for most of the experiments reported here because it is virulent in the rat model of infection ( 59 ), several adhesion-deficient mutants were available for comparison ( 14 , 39 , 40 ), and it produces large amounts of the alpha-hemolysin (hydrogen peroxide) ( 3 , 4 ). table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Bacterium Description Source or reference HUVEC invasion a S. gordonii DL1 (Challis) Wild-type reference strain 52 17 ± 5 EM230 DL1 hsa :: ermAM 52 4 ± 2 OB219 DL1 sspA ′ sspB ′:: ermAM 14 20 ± 4 OB220 DL1 sspA :: ermAM 14 19 ± 6 OB235 DL1 cshA3 :: ermAM 40 2 ± 0.4 OB271 DL1 cshB2 :: ermAM 40 2 ± 1 OB277 DL1 cshA31 :: cat cshB2 :: ermAM 40 2 ± 0.6 CH1 (Challis) Wild-type reference strain 59 82 ± 15 AMS12 CH1 with 1.7-kb Hin d fragment of gtfG replaced with lacZ /Erm r 59 14 ± 0.5 M5 Wild type from human dental plaque 28 5 ± 2.5 S. sanguis L22 Wild type from human dental plaque 28 76 ± 11 L52 Wild type from human dental plaque 28 9 ± 2 L74 Wild type from human dental plaque 28 52 ± 8 L79 Wild type from human dental plaque 28 14 ± 5 133-79 Wild type from human endocarditis 28 21 ± 5 2017-78 Wild type from human endocarditis 28 25 ± 3 10556 Wild type from human endocarditis ATCC 98 ± 18 S. oralis KS32AR Wild type from dental plaque 8 9 ± 8 16532AR Wild type from dental plaque 8 3 ± 2 S. mitis NCTC 10712 Wild type from dental plaque 8 23 ± 6 OP51 Wild type from dental plaque 8 35 ± 9 S. mutans UA159 Wild type from dental plaque 60 26 ± 7 S. salivarius 13419 Wild type from human saliva ATCC 2 ± 1 E. coli HB101 Hybrid of strains K-12 and B, transformable strain 7 <0.01 Open in a separate window a Average number of intracellular bacteria (× 10 4 ) per HUVEC monolayer ± standard deviation of the mean ( n = 6). b ATCC, American Type Culture Collection.

Techniques:

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Retooling a Blood-based Biomarker: Phase I Assessment of the High-Affinity CA19-9 Antibody HuMab-5B1 for Immuno-PET Imaging of Pancreatic Cancer

doi: 10.1158/1078-0432.CCR-18-3667

Figure Lengend Snippet: A, There is a reduction in liver uptake of radioactivity with increasing antibody mass (error bars show one standard deviation). B, Maximum-intensity projection image demonstrates the effect in a patient injected with 3 mg antibody (left) and a (different) patient injected with 47 mg HuMab 5B1 antibody 2 h before injection of 3 mg radiolabeled HuMab 5B (right). C, Fused PET/CT, PET, and contrast-enhanced CT of PDAC liver metastases in a patient who received 47 mg of HuMab 5B1 before injection of the radiolabeled antibody. Small liver metastases measuring less than 5 mm (red arrows) are visualized with high contrast. SUVs values were 101.4, 70.7, and 53.4 g/ml, respectively. Arrowhead indicates splenic infarct.

Article Snippet: The fully human HuMab-5B1 antibody was manufactured for MabVax Therapeutics Holdings, Inc. (San Diego, CA) by Patheon Biologics (Groningen, Netherlands) under current good manufacturing practice (cGMP) guidelines and supplied in sterile vials to MSK.

Techniques: Radioactivity, Standard Deviation, Injection, Positron Emission Tomography-Computed Tomography