Journal: Microorganisms

Article Title: Virulence of Francisella tularensis Subspecies holarctica Biovar japonica and Phenotypic Change during Serial Passages on Artificial Media

doi: 10.3390/microorganisms8121881

Figure Lengend Snippet: Antigenic change in lipopolysaccharide (LPS) of KU-1 variants passaged on Eugon chocolate agar in Western blots. Purified LPSs of KU-1 original and the variants passaged on Eugon chocolate agar 10, 20, and 80 times (EP10, EP20, and EP80, respectively) and LVS (lanes 1, 2, 3, 4, and 5, respectively) were detected using MAbs M15C6, M13A13, and FB11 [sheets ( A ), ( B ), and ( C ), respectively]. M: molecular weight markers (Bio-Rad) indicate 75, 50, 37, 25, 20, 15, and 10 (arrow) kDa from the top.

Article Snippet: FB11 was purchased from Biodesign International (Saco, ME, USA).

Techniques: Chocolate, Western Blot, Purification, Molecular Weight

Journal: Immunology

Article Title: Protective B-cell epitopes of Francisella tularensis O -polysaccharide in a mouse model of respiratory tularaemia

doi: 10.1111/j.1365-2567.2012.03589.x

Figure Lengend Snippet: The epitope targeted by FB11 is contained within one OAg tetrasaccharide repeat whereas the epitope targeted by Ab52 spans two OAg tetrasaccharide repeats. Purified lipopolysaccharide (LPS) oligosaccharides were used to compete the binding of FB11 or Ab52 to Francisella tularensis OAgC coated on ELISA plates. Bars denote standard deviation of the mean from two or three experiments. –, 50% inhibition was not reached.

Article Snippet: Serially diluted purified oligosaccharides were pre-mixed with a fixed final concentration of 0·05 μg/ml FB11 or 3·125 μg/ml Ab52 in 2% MPBST in a Fisherbrand® 96-well round-bottom plate (Fisher Scientific, Pittsburgh, PA) at 1000 rpm for 1 hr at room temperature in an Eppendorf MixMate (Eppendorf AG, Hamburg, Germany).

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

Journal: Immunology

Article Title: Protective B-cell epitopes of Francisella tularensis O -polysaccharide in a mouse model of respiratory tularaemia

doi: 10.1111/j.1365-2567.2012.03589.x

Figure Lengend Snippet: FB11 and Ab52 confer survival to Francisella tularensis LVS-infected BALB/c mice in a dose-dependent manner. Five- to six-week old BALB/cJ female mice (n = 4–6) were inoculated intranasally with 5 × 104 to 7 × 104 colony-forming units (CFU) of LVS and injected intraperitoneally 1 hr later with graded doses of FB11 or Ab52. Survival was monitored for 28 days, and data from three experiments were combined. The log rank test was used to compare monoclonal antibody (mAb) -treated groups with vehicle groups. **P < 0·001; ***P < 0·0001. No difference in survival was observed between the groups treated with the 6 μg mAb dose or vehicle control.

Article Snippet: Serially diluted purified oligosaccharides were pre-mixed with a fixed final concentration of 0·05 μg/ml FB11 or 3·125 μg/ml Ab52 in 2% MPBST in a Fisherbrand® 96-well round-bottom plate (Fisher Scientific, Pittsburgh, PA) at 1000 rpm for 1 hr at room temperature in an Eppendorf MixMate (Eppendorf AG, Hamburg, Germany).

Techniques: Infection, Injection

Journal: Immunology

Article Title: Protective B-cell epitopes of Francisella tularensis O -polysaccharide in a mouse model of respiratory tularaemia

doi: 10.1111/j.1365-2567.2012.03589.x

Figure Lengend Snippet: FB11 and Ab52 reduce bacterial burden in BALB/c mice infected intranasally with Francisella tularensis LVS. Three groups of 19 BALB/cJ female mice, 5–6 weeks of age, were used in this experiment; seven mice were pre-assigned for survival (a) and the remaining mice were pre-assigned for determination of bacterial burden (b). Mice were inoculated intranasally with 7 × 104 colony-forming units (CFU) of LVS and 1 hr later, injected intraperitoneally with 50 μg FB11 or Ab52, or with PBS only. Survival was monitored for 28 days (a). For determination of bacterial burden, mice were euthanised on days 0 (1 hr after bacterial inoculation), 1, 3, 6, 14 and 28. For each time-point in B, three mice per group (except for the Ab52 group on day 14, when n = 2, and the PBS groups on days 14 and 28, when there were no surviving mice) were used for CFU determination, and means and standard deviations were calculated. Student’s t-test statistical analysis was performed and significant differences from the PBS control are indicated for each monoclonal antibody by color code: FB11, black, top; and Ab52, grey, bottom. *P < 0·05; **P < 0·005; ***P < 0·0005.

Article Snippet: Serially diluted purified oligosaccharides were pre-mixed with a fixed final concentration of 0·05 μg/ml FB11 or 3·125 μg/ml Ab52 in 2% MPBST in a Fisherbrand® 96-well round-bottom plate (Fisher Scientific, Pittsburgh, PA) at 1000 rpm for 1 hr at room temperature in an Eppendorf MixMate (Eppendorf AG, Hamburg, Germany).

Techniques: Infection, Injection

Journal: Immunology

Article Title: Protective B-cell epitopes of Francisella tularensis O -polysaccharide in a mouse model of respiratory tularaemia

doi: 10.1111/j.1365-2567.2012.03589.x

Figure Lengend Snippet: FB11 and Ab52, administered 2 hr or 19 hr after intranasal infection of BALB/c mice with Francisella tularensis SchuS4, prolong survival. BALB/cJ female mice, 7–8 weeks of age, were inoculated intranasally with SchuS4 and injected intraperitoneally with the indicated monoclonal antibody (mAb) doses or with Vehicle at the indicated times after infection. The log-rank test was used to compare mAb-treated groups with Vehicle-treated groups. P-values are indicated by colour code: FB11, black; Ab52, grey. (a) Combined results from four experiments for Ab52 [n = 22 (7 + 5+5 + 5)], only the first two of which included FB11 [n = 12 (7 + 5)]; the SchuS4 inocula for the four experiments were 20 colony-forming units (CFU), 25 CFU, 52 CFU and 57 CFU. (b) Ab52 at 30 or 100 μg (n = 5) tested in the same (57 CFU) experiment shown in (a). (c,d) FB11 and Ab52 at 10 μg (n = 5) or 50 μg (n = 5 for FB11, n = 10 for Ab52) tested 19 hr after infection with 35 CFU SchuS4.

Article Snippet: Serially diluted purified oligosaccharides were pre-mixed with a fixed final concentration of 0·05 μg/ml FB11 or 3·125 μg/ml Ab52 in 2% MPBST in a Fisherbrand® 96-well round-bottom plate (Fisher Scientific, Pittsburgh, PA) at 1000 rpm for 1 hr at room temperature in an Eppendorf MixMate (Eppendorf AG, Hamburg, Germany).

Techniques: Infection, Injection

Journal: Immunology

Article Title: Protective B-cell epitopes of Francisella tularensis O -polysaccharide in a mouse model of respiratory tularaemia

doi: 10.1111/j.1365-2567.2012.03589.x

Figure Lengend Snippet: FB11 and Ab52 reduce bacterial burden in BALB/c mice infected intranasally with Francisella tularensis SchuS4. Groups of BALB/c mice (n = 2 in a and 5 in b), 7–8 weeks of age, were infected intranasally with 116 colony-forming units (CFU) (a) or 164 CFU (b) of SchuS4 and injected intraperitoneally with 50 μg Ab52, FB11 or CO17-1A (IgG2a isotype control), 2 hr later. Mice were euthanised on day 3 after bacterial infection. Bacterial burdens in spleen and blood were determined by plating teased spleen suspension or heparinized blood on chocolate agar plates for bacterial enumeration (CFU per spleen or per ml blood). The difference between the Ab52 or FB11 and control (CO17-1A) groups was analysed by Student’s t-test. P < 0·05 is indicated by asterisk. Bars denote standard deviation from the means.

Article Snippet: Serially diluted purified oligosaccharides were pre-mixed with a fixed final concentration of 0·05 μg/ml FB11 or 3·125 μg/ml Ab52 in 2% MPBST in a Fisherbrand® 96-well round-bottom plate (Fisher Scientific, Pittsburgh, PA) at 1000 rpm for 1 hr at room temperature in an Eppendorf MixMate (Eppendorf AG, Hamburg, Germany).

Techniques: Infection, Injection, Standard Deviation