Journal: bioRxiv

Article Title: Antibody-based vaccine for tuberculosis: validation in horse foals challenged with the TB-related pathogen Rhodococcus equi

doi: 10.1101/292946

Figure Lengend Snippet: Serum end-point titers of IgG or IgG subisotypes are plotted by vaccine group as a function of age in days. ( A ) Total IgG antibody end-point titers to PNAG were significantly higher in sera of immunized mares at D21 and D0 PF compared with titers in sera of control mares at D0 PF. There was no significant (NS) difference in the IgG titers of the vaccinated mares at pre-immunization and controls at D0 PF. ( B-D ) Concentrations of IgG 1 , IgG 4/7 , and IgG 3/5 were significantly higher in mares in the vaccinated group at D21 and D0 PF as indicated on the figure. Statistical comparisons were made by linear mixed-effects regression with exchangeable correlation structure, using the mare as random effect (to account for repeated measures) and multiple comparisons determined by the method of Sidak.

Article Snippet: For IgG subisotype detection, 100 µl of goat-anti-horse IgG 4/7 (Lifespan Biosciences, Seattle, WA, diluted at 1:90,000), or goat anti-horse IgG 3/5 (Bethyl Laboratories, Montgomery, TX, USA, diluted at 1:30,000) conjugated to horseradish peroxidase, or mouse anti-horse IgG 1 (AbD Serotec, Raleigh, NC, USA), diluted at 1:25,000) were added to the wells and incubated for 1 hour at room temperature.

Techniques:

Journal: bioRxiv

Article Title: Antibody-based vaccine for tuberculosis: validation in horse foals challenged with the TB-related pathogen Rhodococcus equi

doi: 10.1101/292946

Figure Lengend Snippet: End-point colostral titers of IgG or IgG subisotype. End-point values are plotted by vaccine group. ( A ) Total IgG antibody end-point titers to PNAG were significantly higher in colostra of vaccinated mares (N=10, 2 samples not tested due to limited quantities) compared with colostra of control mares N=7). ( B ) Concentrations of IgG 1 , IgG 4/7 , and IgG 3/5 were significantly higher in colostra of mares in the vaccinated group (N=12). Statistical comparisons were made by the Wilcoxon rank-sum test.

Article Snippet: For IgG subisotype detection, 100 µl of goat-anti-horse IgG 4/7 (Lifespan Biosciences, Seattle, WA, diluted at 1:90,000), or goat anti-horse IgG 3/5 (Bethyl Laboratories, Montgomery, TX, USA, diluted at 1:30,000) conjugated to horseradish peroxidase, or mouse anti-horse IgG 1 (AbD Serotec, Raleigh, NC, USA), diluted at 1:25,000) were added to the wells and incubated for 1 hour at room temperature.

Techniques:

Journal: bioRxiv

Article Title: Antibody-based vaccine for tuberculosis: validation in horse foals challenged with the TB-related pathogen Rhodococcus equi

doi: 10.1101/292946

Figure Lengend Snippet: Endpoint serum titers (N=7 Non-vaccinated, 12 vaccinated) of IgG or IgG subisotypes are plotted by vaccine group as a function of age in days. ( A ) Total IgG antibody end-point titers to PNAG were significantly higher in an age-dependent matter between foals from mares that were vaccinated (filled symbols n=12) compared with titers in sera of foals from unvaccinated, control mares (open symbols n=7) through D39-42 of life. ( B-D ) Concentrations of IgG 1, IgG 4/7 , and IgG 3/5 were significantly higher in foals in the vaccinated group than the unvaccinated, control group through the day indicated on the figure. Statistical comparisons made using linear mixed-effects modeling with individual foal as a random effect; NS=not significant.

Article Snippet: For IgG subisotype detection, 100 µl of goat-anti-horse IgG 4/7 (Lifespan Biosciences, Seattle, WA, diluted at 1:90,000), or goat anti-horse IgG 3/5 (Bethyl Laboratories, Montgomery, TX, USA, diluted at 1:30,000) conjugated to horseradish peroxidase, or mouse anti-horse IgG 1 (AbD Serotec, Raleigh, NC, USA), diluted at 1:25,000) were added to the wells and incubated for 1 hour at room temperature.

Techniques:

Journal: bioRxiv

Article Title: Antibody-based vaccine for tuberculosis: validation in horse foals challenged with the TB-related pathogen Rhodococcus equi

doi: 10.1101/292946

Figure Lengend Snippet: ( A ) Titers of IgG subisotypes and IgA in control (open bars) and PNAG-immune (hatched bars) plasmas. ( B ) horse IgG 1 , ( C ) horse, IgG 3/5 , ( D ) horse IgG 4/7 , or ( E ) horse IgA in sera at day indicated on X-axis. Bars represent medians, error bars the interquartile ranges. OD ratios of IgG 1 , IgG 4/7 , and IgG 3/5 did not differ significantly over time in controls but were significantly (P < 0.05) greater than age 2 days in foals transfused with anti-PNAG plasma at ages 28, 42, and 56 days (IgG 1 ), or ages 28, 42, 56, and 84 for IgG 4/7 and IgG 3/5 . OD ratio of IgA did not differ significantly (P > 0.05) among the different days for anti-PNAG-transfused foals, but controls had significantly (P < 0.05) higher IgA titers at age 2 days compared to control titers on days 28, 42, and 56. The OD ratio values for control foals’ IgA on day 84 was significantly (P < 0.05) greater than those of control foals on days 28, 42, and 56. IgA titers between controls and anti-PNAG-transfused foals differed significantly (P < 0.05) at day 84 only. All P values were determined by mixed-effects linear regression. ( F ) Opsonic killing of R. equi EIDL 990 by antibody in control or immune plasma. Monoclonal antibodies (MAb) were used as controls, as were tubes lacking PMN or complement (C’) as indicated. Bars represent means of technical replicates.

Article Snippet: For IgG subisotype detection, 100 µl of goat-anti-horse IgG 4/7 (Lifespan Biosciences, Seattle, WA, diluted at 1:90,000), or goat anti-horse IgG 3/5 (Bethyl Laboratories, Montgomery, TX, USA, diluted at 1:30,000) conjugated to horseradish peroxidase, or mouse anti-horse IgG 1 (AbD Serotec, Raleigh, NC, USA), diluted at 1:25,000) were added to the wells and incubated for 1 hour at room temperature.

Techniques: