Finegan et al master matrix 62 plots

see readme file

Numbers of Wa HRV-specific ASC per 5×10⁵ MNC obtained from gut-associated lymphoid tissues (Duodenum, Jejunum, Ileum) 21 PID.

<p>When comparing ASC numbers of the same isotype among treatment groups, different letters indicate a significant difference (Kruskal-Wallis rank sum test, p<0.05). n = number of piglets in each group. The IgM ASC response was not included as no IgM ASC were detected in most of the groups of pigs.</p

Geometric mean isotype-specific Ab titers (GMT) to Wa HRV per group per day (from ELISA assay) detected in rectal swab fluids of the experimental groups of piglets.

<p>All animals were orally inoculated at 72 h of age [0 post inoculation day (0 PID)], and euthanized at 21±3 PID. SIC: small intestinal contents; and LIC: large intestinal contents, were collected after euthanasia. The arrow at 0 PID indicates the inoculation day.</p

Immune responses to the passive treatments were determined in sera and intestinal contents of treated piglets.

<p>Continuous lines represent geometric mean of porcine IgG Ab titers (GMT) to the corresponding treatment administered (VHH, IgY or IgG Abs) in serum samples per group every 7 days. Dashed lines represent ELISA Ab titer of passive Abs to Wa HRV (VHH, IgY or IgG Abs) in rectal swabs every 7 days. The bars at PID 21 represent porcine IgG and IgA Abs to passive treatment in SIC: small intestinal contents and LIC: large intestinal contents collected after euthanasia. All animals were orally inoculated at 72 h of age [0 post inoculation day (0 PID)], and euthanized at PID 21±3. The arrow at PID 0 indicates the inoculation day. Different letters indicate significant differences between groups (p<0.05). In the case of groups CL, IgG and NT the results represent porcine IgG Ab titers to chicken IgY in serum but a similar response was observed against VHH nanoAbs (data not shown).</p

Diarrhea and virus shedding in Gn piglets after oral inoculation with VirHRV Wa (P[8]G1).

<p>Treatment Group abbreviations as well as passive treatments administered to each group of pigs are clarified in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003334#ppat-1003334-t001" target="_blank">Table 1</a>. VHH = piglets treated with VHH nanoAbs to VP6 at a final Wa HRV ELISA Ab titer of 4096; IgY = piglets treated with IgY Abs to VP6 at a final Wa HRV ELISA Ab titer of 4096; CL = piglets treated with control larvae; IgG = piglets treated with porcine IgG Abs to Wa HRV at a final Wa HRV ELISA Ab titer of 4096; NT = not treated piglets; n = number of animals per group; AUC = area under the curve. ND = not detected. PID = post inoculation day.</p>a<p>Determined by ELISA and CCIF.</p>b<p>Diarrhea duration was defined as the number of days with fecal score ≥2. Stool consistency was scored daily (0 = normal; 1 = pasty; 2 = semi-liquid; 3 = liquid).</p>c<p>Mean cumulative score was Σ(faecal scores ≥2)/number of animals. For statistical analysis, ND was considered as zero.</p>d<p>Proportions in the same column with different superscript upper case letters (A, B, C) differs significantly (Fisher's exact test, p<0.05).</p>e<p>Means in the same column with different superscript upper case letters differ significantly (Kruskall-Wallis rank sum test; p<0.05). Piglets were fed with 210 ml of sterile milk, twice a day, for 9 days with or without the addition of the corresponding volume of VP6 VHH pool, VP6 IgY pool, control larvae pool or Wa HRV porcine IgG pool, according to the experimental group.</p

Mean titer of virus shed per day per pig (from CCIF assay) detected daily in rectal swab fluids of the experimental groups of piglets.

<p>All animals were orally inoculated at 72 h of age [0 post inoculation day (0 PID)], and euthanized at 21±3 PID. SIC: small intestinal contents; and LIC: large intestinal contents, were collected after euthanasia. Horizontal bars represent the mean days of diarrhea duration for each group. The arrow at 0 PID indicates the inoculation day. The thin line indicates the duration of the passive treatments.</p

Numbers of Wa HRV-specific ASC per 5×10⁵ MNC obtained from systemic lymphoid tissues (blood and spleen) and MLN draining the small and large intestine at 21 PID.

<p>When comparing ASC numbers of the same isotype among treatment groups, different letters indicate a significant difference (Kruskal-Wallis rank sum test, p<0.05). n = number of piglets in each group. The IgM ASC response was not included as no IgM ASC were detected in most of the groups of pigs.</p

<i>In vitro</i> RVA fluorescent focus reduction assay using VHH antibodies.

<p>A fourfold dilution of each VHH clone (3B2 VHH, derived from llamas vaccinated with VP6 from RVA strain C486 -G6P[1]I?-,or non-related VHH nanoAb) was mixed with an equal volume of RVA containing 100 focus forming units (FFU) of each strain. The numbers represent the minimum VHH concentration that reduced >80% of the number of fluorescent focus forming units (FFU) of each RVA strain. Neg = no neutralizing activity was observed at the highest concentration tested (62.50 µg VHH/ml). The G-type, P-type and I-type of RVA strains is clarified in brackets next to each strain name (according to <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003334#ppat.1003334-Matthijnssens4" target="_blank">[91]</a>).</p