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## Results: 7

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Intestinal numbers of 2/4/6/7VLP-GFP

^{+}plasmacytoid dendritic cells (pDCs), conventional dendritic cells (cDCs) and macrophages/monocytes after human rotavirus (HRV) infection. (a) HRV-inoculated and (b) medium-alone-inoculated representative dot plots of SWC3 versus CD4 at post-infection day (PID) 2, 4 and 7 are shown. Gates on SWC3^{low}CD4^{+}DCs (pDCs), SWC3^{high}CD4^{−}DCs (macrophages/monocytes) and SWC3^{low}CD4^{−}DCs (cDCs) and plots on each gate show 2/4/6/7VLP-GFP^{+}DCs. Numbers of 2/4/6/7VLP-GFP^{+}pDCs (c), 2/4/6/7VLP-GFP^{+}cDCs (d) and 2/4/6/7VLP-GFP^{+}macrophages/monocytes (e) in piglets that received HRV (○) or medium alone (▪) are summarized. For each experiment 100 000 events were acquired by the flow cytometer. The number of piglets studied at each PID is shown at the bottom of each graph. The error bars denote the standard error of the mean (SEM) and bars with an asterisk differ significantly from the controls at each PID (Kruskall–Wallis rank sum test*P*≤ 0.05).3.

Intestinal frequencies of interferon-α (IFN-α) positive plasmacytoid dendritic cells (pDCs), conventional dendritic cells (cDCs) and macrophages/monocytes after human rotavirus (HRV) infection. (a) HRV inoculated and (b) medium alone inoculated representative dot plots of SWC3 versus CD4 at post-infection days (PID) 2, 4 and 7 are shown. Gates on SWC3

^{low}CD4^{+}DCs (pDCs), SWC3^{high}CD4^{−}DCs (macrophage/monocytes) and SWC3^{low}CD4^{−}DCs (cDCs) and plots on each gate show IFN-α^{+}DCs. Mean frequencies of IFN-α^{+}pDCs (c), IFN-α^{+}cDCs (d) and IFN-α^{+}macrophages/monocytes (e) in piglets that received HRV (○) or medium alone (▪) are summarized. For each experiment 100 000 events were acquired by the flow cytometer. The number of piglets studied at each PID is shown at the bottom of each graph. The error bars denote the standard error of the mean (SEM) and bars with an asterisk differ significantly from the controls at each PID (Kruskall–Wallis rank sum test*P*≤ 0.05).4.

Total intestinal frequencies of plasmacytoid dendritic cells (pDCs), conventional dendritic cells (cDCs) and macrophages/monocytes after human rotavirus (HRV) infection. (a) HRV-inoculated piglets and (b) medium-inoculated piglets, representative dot plots of SWC3 versus CD4 at post-infection day (PID) 2, 4 and 7 and isotype control plots are shown. Gates and percentages of SWC3

^{low}CD4^{+}DCs (pDCs), SWC3^{high}CD4^{−}DCs (macrophages/monocytes) and SWC3^{low}CD4^{−}DCs (cDCs) are shown. Mean total frequencies of intestinal pDCs (c), macrophages/monocytes (d) and cDCs (e) in piglets that received HRV (○) or media alone (▪) are summarized. For each experiment 100 000 events were acquired by the flow cytometer. The number of piglets studied at each PID is shown at the bottom of each graph. The error bars denote the standard error of the mean (SEM) and time-points with an asterisk differ significantly from the controls at each PID (Kruskall–Wallis rank sum test*P*≤ 0.05).5.

Serum and small intestinal content (SIC) cytokines after human rotavirus (HRV) infection. Serum and SIC cytokines were measured by enzyme-linked immunosorbent assay. Graphs to the left represent serum cytokines and graphs to the right represent SIC cytokines at post-infection days (PID) 2, 4 and 7 from top to bottom: interferon-α (IFN-α), interleukin-12 (IL-12), tumour necrosis factor-α (TNF-α), IL-6 and IL-10. Open circles represent the cytokine amount in serum or SIC in HRV-infected piglets and black squares represent cytokine levels in serum or SIC in mock-inoculated piglets and at day 0. The number of piglets used per PID is shown at the bottom of each graph. The error bars denote the standard error of the mean (SEM) and time-points with a bar and an asterisk differ significantly from the controls at each PID noted (Kruskall–Wallis rank sum test

*P*≤ 0.05).6.

Diarrhoea scores, percentage of piglets with jejunal pathology, serum interferon-α (IFN-α) and infectious virus shed after 10 focus-forming units (ffu) or 1000 ffu of human rotavirus (HRV). Diarrhoea scores (a), percentage of piglets with jejunal pathology (b), serum IFN-α (pg/ml, c) and infectious virus shedding (ffu/ml, d) are shown after 10 ffu (○) and 1000 ffu (▪) from post-infection day (PID) 0–6 (

*x*-axis). The number of piglets studied and the percentage of piglets that developed diarrhoea, shedding and jejunal pathology at each time-point is shown in each graph. Data-points with an asterisk differ significantly between doses at each time-point. The error bars denote the standard error of the mean (SEM) and values with an asterisk denote significant differences between groups at each PID. The HRV faecal shedding and diarrhoea scores were analysed using analysis of variance followed by Duncan's rank sum test, the percentage of pigs with diarrhoea and RV shedding was analysed using Fisher's exact test. Serum IFN-α was analysed using Kruskall–Wallis rank sum test. A*P*< 0.05 was considered the limit of statistical significance.7.

Total frequencies of splenic and mesenteric lymph node (MLN) plasmacytoid dendritic cells (pDCs), conventional dendritic cells (cDCs) and macrophage/monocytes after human rotavirus (HRV) infection. (a) spleen and (e) MLN, representative dot plots of SWC3 versus CD4 at post-infection day (PID) 2, 4 and 7 after HRV inoculation (left graph) or inoculation with medium alone (right graph) are shown. Gates and percentages of SWC3

^{low}CD4^{+}DCs (pDCs), SWC3^{high}CD4^{−}DCs (macrophage/monocytes) and SWC3^{low}CD4^{−}DCs (cDCs) are shown. Mean total frequencies of splenic pDCs (b), macrophages/monocytes (c) and cDCs (d) or MLN pDCs (f), macrophages/monocytes (g) and cDCs (h) in piglets that received HRV (○) or media alone (▪) are summarized. For each experiment 100 000 events were acquired by the flow cytometer. The number of piglets studied at each PID is shown at the bottom of each graph. The error bars denote the standard error of the mean (SEM) and time-points with an asterisk differ significantly from the controls at each PID (Kruskall-Wallis rank sum test*P*≤ 0.05).