Absorption of IAIV by NALT on the septum.

<p>(A) Layer scanning by LSCM (×20), (B) lateral view of three-dimensional image (×20), (C) Merge of layer scanning images (×20), (D) Virtual HE stain of the merged image (×20). Essentially similar results were obtained in two independent experiments.</p

Absorption of 50 nm FLP by chicken nasal mucosa.

<p>(A) Stratified squamous epithelium (×5), (B) Mucosa of concha nasalis media (×16), (C) Mucous gland (×32), (D) NALTs (×32), (E) Backgroud control. The 50 nm FLP were fluorescent blue, the microfilament were staining with phalloidin (fluorescent green). Essentially similar results were obtained in two independent experiments.</p

Histological and anatomical structure of the nasal cavity of Bama minipigs

<div><p>Objective</p><p>The nasal mucosa is equipped with abundant lymphatic tissues, serving as the first line of defense against invasion by microorganisms. In this study, we characterized the features of the nasal mucosa of Bama minipigs (<i>Sus scrofa domestica</i>) via histological analysis.</p><p>Methods</p><p>Five cross sections (I, II, III, IV, and V) were obtained from the distal end of the nasal cavity toward the pharynx (along the cavity axis) and examined. Specifically, CD3⁺ T cells, immunoglobulin A (IgA)⁺ cells, and M cells were detected by immunohistochemistry, while dendritic cells (DCs) were detected by immunofluorescence. The distribution of goblet cells was determined by periodic acid-Schiff (PAS) staining.</p><p>Results</p><p>The nasal cavity of Bama minipigs can be divided into three parts: the regio vestibularis (I, II), regio respiratoria (III, IV), and regio olfactoria (V). Lymphoid tissue was present at random locations in the nasal cavity. Abundant lymphoid tissue was located in the roof of the nasopharyngeal meatus and was continuous with the lymphoid tissue of the pharynx. The distribution of CD3⁺ T cells, IgA⁺ cells, M cells, and DCs increased distally in the nasal cavity.</p><p>Conclusions</p><p>The present work comprises a histological study of the nasal cavity of Bama minipigs, and will be beneficial for understanding the mechanisms of immunity in these animals after nasal vaccination.</p></div

Changes of relative fluorescence area of IAIV in NALT.

<p>Significance of difference between groups (<i>P</i><0.01) was calculated using ANOVA. Identical upper-case letters indicate no statistical difference. Values are given as means ± SEM of 2 separate experiments.</p

BP5 enhanced the activities of antioxidant enzymes in the LPS-treated DCs.

<p>DCs (1×10⁶ cells/ml) were pretreated with BP5 for 2 h, followed by stimulation with or without LPS (100 ng/ml). After 22 h, Intracellular levels of (A) GPx, (B) CAT and (C) SOD were measured using commercial kits. Data shown are the means ± SD of three samples. *<i>P</i> < 0.05 in comparison with the LPS-only group; ^#<i>P</i> < 0.05 in comparison with the untreated group. All results are representative of three independent experiments.</p

Absorption of OVA by chicken nasal mucosa.

<p>(A) Medial side of the concha nasalis media, (B) Lateral side of the concha nasalis media, (C) Mucous gland, (D) NALT, (E) Backgroud control. Column I×100; Column II×200; Column III×400. Essentially similar results were obtained in three independent experiments.</p

BP5 suppressed lipid peroxidation in the LPS-stimulated DCs.

<p>DCs (1×10⁶ cells/ml) were pretreated with or without BP5 for 2 h, and then exposed to LPS (100 ng/ml) or not for 22 h. The MDA contents in cell lysate supernatants were measured as described in Materials and Methods. Data shown are the means ± SD of three samples. *<i>P</i> < 0.05 in comparison with the LPS-only group; ^#<i>P</i> < 0.05 in comparison with the untreated group. All results are representative of three independent experiments.</p

BP5 suppressed the DC maturation in the LPS-stimulated ECs/DCs coculture system.

<p>Experimental setting to study the DC maturation in the ECs/DCs coculture system. (A) The scheme depicts: Caco-2 cells were grown on the filter to form a tight monolayer, and then DCs were cultured facing the basolateral side of Caco-2 cells on the bottom of the filter for 4 h. Pretreatment of BP5 on the basolateral DCs for 2 h, medium or LPS (300 ng/ml) was incubated on the apical side of the ECs for 22 h, then the basolateral DCs and culture supernatants were collected. (B) TEER was measured by a Millicell-ERS epithelial voltohmmeter (Millipore) at indicated time. (C-D) The filters were fixed with 4% paraformaldehyde and then processed to immunofluorescence stain for CLSM. Three-dimensional rendering of representative fields was obtained with Imaris 7.2 software, DCs (red, CD11c), tight junction of ECs (green, occludin). Submucosal DCs sent dendrites (arrows) to creep through ECs in response to LPS but not medium. (E-F) Supernatants were collected and NO production was measured using the Griess reagent. (G-H) TNF-α released from basolateral supernatants was measured by ELISA. (F, H) DCs stimulated with LPS before, at the same time as, or after BP5 (100 μg/ml) incubation. (I-L) The expressions of CD86 and MHCII on DCs were analyzed by FACS. Data shown are the means ± SD of three samples. *<i>P</i> < 0.05; **<i>P</i> < 0.01. SN, supernatants. To confirm the results, we repeated these experiments three times.</p

BP5 upregulated the expression of HO-1 protein in the LPS-induced DCs.

<p>DCs (1×10⁶ cells/ml) were treated with the indicated concentrations of BP5 for 2 h, and then incubated with or without LPS (100 ng/ml) for 22 h. (A) HO-1 levels were assessed by western blotting. (B) Quantification of the blots. (C-H) DCs were pretreatment with BP5 (100 μg/ml) in the presence or absence of Snpp (25 μM) or Copp (50 μM) for 2 h, and then incubated with or without LPS (100 ng/ml) for 22 h. (C) Supernatants were collected and NO production was measured using the Griess reagent. (D) TNF-α released from supernatants was measured by ELISA. DCs were harvested and the expressions of CD80 (E, G) and CD86 (F, H) were analyzed by FACS. Data shown are the means ± SD of three samples. *<i>P</i> < 0.05; **<i>P</i> < 0.01. All results are representative of three independent experiments.</p