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. 2016 Jul:36:305-314.
doi: 10.1016/j.intimp.2016.04.036. Epub 2016 May 21.

Chicken egg yolk antibodies (IgY) modulate the intestinal mucosal immune response in a mouse model of Salmonella typhimurium infection

Xiaoyu Li  1 , Ying Yao  2 , Xitao Wang  2 , Yuhong Zhen  3 , Philip A Thacker  4 , Lili Wang  1 , Ming Shi  5 , Junjun Zhao  6 , Ying Zong  2 , Ni Wang  2 , Yongping Xu  7
Affiliations

Chicken egg yolk antibodies (IgY) modulate the intestinal mucosal immune response in a mouse model of Salmonella typhimurium infection

Xiaoyu Li et al. Int Immunopharmacol. 2016 Jul.

Abstract

This study determined the effects of chicken egg yolk antibodies (IgY) on immune responses in the intestinal mucosal of mice infected with Salmonella typhimurium. Sixty, 28-day-old mice were divided into 4 groups and treated with streptomycin or sterile water for 2days followed by 1day without treatment. The control group was unchallenged whereas the mice in the other three groups were treated twice with 10(9)CFUmL(-1)S. typhimurium. For the next 3days, control mice continued to receive no treatment whereas the mice in the remaining three groups were orally administered with 20mgmL(-1) of specific IgY, 20mgmL(-1) of nonspecific IgY or PBS. S. typhimurium activated gut-associated lymphoid tissue, increasing the release of IFN-γ and TNF-α in the mucosa and increased the number of activated T-lymphocytes and cytotoxic T-γδ. Specific IgY attenuated the increase in IFN-γ and TNF-α and the decrease in IL-10. S. typhimurium induced mobilization of CD8(+) and CD8(+) TCRγδ T cells in the epithelium and CD4(+) and CD8(+) T cells in the lamina propria reflecting an inflammatory process that was attenuated by IgY. These results suggest that specific IgY modulates intestinal mucosal immune responses during a S. typhimurium infection.

Keywords: Egg yolk antibodies; Gut-associated lymphoid tissue; IgY; Intestinal inflammation; Mucosal immune response; Salmonella typhimurium.

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Figures

Fig. 1
Fig. 1
A. Survival rate of mice orally challenged with 0.4 mL of viable S. typhimurium organisms (109 CFU mL− 1 per mouse) at 0 and 24 h. Unchallenged mice received no treatment, whereas the challenged mice were treated once a day (after the 2nd bacterial challenge) for 7 consecutive days with 0.4 mL of 20 mg mL− 1 specific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS (n = 15 per group). Data were analyzed by the Chi-square test. *P < 0.05 compared with no treatment. B. Weight gain of unchallenged mice receiving no treatment, untreated mice challenged with S. typhimurium and challenged mice treated with 0.4 mL of 20 mg mL− 1 nonspecific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS 7 days post-challenge (n = 6 per group). Data are expressed as means ± SD and analyzed by ANOVA followed by SNK. Means without the same letter are significantly different (P < 0.05).
Fig. 2
Fig. 2
Representative histopathology of the jejunum from unchallenged mice receiving no treatment, untreated mice challenged with S. typhimurium and challenged mice treated with 0.4 mL of 20 mg mL− 1 nonspecific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS 7 days post-challenge. (A, C, E, G) Magnification: 200 ×. Jejunum mucosa sections show a focal area of rupture and ulceration after challenge that was cured after administration of IgY (arrows). (B, D, F, H) Magnification: 400 ×. The mucosa, submucosa and remaining jejunum wall show evidence of inflammation after challenge which was minimized by administration of IgY (arrows).
Fig. 3
Fig. 3
Pathological score analysis for HE staining. Edema in the submucosa (A), leucocyte infiltrates (B) and epithelial damage (C) was scored separately. The combined score equals the sum of the separate scores (D). Statistical analyses are shown for the separate scores and for the combined score. Data are expressed as means ± SD and analyzed by ANOVA followed by SNK (n = 6 per group). Means without the same letter are significantly different (P < 0.05).
Fig. 4
Fig. 4
mRNA levels of TNF-α (A), IFN-γ (B) and IL-10 (C) in the jejunal mucosal from unchallenged mice receiving no treatment, untreated mice challenged with S. typhimurium and challenged mice treated with 0.4 mL of 20 mg mL− 1 nonspecific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS 7 days post-challenge (n = 6 per group). Data are expressed as means ± SD and analyzed by ANOVA followed by SNK. Means without the same letter are significantly different (P < 0.05).
Fig. 5
Fig. 5
Representative images of the immunohistochemical localization of lymphocyte populations in the jejunum of unchallenged mice receiving no treatment, untreated mice challenged with S. typhimurium and challenged mice treated with 0.4 mL of 20 mg mL− 1 nonspecific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS 7 days post-challenge (n = 6 per group). Jejunum sections immuno-stained with anti-CD3, anti-CD4, anti-CD8, anti-TCRγδ and counterstained with hematoxylin (nuclear marker) are shown at 40 × magnification. Staining for lymphocyte markers is shown in brown and nuclei in blue.
Fig. 6
Fig. 6
Quantification of lymphocyte populations in the jejunum of unchallenged mice receiving no treatment, untreated mice challenged with S. typhimurium and challenged mice treated with 0.4 mL of 20 mg mL− 1 nonspecific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS 7 days post-challenge (n = 6 per group). Values are means ± SD and analyzed by ANOVA followed by SNK. Means without the same letter are significantly different (P < 0.05).
Fig. 7
Fig. 7
Representative plots of lymphocyte subsets in the small intestine of unchallenged mice receiving no treatment, untreated mice challenged with S. typhimurium and challenged mice treated with 0.4 mL of 20 mg mL− 1 nonspecific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS 7 days post-challenge (n = 6 per group). Lymphocytes from the intestinal intra-epithelium, lamina propria and Peyer's patches were isolated from the small intestine as described, pooled, and stained with different antibodies to identify various immune cells (CD3, CD4, CD8, CD25, and TCRγδ). Values are presented as the percentage of total lymphocytes. Figures shown are representative of three individual experiments.
Fig. 8
Fig. 8
Effects of IgY on lymphocyte populations in the intestinal intra-epithelium (A), lamina propria (B) and Peyer's patches (C) of unchallenged mice receiving no treatment, untreated mice challenged with S. typhimurium and challenged mice treated with 0.4 mL of 20 mg mL− 1 nonspecific IgY, 0.4 mL of 20 mg mL− 1 nonspecific IgY or 0.4 mL of PBS 7 days post-challenge (n = 6 per group). Values are means ± SD and analyzed by ANOVA followed by SNK. Means without the same letter are significantly different (P < 0.05).
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