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Title: Novel staining procedure to visualize the chicken Peyer’s patch in situ for expanded immunological research of this tissue in poultry

Author
item Holt, Peter
item Vaughn, Lara
item Moore, Randle
item Gast, Richard
item KAISER, PETE - INST OF ANIMAL HEALTH

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 5/10/2007
Publication Date: 7/9/2007
Citation: Holt, P.S., Vaughn, L.E., Moore, R.W., Gast, R.K., Kaiser, P. 2007. Novel staining procedure to visualize the chicken Peyer’s patch in situ for expanded immunological research of this tissue in poultry. Meeting Abstract.

Interpretive Summary:

Technical Abstract: Background: Poultry research on the role of the chicken Peyer’s patch (Pp) in directing the gut mucosal immune response has been limited due to the inability to readily visualize the Pp in unfixed tissue. We set about to develop a method to rapidly visualize the Pp in the chicken and then used this methodology to begin studying the immune capacity of this tissue. Methods: Different stains were tested for their ability to enhance detection of the Pp. Once a suitable staining process was found, a study was conducted to compare cytokine mRNA expression in Pp and cecal tonsils (CT), a second chicken gut lymphoid tissue, in noninfected and Salmonella enteritidis - infected hens. These two tissues were removed from noninfected and infected hens at various times post challenge and cytokine mRNA expression was examined using quantitative real time PCR (RT-PCR). Results: Intraluminal administration of the stain Eosin-Y followed by crystal violet enabled the rapid visualization of the Pp from the serosal surface. The Pp showed up as a whitish-pink ovoid while the surrounding tissue was a light purple. Light microscopy evaluation of fixed tissues revealed lymphoid tissue aggregations with multiple follicular units mirroring that observed for mammalian Pp. Little expression of interleukin (IL)-10 or IL-12 mRNA, in the presence or absence of infection, was observed in the Pp while mRNA expression of both cytokines was observed in CT. Expression of IL-1, 4, 6, 8, and interferon gamma mRNA was similar between the two tissues. Conclusions: Intraluminal Eosin-Y/crystal violet administration allows ready observation of chicken Pp. The Pp exhibit similar architecture to their mammalian counterparts. Different cytokine mRNA profiles are observed between Pp and CT indicating different cell populations and possibly different functions in these two gut lymphoid tissues.