Cellular assessment of crop lymphoid tissue from specific-pathogen-free white leghorn chickens after Salmonella...

The crop may be an important site along the upper alimentary tract in which a humoral immune response against Salmonella Enteritidis (SE) is elicited locally. The mucosal immune response within the crop (ingluvies) of specific-pathogen-free (SPF) white leghorn (WL) chickens against SE was investigated. Three trials were conducted using SPF WL pullets at age 5-6 wk. Trial 1 consisted of 77 birds evaluated for 10 wk post-SE infection (pi), trial 2 was composed of 72 birds monitored through 8 wk pi, and trial 3 was made up of 30 birds assessed for 5 wk pi. Birds were challenged per os with 10(8) colony-forming units/ml SE phage type 13. Crop lavage samples, crop tissues, ceca, and/or liver-spleen were collected preinfection and then at weekly intervals post-SE infection. Bacteriologic examination of cecal contents and/or liver-spleen occurred weekly to monitor progression of SE infection. Crop lavages were analyzed for SE-lipopolysaccharide (LPS)-specific immunoglobulin A (IgA) by enzyme-linked immunosorbent assay to assess humoral immune response. General histologic staining (hematoxylin and eosin [H&E] and methyl green-pyronin [MGP]) and immunohistochemical (IHC) staining (monoclonal antibodies CD45 and Bu-1) were applied to serial sections of crop to evaluate lymphoid tissue via light microscopy, to grade isolated lymphoid follicles (ILFs) by using score 0 (minimal, < 50 microm in diameter) to score 5 (sizable, > 200 microm in diameter) scale, and to characterize the cellular population of ILFs. Results revealed that cecum samples and liver-spleen samples were 100% SE culture positive at 1 wk pi, and then the percentage of SE positives progressively declined over time. Markedly increased crop SE-LPS-specific IgA antibodies were detected in crop samples by 2-3 wk pi, and the humoral response remained elevated above week 0 baseline for the duration of each trial. Crop ILFs of score 3 to 5 were observed in H&E-stained tissues, with an increased proportion of ILFs in post-SE-infected crops vs. uninfected. MGP staining showed plasma cells scattered within and at the periphery of ILFs. IHC staining revealed CD45 (pan-leukocyte) and Bu-1 (B-lymphocyte)-positive cells within crop ILFs. The chicken crop seems to be an organ in which lymphoid tissue may arise in response to enteric SE infection, and a site in which a humoral response may be generated against the SE pathogen.