Uptake of ferritin by M cells in follicle-associated epithelium at various sites in the small and large intestines was examined in 4 healthy 5-week-old pigs by use of electron microscopy. A 2.5% solution of ferritin in saline was injected into ligated loops of the jejunum and ileum containing aggregations of lymphoid follicles (Peyer's patches), as well as into intestinal loops containing lymphoglandular complexes at the ileocecal junction, in the central colonic flexure, and in the rectum. As negative control, saline solution was injected into loops at identical localizations. After an exposure period of 2 hours, uptake of ferritin by M cells, but not by enteroabsorptive cells of the small and large intestines, was observed. Numbers of M cells with ferritin and total M cells were counted and the percentage was calculated. Total number of M cells was highest in lymphoglandular complexes in the rectum and lowest on domes of the ileal Peyer's patch. High numbers of M cells with ferritin were found on domes of the jejunal Peyer's patch, and in lymphoglandular complexes at the ileoceral entrance and in the rectum. Only a few M cells on domes of the ileal Peyer's patch and in lymphoglandular complexes in the central colonic flexure contained ferritin. The percentage of M cells with internalized ferritin was similar on domes of the ileal Peyer's patch, and in lymphoglandular complexes at the ileocecal junction and in the rectum. It was higher on domes of the jejunal Peyer's patches and lower in lymphoglandular complexes of the central colonic flexure. Ferritin was found in the apical tubulovesicular system, multivesicular bodies, and a few vacuoles in the central area of M cells. Ferritin was exocytosed into the lateral intercellular spaces next to M cells. Uptake of ferritin by intraepithelial cells in the follicle-associated epithelium could not be documented, but ferritin was present in vesicles of subepithelial macrophages.

Objective-To develop a system for analysis of immune response variables in the lymph draining the lung and to establish baseline data for clinically normal calves. Design-Surgery was performed on 6 calves to insert a cannula into the efferent lymphatic duct of the caudal mediastinal lymph node to create a long-term thoracic lymph fistula draining to the exterior. Lymph was collected daily, and on the fifth postoperative day, calves were exposed to an aerosol of cell culture medium (mock infection). For the next 10 days, lymph was collected for analysis and, on the tenth day, necropsy was performed. Animals-Six 6- to 8-week-old Holstein bull calves. Procedure-Daily lymph samples were evaluated for: flow rate; total and differential cell counts; and IgG, IgM, IgA, IgE, and protein concentrations. On days -4, -1, 1, 4, 7, and 10, cells were stained and quantitated by fluorescence-activated cell sorter analysis for T, B, CD4+, and CD8+ cells. Blood lymphocytes were evaluated on days -1 and 10 for comparison. Results-Flow was established for up to 25 days, with a mean rate between 11 and 22 ml/h. Protein concentrations in lymph and plasma did not indicate a protein drain. Although mean lymphocyte counts reflected a slight gradual decrease in lymph lymphocytes, this effect was not apparent in every calf, nor was the effect seen in blood lymphocytes. There were no significant changes in IgG, IgM, IgA, or IgE concentration, with the exception of IgA concentration in 1 calf that developed an abscess at the cannulation site. The T-cell subset absolute numbers of CD4+ and CD8+ cells decreased slightly over time, but the CD4+-to-CD8+ cell ratio remained almost constant at near 2. Conclusion-Creation of a thoracic lymphatic fistula appears to be a useful technique for studying effects of lung infection on immunologic variables, with potential application to bacterial and viral respiratory tract diseases.