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Background: For many years the central focus of research into gastrointestinal hypersensitivity reactions has been the mast cell population of the intestinal lamina propria. Since bile is known to deliver immunological mediators to the gastrointestinal tract, the possibility arises that extra-intestinal populations of mast cells may also contribute to IgE-mediated intestinal damage., Objectives: To characterize hepatic mast cells in the rat and to investigate the role of the hepatobiliary system in a model of IgE-mediated reactivity to dietary antigen., Methods: Wistar rats were passively sensitized with monoclonal antidinitrophenyl (DNP) IgE antibodies, and were later challenged orogastrically with DNP-HSA. Additional animals were sensitized, then bile duct-cannulated prior to antigen challenge. At various time points, liver and intestinal samples were collected for histological examination, and bile was collected and assayed for histamine and TNFalpha., Results: Hepatic mast cells display a mucosal mast cell-like phenotype, and are closely associated with the vessels of the portal triads. Orogastric antigen challenge led to a rapid and significant decline (P<0.0001) in detectable mast cells as a result of anaphylactic degranulation. The median number of granulated mast cells associated with each portal triad in liver sections declined from six per portal triad to one per portal triad post-antigen challenge. After 15 min, biliary histamine concentrations rose above background levels (P<0.01). TNFalpha was also detectable in the majority (4/6) of bile samples within 15 min of challenge. Histological examination of the gastrointestinal mucosa revealed disruption to the villous epithelium ranging from oedematous changes to gross destruction. Such damage was not seen in animals in which bile had been externally drained., Conclusion: The data indicate that biliary products are major contributors to the gastrointestinal damage arising from IgE-mediated hypersensitivity reactions in the rat, and such hypersensitivity reactions may involve a population of mast cells which reside in the liver.

The notion that bile-dedicated antibody is made within the liver by migratory antibody-forming cells (AFC) was examined further in rats. Livers from immunized animals were removed to perfusion in isolation so that plasma influences on bile antibody would be obviated. Antibody was secreted for at least 5 hr by the livers of rats that had received intravenous (i.v.) or intra-Peyer's patch (IPP) immunization with horse erythrocytes. After initially declining, the titres stabilized at 5-8% of the starting value for IPP-immunized rats and at 0.8% for i.v.-immunized animals, levels that were then sustained. In other experiments, the biliary antibody output was measured in immunized rats in the period immediately following splenectomy, an expedient that would deny the liver any newly formed AFC. Splenectomy during spleen-based, IgM antibody responses led to bile titres falling, over about 12 hr, to 21% of initial values. This level was then maintained for at least another 12 hr. Serum titres over this period remained static. Lastly, the bile ducts of immunized rats were ligated to test whether locally made antibody that was destined for bile could be forced instead to reflux to blood. Biliary obstruction during IgM responses to horse erythrocytes and pneumococcal polysaccharide, type 3, was found to raise significantly serum antibody titres. For pneumococcal polysaccharide, the serum response was also noticeably prolonged. These findings are consistent with the biliary antibody of immunized rats being constituted, in part, from local sources and not from plasma alone.

In the rat , ligation of the bile duct induces a rapid and progressive elevation of the IgA levels in serum. The increase is about 4-fold at 1 h, 15-fold at 1 day, and 30-fold at 1 wk after ligation. The additional IgA is of the secretory type. Free secretory component also appears in serum after bile duct obstruction; it does not continue to increase and occasionally disappears from serum after prolonged ligation. The increase in serum IgA levels is selective. These changes are totally reversible if the bile duct is reopened at 1 day after ligature. These findings confirm the role of the rat liver in the transfer of circulating IgA into the bile.

The transient appearance of IgM in the bile of rats injected intravenously with horse erythrocytes (HRBC) correlated with the occurrence of anti-HRBC IgM-secreting cells in the liver. Both responses were reduced in animals splenectomised at the time of immunisation. When rats were given 2 injections of HRBC spaced by 28 days, IgG, IgA and IgM antibody-forming cells (AFC) were detected in the liver along with IgM and non-IgM anti-HRBC antibody in bile. There was a 10-fold increase in the total numbers of mononuclear cells (MNC) retrievable from the liver at the height of the biliary antibody response, the majority of which were not AFC. These results suggest that antigen entering the spleen stimulates the release of a population of MNC which may localise in the liver and that a minor portion of these produces specific antibody which is secreted into bile.

Rat bile is an important source of s-IgA and FSC about 10 mg of s-IgA are poured daily in the duodenum via the bile. The normal rat liver is capable of actively transferring IgA from the circulation into bile against a strong concentration gradient. Circulating IgA transferred in bile was bound to SC. Ligature of the rat bile duct induces a large and selective increase of the IgA level in serum. All this additional IgA is s-IgA and even FSC appears in serum after bile duct obstruction. The liver appears to have an important reinforcing role in the intestinal s-IgA system of the rat.

The intravenous injection of 1 ml of a 0.1% (or higher %) suspension of horse red blood cells (HRBC) into rats results in the appearance of high levels of agglutinating activity in bile as well as serum. Lower doses of HRBC induce reduced responses in both fluids, demonstrating that the biliary response is as dose-dependent as the serum response. The presence of IgM anti-HRBC antibody forming cells (AFC) in the liver in numbers equivalent to those detected in the spleen at day 6 of the study suggests them to be the most likely source of the biliary antibody. Also, the injection of HRBC into other parenteral sites (intramuscular [i.m.], intraperitoneal [i.p.] and intrathoracic cavity [i.t.]) results in a biliary response which is considered to originate from the IgM-, IgG- and IgA-anti-HRBC AFC detected in the liver, again in numbers approximating those detected in the spleen. Splenectomy of rats immunized i.p., i.m. and i.t. had only a minimal effect on the liver and biliary responses, indicating that AFC arising in peripheral lymphoid tissues can also relocate in liver. It is concluded that the liver is a major site of antibody synthesis following parenteral immunization leading to substantial levels of antibody being secreted into the gastrointestinal tract.

Intestinal injection of HRBC into rats leads to: 1. an anti-HRBC response in serum and bile - HA activity in bile is associated with IgM, IgG and IgA. 2. an increase in total IgM in bile. 3. the presence of IgM, IgA and IgG AFC in liver. Equivalent AFC responses occur in livers of splenectomized rats. It is concluded that the results are consistent with the proposal that the liver is a site for localization of antibody-secreting cells which enter the blood and that these cells are a source of biliary antibody.