Characterization of isolated porcine intestinal mucosal mast cells following infection with Ascaris suum

Porcine intestinal mucosal mast cells (IMMC) were isolated from intestinal tissues of swine by enzymatic digestion and density gradient separation. Helminth-free swine and swine exposed to the nematode parasite, Ascaris suum, were used as a source of intestinal tissue. Up to 40% of the isolated intestinal cells stained metachromatically with toluidine blue pH 3.0, indicating the presence of IMMC. The histamine content of this cell population ranged from 2.9-8.9 pg per toluidine blue-positive IMMC, regardless of the animal source. Enrichment procedures that increased the proportion of toluidine blue-positive IMMC from the isolated intestinal cell population correlated with an increase in the amount of histamine detected in the cell population, indicating that toluidine blue-positive IMMC were the major source of histamine in this heterogeneous cell population. However, only cells isolated from the intestines of parasite-exposed swine released histamine in vitro after mixing with antigens derived from A. suum. Cells from the intestines of both helminth-free and parasite-exposed swine did not release histamine after mixing with a non-parasite hapten-protein molecule DNP-human serum albumin, but did release greater than 90% of their total histamine after lysis with Triton X-100 or with the Ca2+ ionophore (A23187). The stimulus for acquired responsiveness of IMMC to A. suum antigens in vitro was parasitic infection in vivo because helminth-free swine maintained in confinement on concrete yielded IMMC that specifically released histamine in the presence of parasite antigens only after 3 weeks of daily experimental inoculations with A. suum eggs. IMMC isolated from the entire length of the small intestines of infected pigs were responsive to antigens in vitro, but the relative number of IMMC isolated and their level of histamine release decreased from the anterior to the posterior end. IMMC isolated from infected swine were also stimulated to release histamine in vitro by viable second stage larvae of A. suum and by treatment with anti-swine immunoglobulin. Responsiveness to both parasite antigens and anti-immunoglobulin were totally eliminated, however, by a brief treatment of the cells with acidic buffer, suggesting that an acid-dissociable cell-bound antibody molecule was responsible for specific antigen-induced histamine release by IMMC.