BackgroundRat mast cells were regarded as a good model for mast cell function in immune response.MethodsRat bone marrow mast cells (BMMC) were prepared both by recombinant rat IL-3 (rrIL-3) and by recombinant mouse stem cell factor (rmSCF), and investigated for both proliferation and differentiation in time course. Rat BMMC was induced by culture of rat bone marrow cells (BMCs) in the presence of both rrIL-3 (5 ng/ml) and rmSCF (5 ng/ml). Culture media were changed 2 times per week with the cell number condition of 5×104/ml in 6 well plate. Proliferation was analyzed by cell number and cell counting kit-8 (CCK-8) and differentiation was by rat mast cell protease (RMCP) II and histamine.ResultsCell proliferation rates reached a maximum at 8 or 11 days of culture and decreased thereafter. However, both RMCP II production and histamine synthesis peaked after 11 days of culture. By real time RT-PCR, the level of histidine decarboxylase mRNA was more than 500 times higher on culture day 11 than on culture day 5. By transmission electron microscopy, the cells were heterogeneous in size and contained cytoplasmic granules. Using gated flow cytometry, we showed that cultured BMCs expressed high levels of FcεRI and the mast cell antigen, ganglioside, on culture day 11.ConclusionThese results indicate that rat BMMCs were generated by culturing BMCs in the presence of rrIL-3 and rmSCF and that the BMMCs have the characteristics of mucosal mast cells.

Abstract:The third-stage larvae (L3) of the parasitic nematode, Anisakis simplex, have been implicated in the induction of hyperimmune allergic reactions in orally infected humans. In this work, we have conducted a review of an investigation into immune reactions occurring in animals experimentally infected with A. simplex L3. The patterns of serum antibody productions in the experimental animals against excretory-secretory products (ESP) of A. simplex L3 contributed to our current knowledge regarding specific humoral immune reactions in humans. In our review, we were able to determine that L3 infection of experimental animals may constitute a good model system for further exploration of immune mechanisms and allergy in anisakiasis of humans. worldwide (Mattiucci et al., 1997;Shih, 2004;Ugland et al., 2004). Many epidemics have been reported in Japan and Spain (Chai et al., 2005). Increasing reports of L3 infection in fish have been documented in South Korea, in which the consumption of raw marine fish is also popular (Chai et al., 1992;Im et al., 1995;Song et al., 1995Song et al., , 1999. Easy access to endoscopes and enhanced awareness of anisakiasis among clinicians has resulted in better reporting of morbidity resultant from L3 infection. L3 infection induces the production of specific antibodies and cytokines (Kennedy et al., 1988;Daschner et al., 2001;Nieuwenhuizen et al., 2006). Antibodies can be detected 2 weeks after infection, consistent with the time courses associated with other microorganisms. Analyses of specific antibody levels are generally irrelevant to the differential diagnosis of an acute state in cases of L3 infection, because the profound pain associated with L3 penetration begins only a few hours after the consumption of infected raw fish. However, antibody level measurements are helpful both in the differentiation of tumors from the granulomas formed by infiltrating L3 and in investigations of allergic diseases (Gutierrez and Cuellar, 2002;Kim et al., 2006).The production of IgE tends to increase during parasite infections, but the ultimate effects of IgE vary considerably, depending on the host-parasite relationships. Hyperimmune allergic reactions have been closely associated with IgE production. The infection of a parasite into its normal host tends to reduce the development of allergic responses, despite the associated upshift in IgE production (van den Biggelaar et al., 2000;Yazdanbakhsh et al., 2002). By contrast, the infection of T. canis in humans, an abnormal host, induces increased allergic reactions (Sharp and Olson, 1962;Sharghi et al., 2001). A. simplex L3 has also been shown to induce allergic diseases at a high rate, principally due to the fact that humans are not a regular host of this parasite (Audicana et al., 2002;Klimpel et al., 2004).Through investigations of allergic responses to L3 for a period of more than 10 years, several shared features have been identified, which indicate that immune reactions to L3 infection evidence similar patterns in humans and experimental...