Helminth infections in mammals are characterized by a high level of eosinophils in parasitized tissues and blood, and it has recently been suggested that these cells have a direct parasiticidal effect. Newborn larvae of Trichinella spiralis can be killed within 20 min by incubation at room temperature in a cell-free system, including purified human eosinophil peroxidase (EPO), H2O2 and chloride at pH 5.5. Killing was measured by microscopic observation of the larvae. The larvicidal effect was dependent on each component of the EPO/H2O2/Cl- system and could be prevented by using SO4(2-) instead of Cl-. Killing was totally inhibited by sodium azide and catalase, and substantially by bovine serum albumin, a protein that is an effective scavenger for HOCl. Since larvae could also be killed directly by HOCl under these conditions and EPO is able to oxidize Cl- to hypochlorous acid, it is very likely that the larvicidal effect of the EPO system is due to formation of hypochlorous acid. It is proposed that in vivo, the combination of EPO, which is exocytosed onto the surface of the parasite, and H2O2, which is generated by stimulated eosinophils, is responsible for the larvicidal effect.
The anti-parasite response was investigated after oral infection of athymic nude (rnu/rnu) rats and heterozygous (+/rnu) littermates with 1000 muscle larvae of Trichinella spiralis. No IgM, IgG and IgE antibodies were detected in serum of rnu/rnu rats. Expulsion of adult worms from the small intestine was prolonged (worms were nearly all expelled at days 14 and 91 in +/rnu and rnu/rnu rats respectively). The yield of muscle larvae in the carcasses of nude rats at day 91 was 33 times higher than in +/rnu rats. In contrast to the strong inflammatory reaction in the parasitized tongue of +/rnu rats, no infiltration was observed in rnu/rnu rats. Using an immunoperoxide method with monoclonal anti-rat T-cell antibody, no T cells were identified in spleen, mesenteric lymph node and Peyer's patches. These data support earlier studies that the nude rat lacks functional T cells. As the counts of connective tissue mast cells (CTMC), intestinal mast cells (IMC) and globule leucocytes (GL) in small intestine of uninfected rnu/rnu rats were equal or higher than in +/rnu rats, it is concluded that the origin of these cells is thymus-independent. In contrast to +/rnu rats, infection of rnu/rnu rats induced no increase of CTMC, IMC or GL. Thus, these cells depend on T cells to undergo proliferation. Finally, results of this study were inconclusive whether IMC are precursors for GL, or that they represent independent cell populations.
After immunization of Wistar rats with different concentrations of ovalbumin (OA) emulsified in complete H37Ra adjuvant, the optimal antigen dose for specific IgM, IgG and IgE production and for delayed-type hypersensitivity appeared to be the same. It is therefore concluded that in the rat these function tests can be combined, which decreases the number of experimental animals needed. For the determination of IgM, IgG and IgE antibodies to OA, the enzyme-linked immunosorbent assay (ELISA) was used, which proved to be a specific method. In addition, using a multi-channel photometer, it is shown that specific antibody concentrations can be readily quantified by micro-ELISA instead of using end-point titrations.
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