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The effects of antibodies and complement in macrophage-mediated cytotoxicity on metacercariae of the lung fluke, Paragonimus westermani
D Y Min,M H Ahn,K M Kim,M H Leem and S Y Park
Department of Parasitology, College of Medicine, Hanyang University, Seoul 133-791, Korea.
Abstract
Paragonimus westermani is a tissue migrating parasite in the early stage until arriving at lung, and most of the parasites spend their life spans there. Considerable immune responses including activation of macrophages are taken place during the residence of parasites in the host. However, concerning the immunologic defense mechanisms of the host against this parasite, only a few document is available so far. In this study, the cytotoxic effect of peritoneal macrophages under the presence of antibody and/or complement against metacercariae of P. westermani was investigated in vitro.
Metacercariae were collected from the crayfish, Cambaroides similis and hatched out in Tyrode solution (pH 7.4). Plastic adherent cells from normal or infected rat (Wistar) peritoneal exudates were used as experimental macrophages. Polyclonal antibodies were obtained from infected rats and a cat. Cat IgG was fractioned with ion exchange chromatography. Fresh rabbit complement was used according to experimental scheme. Various combinations of peritoneal macrophages, normal or infected rat serum, complement and cat IgG were incubated at 36℃ in 5% CO2 incubator for 6, 14, 24 and 48 hours.
The results obtained were as follows:
1. P. westermani infection activated peritoneal macrophages non-specifically and this activation induced increases of cell adherence and cytotoxicity on metacercariae.
2. In the presence of infected rat serum the antibody-dependent cell-mediated cytotoxicity of peritoneal macrophages on metacercariae was significantly increased and showed a peak at 6-hour incubation. But the cytotoxic effect was markedly reduced after inactivation of complement and heat-labile IgE antibody by the heating of infected serum at 56℃ for 30 minutes.
3. The highest cytotoxic effect (100%) of concomitant incubation with IgG and complement showed 24 hours after incubation, although cell adherence was relatively low at 6-hour incubation and 0% at 24-hour incubation.
4. Coordinative functions of complement with serum and IgG were effective in cell adherence and in cytotoxicity, but it is not clear the independent role of complement on the macrophage-mediated cytotoxicity in this study.
With these results it is assumed that P. westermani infection can induce the non-specific activation of peritoneal macrophages, and serum antibodies including IgE antibody might enhance the cytotoxicity by macrophages.
Figures
Fig. 1 P. westermani metacercariae incubated with rat peritoneal macrophages in RPMI/FCS. After 6-hour incubation in various conditions cell adherence was classified as negative response (a), moderate (b) or extensive attachment (c).
Fig. 2 The peritoneal macrophage adherence on P. westermani metacercaria after incubation with various conditions.
(▵-▵: cells from normal rat, ▪-▪: cells from infected rat, ✕-✕: cells from normal rat + normal rat serum, •-•: cells from infected rat + normal rat serum).
Fig. 3 The effects of fresh and inactivated sera from P. westermani infected rat after incubation with normal peritoneal macrophages.
(•-•: normal rat serum, ▪-▪: infected rat serum, ◦-◦: heat-inactivated infected serum at 56℃ for 30 minutes).
Tables
Table 1 The percent of metacercariae showing peritoneal macrophage adherence after being incubated with cells, sera or IgG antibody from normal rats or rats infected with P. westermani
Table 2 The number of non-motile metacercariae after incubation with anti-serum or IgG antibody and peritoneal macrophages from normal rat
Table 3 Motility changes of survived metacercariae after incubation with antiserum or IgG antibody and peritoneal macrophages from normal rat
Table 4 The percent of metacercariae showing peritoneal macrophage adherence after being incubated with cells and sera from paragonimiasis patient
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