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Korean J Parasitol > Volume 29(1):1991 > Article

Original Article
Korean J Parasitol. 1991 Mar;29(1):43-54. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1991.29.1.43
Copyright © 1991 by The Korean Society for Parasitology
Antibody-dependent rat macrophage-mediated damage to the excysted metacercariae of Paragonimus westermani in vitro
P R Chung,J K Chang and C T Soh*
Department of Parasitology, College of Medicine, Yonsei University, Seoul, Korea.

An in vitro immune effector mechanism against the target excysted metacercariae of Paragonimus westermani was demonstrated in the rat system. Peritoneal exudate cells, mainly macrophages from normal rats, showed adherence to and killing of excysted metacercariae of P. westermani in the presence of complement-independent serum from rats infected with Paragonimus metacercariae. These reactions were specific for the excysted metacercariae, as tissue-migrating juvenile worms were not affected.

Damage of excysted metacercariae of P. westermani due to antibody and macrophages was assessed by morphological observation, by cell adherence reaction and by the use of vital dyes. Trypan blue dye exclusion proved to be a reliable indicator of judging metacercarial viability. Electron microscopic studies demonstrated that macrophages reacted with fuzzy material on the tegumental surface and fine structures in the syncytium of the parasites. The tubular tunnels formed between the basement membrane and muscle layers of the damaged parasites were also noticeable. The relevance of these findings to cellular immunity in the early paragonimiasis was discussed.


Fig. 1
Antibody-dependent adherence of rat peritoneal macrophages to the excysted metacercariae of P. westermani. (A) in the presence of macrophages and sera from uninfected normal rats, the excysted metacercariae appear normal and cells do not adhere to worms; (B) I the presence of sera from infected rat and macrophages, cells adhere to the surface of worms; (C) blebbing of the tegument with strong cell adherence reaction appears on the surface of dead worm, in the presence of sera from the rats 2 weeks after infection. (×100).

Figs. 2-4
Fig. 2. An excysted metacercaria of P. westermani after incubation in media containing sera from uninfected rats shows the normal, intact structural features such as tegumental ridges (TR), tegumental syncytiun (TS) containing mitochondria (Mi) and small ovoidal dense granules, circular muscles (CM), and longitudinal muscle (LM) layers just underneath the basement membrane (BM) of the tegumental syncytium (×12,500).

Fig. 3. Appearance of an excysted metacercaria after 2-hr incubation in media containing macrophages (Mac) from uninfected and sera from infected rats. Note deposition of electron-dense fuzzy material (FM) onto the tegument. Syncytial cytoplasm contains spines (Sp) and intact dense granules (arrow heads) (×10,000).

Fig. 4. Antibody-dependent adherence of macrophages and damage to the excysted worm. The excysted metacercariae were incubated for 18 hrs in the presence of macrophages (Mac) from uninfected rats and serum from infected rats. Note macrophages with a great number of pseudopodia (Ps) onto the tegumental surface and dense granules surrounded by small vesicles in the syncytial cytoplasm (arrow heads). No tegumental ridges were observed (×7,500).

Figs. 5-6
Fig. 5. An excysted metacercaria and a macrophage incubated in the same effector system as in Fig. 4. The increased amount of fuzzy material (FM) between macrophage and tegumental surface of worm, and phagosome-like vacuoles (PV) in the macrophage were remarkable. Dense granules denatured in the small vesicles were also enlarged (×12,500).

Fig. 6. A dead worm incubated form 18 hrs in the presence of homologous macrophages and serum obtained from rats 2 weeks after infection. Tubular tunnels (Tt; arrow heads) formed between the basement membrane and muscle layers of a damaged parasite were noticeable. The fine structures in the tegumental syncytium were almost destroyed (×10,000).


Table 1
Mean worm recoveries from five rats infected with 50 Paragonimus metacercariae per animal*

Table 2
Yield of rat peritoneal macrophages (Mø) according to stimulators employed

Table 3
Cell adherence and death of excysted metacercariae of P. westermani after an 18-hour culture with immune sera collected from rats at one-week intervals*

Table 4
Cell adherence and viability of juvenile worms(=adolescariae) of P. westermani in various effector system*

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