Fates of degranulated mast cells and extruded granules responded to homologous antigen in rats infected with <i>Clonorchis sinensis</i>

, Ahn, Yung Kyum; , Soh, Chin Thack

doi:1977.15.2.93

Korean J Parasito > Volume 15(2):1977 > Article

Original Article


Korean J Parasitol. 1977 Dec;15(2):93-99. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1977.15.2.93
Copyright © 1977 by The Korean Society for Parasitology


Fates of degranulated mast cells and extruded granules responded to homologous antigen in rats infected with Clonorchis sinensis
Yung-Kyum Ahn and Chin-Thack Soh
Department of Parasitology, Yonsei University Medical College, and Institute of Tropical Medicine, Yonsei University, Korea.


Abstract
Fates of degranulated mast cells by challenge of the homologous antigen to the Clonorchis sinensis infected rats, and the extruded granules were examined by means of light and electron microscopy. The whole experiment was carried out six weeks after the Clonorchis infection and an observation was made at 1, 3, 5, 8, 12 hours after the antigen challenges. The abdomen of the infected rat was cut open by 2 centimeters and the antigen was directly inoculated to mesentery, then again sutured. According to the scheduled period, the rat was sacrificed. The challenged portion was cut out and followed by dying procedure. The results of the observation are summarized as follows: Degranulation was observed within one hour without noticeable change of the granules. Three hours after the inoculation, the shed granules were phagocytized by macrophage surrounding the mast cells. They were aggregated in the cytoplasm. In 5 days phagocytosis phenomenon were almost completed but still some granules were scattered in surroundings upto 12 hours. The nucleus of the degranulated cell appeared clearly in contrast to normal cells which were killed with granules. Membrane became to normal, and the granules which were not expelled out agglomerated in a large cavity. The above resulst suggest that the partially degranulated mast cells do not disintegrate, but recover to normal, and expelled granules are phagocytized by macrophages.

Figures

Figs. 1-3
Light micrographs of the mesenteric mast cells from rats infected with Clonorchis sinesis. Stained with toluidine blue.

Fig. 1. Mesenteric mast cells and macrophages of rats of nonchallenged (×10)

Fig. 2. Degranulated mast cells and the extruded granules. Many granules are phagocytized by macrophages(arrows). a. after challenge with homologous antigen for 3 hrs (×400). b. higher magnification of figure 2a(×1,000). c,d and e. after challenge for 5 hrs, 8 hrs and 12 hrs(×400)

Fig. 3.a. The mast cells recovered from degranulation after 24 hrs (×400). b. higher magnification (×1,000). The large nucleus in the cytoplasm can be observed.

Figs. 4-7
Fig. 4. Electron micrograph of a normal mesenteric mast cell from non-challenged rat. The cytoplasm in tightly packed with granules having a uniformly electron dense, osmiophilic granules(×10,000). A few mitochondria (Mi) and endoplasmic reticulum (Er) are scattered in the cytoplasm. A small number of nuclear pores (Np) are evident in the section. Several microvilli(Vi) extend from the cell surface.

Fig. 5. A mast cell challenged with homologous antigen for 30 seconds. The extruded granules are seen with swollen appearance and weak electron dense, intact granules in the cell remained unchanged(×10,000) eG : extruded granule, sG : swollen granule, Ac : anateomosing cavity.

Fig. 6. Macrophage(Mac) is engulfing a freed granule (eG).

Fig. 7. Recovered phase of mast cell after 24 hours of the challenge (×18,000). The swollen granules remain in the cell combining with each other in a large in a large cavity (Lc, small arrows). Membrane closed to the expelled granule became nomal (large arrow).

References


1.	Chi EY, Lagunoff D, Koehler JK. Electron microscopy of freeze-fractured rat peritoneal mast cells. J Ultrastruct Res 1975;51(1):46–54.

2.	Combs JW. Maturation of rat mast cells. An electron microscope study. J Cell Biol 1966;31(3):563–575.

3.	Higginbotham RD, Dougherty TF, Jee WS. Fate of shed mast cell granules. Proc Soc Exp Biol Med 1956;92(2):256–261.

4.	Kagayama M, Douglas WW. Electron microscope evidence of calcium-induced exocytosis in mast cells treated with 48-80 or the ionophores A-23187 and X-537A. J Cell Biol 1974;62(2):519–526.

5.	Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.

6.	Mann PR. An electron microscope study of the degranulation of rat peritoneal mast cells brought about by four different agents. Br J Dermatol 1969;81(12):926–936.

7.	Nosal R, Slorach SA, Uvnas B. Quantitative correlation between degranulation and histamine release following exposure of rat mast cells to compound 48-80 in vitro. Acta Physiol Scand 1970;80(2):215–221.

8.	Rohlich P, Anderson P, Uvnas B. Electron microscope observations on compounds 48-80-induced degranulation in rat mast cells. Evidence for sequential exocytosis of storage granules. J Cell Biol 1971;51(21):465–483.