A 27 kDa Cysteine Protease Secreted by Newly Excysted Paragonimus westermani Metacercariae Induces Superoxide Anion Production and Degranulation of Human Eosinophils

  • Chung, Young-Bae (Department of Parasitology, Cheju National University College of Medicine) ;
  • Kita, Hirohito (Department of Internal Medicine and Immunology, Mayo Clinic and Mayo Foundation) ;
  • Shin, Myeong-Heon (Department of Environmental Medical Biology, Institute of Tropical Medicine, and Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine)
  • 발행 : 2008.06.30

초록

Eosinophil degranulation plays a crucial role in tissue inflammatory reactions associated with helminth parasitic infections and allergic diseases. Paragonimus westermani, a lung fluke causing human paragonimiasis, secretes a large amount of cysteine proteases, which are involved in nutrient uptake, tissue invasion, and modulation of hos's immune responses. There is, however, limited information about the response of eosinophils to direct stimulation by cysteine proteases (CP) secreted by P. westermani. In the present study, we tested whether degranulation and superoxide production from human eosinophils can be induced by stimulation of the 2 CP (27 kDa and 28 kDa) purified from excretory-secretory products (ESP) of P. westermani newly excysted metacercariae (PwNEM). A large quantity of eosinophil-derived neurotoxin (EDN) was detected in the culture supernatant when human eosinophils isolated from the peripheral blood were incubated with the purified 27 kDa CP. Furthermore, the 27 kDa CP induced superoxide anion production by eosinophils in time- and dose-dependent manners. In contrast, the purified 28 kDa CP did not induce superoxide production and degranulation. These findings suggest that the 27 kDa CP secreted by PwNEM induces superoxide production and degranulation of human eosinophils, which may be involved in eosinophil-mediated tissue inflammatory responses during the larval migration in human paragonimiasis.

키워드

참고문헌

  1. Chung YB, Kong Y, Joo IJ, Cho SY, Kang SY. Excystment of Paragonimus westermani metacercariae by endogenous cysteine protease. J Parasitol 1995; 81: 137-142. https://doi.org/10.2307/3283911
  2. Na BK, Kim SH, Lee EG, Kim TS, Bae YA, Kang I, Yu JR, Sohn WM, Cho SY, Kong Y. Critical roles for excretory-secretory cysteine proteases during tissue invasion of Paragonimus westermani newly excysted metacercariae. Cell Microbiol 2006; 8: 1034-1046. https://doi.org/10.1111/j.1462-5822.2006.00685.x
  3. Shin MH, Kita H, Park HY, Seoh JY. Cysteine protease secreted by Paragonimus westermani attenuates effector functions of human eosinophils stimulated with immunoglobulin G. Infect Immun 2001; 69: 1599-1604. https://doi.org/10.1128/IAI.69.3.1599-1604.2001
  4. Shin MH, Lee SY. Proteolytic activity of cysteine protease in excretory- secretory product of Paragonimus westermani newly excysted metacercariae pivotally regulates IL-8 production of human eosinophils. Parasite Immunol 2000; 22: 529-533. https://doi.org/10.1046/j.1365-3024.2000.00337.x
  5. Shin MH, Seoh JY, Park HY, Kita H. Excretory-secretory products by Paragonimus westermani delay spontaneous cell death of human eosinophils through autocrine production of GM-CSF. Int Arch Allergy Immunol 2003; 132: 48-57. https://doi.org/10.1159/000073264
  6. Gleich GJ, Adolphson CR. The eosinophilic leukocytes: structure and function. Adv Immunol 1986; 39: 177-253. https://doi.org/10.1016/S0065-2776(08)60351-X
  7. Horie S, Gleich GJ, Kita H. Cytokines directly induce degranulation and superoxide production from human eosinophils. J Allergy Clin Immunol 1996; 98: 371-381. https://doi.org/10.1016/S0091-6749(96)70161-6
  8. Shin MH, Min DY. Infection status of Paragonimus westermani metacercariae in crayfish (Cambaroides similis) collected from Bogildo (Islet), Wando-gun, Chollanam-do, Korea. Korean J Parasitol 1999; 37: 55-57. https://doi.org/10.3347/kjp.1999.37.1.55
  9. Shin MH, Lee YA, Bae YS, Kita H, Kim Y, Ryu SH. The synthetic chemoattractant peptide WKYMVm induces superoxide production by human eosinophils via the phosphoinositide 3-kinase mediated activation of ERK1/2. Int Arch Allergy Immunol 2005; 137: S21-S26. https://doi.org/10.1159/000085428
  10. Shin MH, Chung YB, Kita H. Degranulation of human eosinophils induced by Paragonimus westermani-secreted protease. Korean J Parasitol 2005; 43: 33-37. https://doi.org/10.3347/kjp.2005.43.1.33
  11. Lee JC, Cho GS, Kwon JH, Shin MH, Lim JH, Kim WK. Macrophageal/ microglial cell activation and cerebral injury induced by excretory-secretory products secreted by Paragonimus westermani. Neurosci Res 2006; 54: 133-139. https://doi.org/10.1016/j.neures.2005.11.001
  12. Jin Y, Lee JC, Choi IY, Kim EA, Shin MH, Kim WK. Excretorysecretory products produced by Paragonimus westermani differentially regulate the nitric oxide production and viability of microglial cells. Int Ach Allergy Immunol 2006; 139: 16-24. https://doi.org/10.1159/000089518
  13. Carmona C, Dowd AJ, Smith AM, Dalton JP. Cathepsin L proteinases secreted by Fasciola hepatica in vitro prevent antibodymediated eosinophil attachment to newly excysted juveniles. Mol Biochem Parasitol 1993; 62: 9-17. https://doi.org/10.1016/0166-6851(93)90172-T
  14. Kong Y, Chung YB, Cho SY, Kang SY. Cleavage of immunoglobulin G by excretory-secretory cathepsin S-like protease of Spirometra mansoni plerocercoid. Parasitology 1994; 109: 611-621. https://doi.org/10.1017/S0031182000076496
  15. Barrett AJ, Kirschke H. Cathepsin B, cathepsin H, and cathepsin L. Methods Enzymol 1981; 80: 535-561. https://doi.org/10.1016/S0076-6879(81)80043-2
  16. Shin MH. Excretory-secretory product of newly excysted metacercariae of Paragonimus westermani directly induces eosinophil apoptosis. Korean J Parasitol 2000; 38: 17-23. https://doi.org/10.3347/kjp.2000.38.1.17
  17. Min DY, Lee YA, Ryu JS, Ahn MH, Chung YB, Sim S, Shin MH. Caspase-3-mediated apoptosis of human eosinophils by the tissue- invading helminth Paragonimus westermani. Int Arch Allergy Immunol 2004; 133: 357-364. https://doi.org/10.1159/000077355
  18. He S, Walls AF. Human mast cell chymase induces the accumulation of neutrophils, eosinophils, and other inflammatory cells in vivo. Br J Pharmacol 1998; 125: 1491-1500. https://doi.org/10.1038/sj.bjp.0702223
  19. Liu H, Lazarus SC, Caughey GH, Fahy JV. Neutrophil elastase and elastase-rich cystic fibrosis sputum degranulate human eosinophils in vitro. Am J Physiol 1999; 276: L28-L34.
  20. Temkin V, Kantor B, Weg V, Hartman ML, Levi-Schaffer F. Tryptase activates the mitogen-activated protein kinase/activator protein- 1 pathway in human peripheral blood eosinophils, causing cytokine production and release. J Immunol 2000; 169: 2662- 2669.
  21. Lee EG, Na BK, Bae YA, Kim SH, Je EY, Ju JW, Cho SH, Kim TS, Kang SY, Cho SY, Kong Y. Identification of immunodominant excretory-secretory cysteine proteases of adult Paragonimus westermani by proteome analysis. Proteomics 2006; 6: 1290-1300. https://doi.org/10.1002/pmic.200500399