Journal Mobile Options
Table of Contents
Vol. 132, No. 1, 2003
Issue release date: September 2003
Int Arch Allergy Immunol 2003;132:48–57

Excretory-Secretory Products Secreted by Paragonimus westermani Delay the Spontaneous Cell Death of Human Eosinophils through Autocrine Production of GM-CSF

Shin M.H. · Seoh J.-Y. · Park H.-Y. · Kita H.
aDepartment of Parasitologyand Institute of Tropical Medicine, College of Medicine, Yonsei University, Departments of bMicrobiology and cBiochemistry and dMedical Research Center, College of Medicine, Ewha Womans University, Seoul, Korea; eDepartment of Immunology and Internal Medicine, Mayo Clinic and Mayo Foundation, Rochester, Minn., USA

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Background: Eosinophils play important roles in tissue inflammatory responses associated with helminth infections. Excretory-secretory products (ESP) produced by tissue-invasive helminths contain a large quantity of proteolytic enzymes that can modulate the host’s immune responses. However, little is known regarding the roles of worm-derived products that are responsible for eosinophilic inflammatory responses in helminth infections. Objective: In the present study, we investigated whether ESP produced by Paragonimus westermani, which cause pulmonary or extrapulmonary paragonimiasis in human beings, regulates both cell survival and death of human eosinophils. Methods: The ESP was obtained from P. westermani newly excysted metacercariae (PwNEM). Eosinophils were purified from peripheral blood of healthy donors, and the purified eosinophils were incubated with or without the ESP secreted by PwNEM. The viability of eosinophils was assessed by staining with propidium iodide using the flow cytometer. Results: When eosinophils were incubated with a low concentration of the ESP produced by PwNEM, which totally consists of proteolytic enzymes, eosinophil cell death was delayed compared with results for cells incubated with medium alone. In fact, the ESP at a low concentration stimulated eosinophils to produce detectable levels of GM-CSF that can delay eosinophil cell death. In contrast, eosinophil cell death was dose-dependently accelerated when cells were incubated with high concentrations of the ESP. To see whether the dose-dependent biphasic survival effect of the ESP on eosinophils is primarily due to the protease activity contained in the ESP, a high dose of the ESP was treated with heat at 56°C for 30 min before being added to eosinophils. Attenuating protease activity in a high dose of the ESP by heat treatment reversed the ESP-afforded eosinophil cell death. This prolonged survival of eosinophils induced by the heated ESP was remarkably inhibited by anti-GM-CSF-neutralizing mAb and Jak2 kinase inhibitor AG-490. Conclusion: These results suggest that the proteases in the ESP secreted by PwNEM are able to regulate eosinophil survival through the autocrine production of GM-CSF. Thus, the enhanced eosinophil survival induced by Paragonimus-secreted products may contribute to the elicitation of eosinophilic inflammatory responses at the worm-infected lesion in human paragonimiasis.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Gleich GJ, Adolphson CR: The eosinophilic leukocytes: Structure and function. Adv Immunol 1986;39:177–253.
  2. Gleich GJ: Mechanisms of eosinophil-associated inflammation. J Allergy Clin Immunol 2000;105:651–663.
  3. Kankaanranta H, Lindsay MA, Giembycz MA, Zhang X, Moilanen E, Barnes PI: Delayed eosinophil apoptosis in asthma. J Allergy Clin Immunol 2000;106:77–83.
  4. Ohta K, Yamashita N: Apoptosis of eosinophils and lymphocytes in allergic inflammation. J Allergy Clin Immunol 1999;104:14–21.
  5. Simon HU, Yousefi S, Schranz C, Schapowal A, Bachert C, Blaser K: Direct demonstration of delayed eosinophil apoptosis as a mechanism causing tissue eosinophilia. J Immunol 1997;158:3902–3908.
  6. Yousefi S, Blaser K, Simon HU: Activation of signaling pathways and prevention of apoptosis by cytokines in eosinophils. Int Arch Allergy Immunol 1997;112:9–12.
  7. Tai PC, Sun L, Spry CJ: Effects of IL-5, granulocyte/macrophage colony-stimulating factor (GM-CSF) and IL-3 on the survival of human blood eosinophils in vitro. Clin Exp Immunol 1991;85:312–326.
  8. Yamaguchi Y, Suda T, Ohta S, Tominaga K, Miura Y, Kasahara T: Analysis of the survival of mature human eosinophils: Interleukin-5 prevents apoptosis in mature human eosinophils. Blood 1991;78:2542–2547.
  9. Walsh GM, Symon FA, Wardlaw AJ: Human eosinophils preferentially survive on tissue fibronectin compared with plasma fibronectin. Clin Exp Allergy 1995;25:1128–1136.
  10. Kim JT, Gleich GJ, Kita H: Roles of CD9 molecules in survival and activation of human eosinophils. J Immunol 1997;159:926–933.
  11. Kim JT, Schimming AW, Kita H: Ligation of FcγRII (CD32) pivotally regulates survival of human eosinophils. J Immunol 1999;162:4253–4259.
  12. Levi-Schaffer F, Temkin V, Malamud V, Feld S, Zilberman Y: Mast cells enhance eosinophil survival in vitro: Role of TNF-α and granulocyte-macrophage colony-stimulating factor. J Immunol 1998;160:5554–5562.
  13. Meerschaert J, Busse WW, Bertics PJ, Mosher DF: CD14+ cells are necessary for increased survival of eosinophils in response to lipopolysaccharide. Am J Respir Cell Mol Biol 2000;23:780–787.
  14. Aga E, Katschinski DM, van Zandbergen G, Laufs H, Hansen B, Muller K, Solbach W, Laskay T: Inhibition of the spontaneous apoptosis of neutrophil granulocytes by the intracellular parasite Leishmania major. J Immunol 2002;169:898–905.
  15. Channon JY, Miselis KA, Minns LA, Dutta C, Kasper LH: Toxoplasma gondii induces granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor secretion by human fibroblasts: Implications for neutrophil apoptosis. Infect Immun 2002;70:6048–6057.
  16. Gon S, Saito S, Takeda Y, Miyata H, Takatsu K, Sendo F: Apoptosis and in vivo distribution and clearance of eosinophils in normal and Trichinella spiralis-infected rats. J Leukoc Biol 1997;62:309–317.
  17. Sugaya H, Abe T, Yoshimura K: Eosinophils in the cerebrospinal fluid of mice infected with Angiostrongylus cantonensis are resistant to apoptosis. Int J Parasitol 2001;31:1649–1658.
  18. Rumbley CA, Sugaya H, Zekavat SA, Perrin PJ, Phillips SM: Elimination of lymphocytes, but not eosinophils, by Fas-mediated apoptosis in murine schistosomiasis. Am J Trop Med Hyg 2001;65:442–449.
  19. Berasain P, Carmona C, Frangione B, Dalton JP, Goni F: Fasciola hepatica: Parasite-secreted proteinases degrade all human IgG subclasses: Determination of the specific cleavage sites and identification of the immunoglobulin fragments produced. Exp Parasitol 2000;94:99–110.
  20. Carmona C, Dowd AJ, Smith AM, Dalton JP: Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles. Mol Biochem Parasitol 1993;62:9–17.
  21. 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.
  22. 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.
  23. 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.
  24. 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.
  25. Zain J, Huang YQ, Feng X, Nierodzik ML, Li JJ, Karpatkin S: Concentration-dependent dual effect of thrombin on impaired growth/apoptosis or mitogenesis in tumor cells. Blood 2000;95:3133–3138.
  26. Hiraguri M, Miike S, Sano H, Kurasawa K, Saito Y, Iwamoto I: Granulocyte-macrophage colony-stimulating factor and IL-5 activate mitogen-activated protein kinase through Jak2 kinase and phosphatidylinositol 3-kinase in human eosinophils. J Allergy Clin Immunol 1997;100:S45–S51.
  27. Miike S, Nakao A, Hiraguri M, Kurasawa K, Saito Y, Iwamoto I: Involvement of JAK2, but not PI 3-kinase/Akt and MAP kinase pathways, in anti-apoptotic signals of GM-CSF in human eosinophils. J Leukoc Biol 1999;65:700–706.
  28. Kauffman HF, Tomee JF, van de Riet MA, Timmerman AJB, Borger P: Protease-dependent activation of epithelial cells by fungal allergens leads to morphologic changes and cytokine production. J Allergy Clin Immunol 2000;105:1185–1193.
  29. King C, Brennan S, Thompson PJ, Stewart GA: Dust mite proteolytic allergens induce cytokine release from cultured airway epithelium. J Immunol 1998;161:3645–3651.
  30. Lourbakos A, Potempa J, Travis J, D’Andrea MR, Andrade-Gordon P, Santulli R, Mackie EJ, Pike RN: Arginine-specific protease from Porphyromonas gingivalis activates protease-activated receptors on human oral epithelial cells and induces interleukin-6 secretion. Infect Immun 2001;69:5121–5130.
  31. Vu TK, Hung DT, Wheaton VI, Coughlin SR: Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991;64:1057–1068.
  32. Bohm SK, Kong W, Bromme D, Smeekens SP, Anderson DC, Connolly A, Kahn ML, Nelken NA, Coughlin SR, Payan DG, Bunnett NW: Molecular cloning, expression and potential functions of the human proteinase-activated receptor-2. Biochem J 1996;314:1009–1016.
  33. Ishihara H, Connolly AJ, Zeng D, Kahn ML, Zheng YW, Timmons C, Tram T, Coughlin SR: Protease-activated receptor 3 is a second thrombin receptor in humans. Nature 1997;386:502–506.
  34. Xu WF, Anderson H, Whitmore TE, Presnell SR, Yee DP, Ching A, Gilbert T, Davie EW, Foster DC: Cloning and characterization of human protease-activated receptor 4. Proc Natl Acad Sci USA 1998;95:6642–6646.
  35. Kahn ML, Nakanishi-Matsui M, Shapiro MJ, Ishihara H, Coughlin SR: Protease-activated receptor 1 and 4 mediate activation of human platelets by thrombin. J Clin Invest 1999;103:879–887.
  36. Nystedt S, Ramarkrishnan V, Sundelin J: The proteinase-activated receptor 2 is induced by inflammatory mediators in human endothelial cells. J Biol Chem 1996;271:14910–14915.
  37. Howells GL, Macey MG, Chinni C, Hou L, Fox MT, Harriott P, Stone SR: Proteinase-activated receptor-2: Expression by human neutrophils. J Cell Sci 1997;110:881–887.
  38. Mari B, Guerin S, Far DF, Breitmayer JP, Belhacene N, Peyron JF, Rossi B, Auberger P: Thrombin and trypsin-induced Ca(2+) mobilization in human T cell lines through interaction with different protease-activated receptors. FASEB J 1996;10:309–316.
  39. Mackie EJ, Pagel CN, Smith R, de Niese MR, Song SJ, Pike RN: Protease-activated receptors: A means of converting extracellular proteolysis into intracellular signals. IUBMB 2002;53:277–281.

    External Resources

  40. Vliagoftis H, Befus AD, Hollenberg MD, Moqbel R: Airway epithelial cells release eosinophil survival-promoting factors (GM-CSF) after stimulation of proteinase-activated receptor 2. J Allergy Clin Immunol 2001;107:679–685.
  41. Lindner JR, Kahn ML, Coughlin SR, Sambrano GR, Schauble E, Bernstein D, Foy D, Hafezi-Moghadam A, Ley K: Delayed onset of inflammation in protease-activated receptor-2-deficient mice. J Immunol 2000;165:6504–6510.
  42. Miike S, McWilliam AS, Kita H: Trypsin induces activation and inflammatory mediator release from human eosinophils through protease-activated receptor-2. J Immunol 2001;167:6615–6622.
  43. 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.
  44. Barrett AJ, Kirschke H: Cathepsin B, cathepsin H, and cathepsin L. Methods Enzymol 1981;80:535–561.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50