• 1

    Yokogawa M, 1982. Paragonimiasis. Steele JH, eds. CRC Handbook Series in Zoonoses. Section C. Parasitic Zoonoses. Volume 3. Boca Raton, FL: CRC Press, 123–142.

  • 2

    Shim YS, Cho SY, Han YC, 1991. Pulmonary paragonimiasis: a Korean perspective. Semin Respir Med 12 :35–45.

  • 3

    Toscano C, Yu SH, Nunn P, Mott KE, 1995. Paragonimiasis and tuberculosis, diagnostic confusion: a review of the literature. Trop Dis Bull 92 :R1–R26.

    • Search Google Scholar
    • Export Citation
  • 4

    McKerrow JH, Sun E, Rosenthal PJ, Bouvier J, 1993. The proteases and pathogenicity of parasitic protozoa. Annu Rev Biochem 47 :821–853.

  • 5

    North MJ, Cotter DA, Franek KJ, 1990. Cysteine proteinase changes during macrocyst formation in Dictyostelium mucoroides.FEMS Microbiol Lett 60 :153–157.

    • Search Google Scholar
    • Export Citation
  • 6

    Chappell CL, Dresden MH, 1987. Purification of cysteine proteinases from adult Schistosoma mansoni.Arch Biochem Biophys 256 :560–568.

  • 7

    Rege AA, Herrera PR, Lopez M, Dresden MH, 1989. Isolation and characterization of a cysteine proteinase from Fasciola hepatica adult worms. Mol Biochem Parasitol 35 :89–96.

    • Search Google Scholar
    • Export Citation
  • 8

    Rosental PJ, McKerrow JH, Aikawa M, Nagasawa H, Leech JH, 1988. A malarial cysteine proteinase is necessary for hemoglobin degradation by Plasmodium falciparum.J Clin Invest 82 :1560–1566.

    • Search Google Scholar
    • Export Citation
  • 9

    Goldberg DE, Slater AF, Cerami A, Henderson GB, 1990. Hemoglobin degradation in the malaria parasite Plasmodium falciparum: an ordered process in a unique organelle. Proc Natl Acad Sci USA 87 :2931–2935.

    • Search Google Scholar
    • Export Citation
  • 10

    Brady CP, Dowd AJ, Brindley PJ, Ruan T, Day SR, Dalton JP, 1999. Recombinant expression and localization of Schistosoma mansoni cathepsin L1 support its role in the degradation of host hemoglobin. Infect Immun 67 :368–374.

    • Search Google Scholar
    • Export Citation
  • 11

    Chung YB, Kong Y, Joo IJ, Cho SY, Kang SY, 1995. Excystment of Paragonimus westermani metacercariae by endogenous cysteine protease. J Parasitol 81 :137–142.

    • Search Google Scholar
    • Export Citation
  • 12

    Chung YB, Yang HJ, Kang SY, Kong Y, Cho SY, 1997. Activities of different cysteine proteases of Paragonimus westermani in cleaving human IgG. Korean J Parasitol 35 :139–142.

    • Search Google Scholar
    • Export Citation
  • 13

    Hamajima F, Yamamoto M, Tsura S, 1994. Immunosuppression by neutral thiol protease from parasitic helminth larvae in mice. Parasite Immunol 16 :261–273.

    • Search Google Scholar
    • Export Citation
  • 14

    Kang SY, Cho MS, Chung YB, Kong Y, Cho SY, 1995. A cysteine protease of Paragonimus westermani eggs. Korean J Parasitol 33 :323–330.

  • 15

    Yamakami K, Hamajima F, Akao S, Tadakuma T, 1995. Purification and characterization of acid cysteine protease from metacercariae of the mammalian trematode parasite Paragonimus westermani.Eur J Biochem 233 :490–497.

    • Search Google Scholar
    • Export Citation
  • 16

    Dowd AJ, Smith AM, McGonigle S, Dalton JP, 1994. Purification and characterisation of a second cathepsin L proteinase secreted by the parasitic trematode Fasciola hepatica.Eur J Biochem 223 :91–98.

    • Search Google Scholar
    • Export Citation
  • 17

    Kong Y, Chung YB, Cho SY, Kang SY, 1994. Cleavage of immunoglobulin G by excretory-secretory cathepsin S-like protease of Spirometra mansoni plerocercoid. Parasitology 109 :611–621.

    • Search Google Scholar
    • Export Citation
  • 18

    Park H, Hong KM, Sakanari JA, Choi JH, Park SK, Kim KY, Hwang HA, Paik MK, Yun KJ, Shin CH, Lee JB, Ryu JS, Min DY, 2001. Paragonimus westermani: cloning of a cathepsin F-like cysteine proteinase from the adult worm. Exp Parasitol 98 :223–227.

    • Search Google Scholar
    • Export Citation
  • 19

    Cho SY, Hong ST, Rho YH, Choi S, Han YC, 1981. Application of micro-ELISA in serodiagnosis of human paragonimiasis. Korean J Parasitol 19 :151–156.

    • Search Google Scholar
    • Export Citation
  • 20

    Knobloch J, Lederer I, 1983. Immunodiagnosis of human paragonimiasis by an enzyme immunoassay. Trop Med Parasitol 34 :21–23.

  • 21

    Yokogawa M, Kojima S, Kobayashi M, Hata H, Ito J, Tsuji M, Miranda H, Ibanez N, Fernandez E, Guerra A, 1983. Peruvian paragonimiasis: diagnostic value of the enzyme-linked immunosorbent assay (ELISA). Jpn J Parasitol 32 :317–322.

    • Search Google Scholar
    • Export Citation
  • 22

    Imai J, 1987. Evaluation of ELISA for the diagnosis of paragon-imiasis westermani. Trans R Soc Trop Med Hyg 81 :3–6.

  • 23

    Gilmor SA, Cohen FE, 1993. New strategies for pharmaceutical design. Receptor 3 :155–163.

  • 24

    Kuntz ID, 1992. Structure-based strategies for drug design and discovery. Science 257 :1078–1082.

  • 25

    Sun E, Cohen FE, 1993. Computer-assisted drug discovery — a review. Gene 137 :127–132.

  • 26

    Gubler U, Hoffman BJ, 1983. A simple and very efficient method for generating cDNA libraries. Gene 25 :263–269.

  • 27

    Dalton JP, Clough KA, Jones MK, Brindley PJ, 1996. Characterization of the cathepsin-like cysteine proteinases of Schistosoma mansoni.Infect Immun 64 :1328–1334.

    • Search Google Scholar
    • Export Citation
  • 28

    Wex T, Wex H, Bromme D, 1999. The human cathepsin F gene–a fusion product between an ancestral cathepsin and cystatin gene. Biol Chem 380 :1439–1442.

    • Search Google Scholar
    • Export Citation
  • 29

    Maleewong W, Pariyanonda S, Wongkham C, Intapan P, Daenseegaew W, Morakote N, 1990. Comparision of adult somatic and excretory-secretory antigens in enzyme-linked immunosorbent assay for serodiagnosis of human infection with Paragonimus heterotremus.Trans R Soc Trop Med Hyg 84 :840–841.

    • Search Google Scholar
    • Export Citation
  • 30

    Ikeda T, Oikawa Y, Owhashi M, Nawa Y, 1992. Parasite-specific IgE and IgG levels in the serum and pleural effusion of paragonimiasis westermani patients. Am J Trop Med Hyg 47 :104–107.

    • Search Google Scholar
    • Export Citation
  • 31

    Ikeda T, Oikawa Y, Nishimura T, 1996. Enzyme-linked immunosorbent assay using cysteine proteinase antigens for immunodiagnosis of human paragonimiasis. Am J Trop Med Hyg 55 :435–437.

    • Search Google Scholar
    • Export Citation
  • 32

    Yamasaki H, Aoki T, Oya H, 1989. A cysteine proteinase from the liver fluke Fasciola spp.; purification, characterization, localization and application to immunodiagnosis. Jpn J Parasitol 38 :373–384.

    • Search Google Scholar
    • Export Citation
  • 33

    Chappell CL, Dresden MH, Gryseels B, Deelder AM, 1990. Antibody response to Schistosoma mansoni adult worm cysteine proteinases in infected individuals. Am J Trop Med Hyg 42 :335–341.

    • Search Google Scholar
    • Export Citation
  • 34

    Song CY, Kim TS, 1994. Characterization of cysteine proteinase from adult worms of Paragonimus westermani.Korean J Parasitol 32 :231–241.

    • Search Google Scholar
    • Export Citation
  • 35

    Maizel RM, Bundy DA, Selkirk ME, Smith DF, Anderson RM, 1993. Immunological modulation and evasion by helminth parasites in human population. Nature 365 :797–805.

    • Search Google Scholar
    • Export Citation
 
 
 

 

 
 
 

 

 

 

 

 

 

CLONING AND CHARACTERIZATION OF A NEW CYSTEINE PROTEINASE SECRETED BY PARAGONIMUS WESTERMANI ADULT WORMS

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  • 1 Departments of Internal Medicine, Pathology, and Parasitology, College of Medicine, Department of Pathology, College of Oriental Medicine, and Division of Biological Science, College of Natural Sciences, Wonkwang University, Iksan, Korea; College of Veterinary Medicine, Chonbuk National University, Jeonju, Korea; Department of Parasitology, College of Medicine, Chosun University, Kwangju, Korea

The cysteine proteinases of Paragonimus westermani are known to play important roles in invasion and pathogenesis to hosts and in immune modulation and nutrient uptake. In this study, we have cloned a new cysteine proteinase of P. westermani, PwCP2, from adult worms and tested its diagnostic usefulness. The PwCP2 gene had an open reading frame of 816 base pairs and a conserved catalytic triad of cysteine, histidine, and asparagine residues. The mature form of recombinant PwCP2 (rPwCP2) lacking a proregion was overexpressed in Escherichia coli and used to produce antiserum. Western blot and immunohistochemical analyses using this antiserum showed that PwCP2 was expressed as a mature form, 24-kD product in a crude extract and in the excretory-secretory product of P. westermani, and was localized mainly in the intestinal epithelium of the adult worm. Western blot analysis using the rPwCP2 showed not only high sensitivity (90%) and specificity (100%) to sera from patients with paragonimiasis westermani, but also no cross-reactivity with sera from patients with clonorchiasis, sparganosis, or cysticercosis. Furthermore, an enzyme-linked immunosorbent assay using rPwCP2 exhibited a sensitivity of 93% and a specificity of 93% with sera of rats infected with P. westermani metacercariae. These results suggest that the excretory-secretory PwCP2 can be used for the diagnosis of paragonimiasis.

Author Notes

Reprint requests: Hyun Park, Department of Parasitology, College of Medicine, Wonkwang University, Iksan 570-749, Korea, Telephone: 82-63-850-6769, Fax: 82-63-857-0342 E-mail: hyunpk@wonkwang.ac.kr.
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