1921
Volume 71, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

The cysteine proteinases of 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 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 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 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 metacercariae. These results suggest that the excretory-secretory PwCP2 can be used for the diagnosis of paragonimiasis.

Loading

Article metrics loading...

/content/journals/10.4269/ajtmh.2004.71.87
2004-07-01
2017-09-24
Loading full text...

Full text loading...

/deliver/fulltext/14761645/71/1/0700087.html?itemId=/content/journals/10.4269/ajtmh.2004.71.87&mimeType=html&fmt=ahah

References

  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.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  13. Hamajima F, Yamamoto M, Tsura S, 1994. Immunosuppression by neutral thiol protease from parasitic helminth larvae in mice. Parasite Immunol 16 : 261–273.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  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.
  34. Song CY, Kim TS, 1994. Characterization of cysteine proteinase from adult worms of Paragonimus westermani. Korean J Parasitol 32 : 231–241.
  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.
http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.2004.71.87
Loading
/content/journals/10.4269/ajtmh.2004.71.87
Loading

Data & Media loading...

  • Received : 03 Mar 2003
  • Accepted : 18 Feb 2004

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error