Production of monoclonal antibody to 45 kDa somatic protein of Trichuris suis

돼지편층의 45kDa 항원단백질에 대한 단클론항체 생산

  • Accepted : 2004.09.15
  • Published : 2004.12.30

Abstract

Trichnuris suis does not excrete eggs during larval stage as well as in particular adult stage, It is impossible to diagnose by use of fecal examination method in those periods. Therefore, serological diagnostic method can be very useful for those stages. In order to produce monoclonal antibody, specific somatic and secretory-excretory (SE) antigens of T. suis were identified and analyzed by SDS-PAGE and Western blot. Monoclonal antibody-producing hybridoma cells were cloned, which were made of popliteal lymph node of BALB/c mice immunized with a 45 kDa somatic antigen of T. suis. Five clones (1B9, 2C4, n2C5, 2D7 and 2D8) showing strong responses to T. suis antigens were selected and the isotype identified. All monoclonal antibodies were IgG1 isotype and the light chains were k chain. Established monoclonal antibodies reacted specifically to somatic and SE antigens of T. suis and did not cross-reacted to antigens of ascaris suum, trichuris vulpis, or Trichinella spiralis. The sensitivity of somatic and SE antigens against these monoclonal antibodies were significant (p<0.01) associated with those of positive and negative sera.

Keywords

References

  1. 강영배, 위성환, 김상희, 장환, 최상호. 최근 기업화양돈장의 돼지 내부기생충 감염실태 변화양상 조사. 한국수의공중보건학회지. 1989, 13, 15-19
  2. 권창희. Psedorabies Virus 단클론항체를 이응힌 개량효소면역확산법 개발 및 gP50과 gP63 유전자 클로닝에 관한 연구. 서울대학교 대학원 박사 학위 논문. 서울, pp. 43, 1991
  3. 이재구. 최신 수의 기생충학. pp. 266-268, 대한교과서주식회사. 서울, 1987
  4. 장두환, 재욱, 강두원. 발효톱밥 돈사에 대한 돼지내부기생충 조사. 대한수의학회지. 1991, 31, 509-513
  5. 지차호, 박승준. 시험관내에서 돼지회충(Ascaris suun) 함자충란($L_2$)의 인공배앙. 대한수의학회지. 1998, 38, 107-117
  6. 지차호, 이철순, 박승준. 혈청학적 진단을 위한 돼지편충의 체항원, 배설/분비항원의 분리 및 비교. 대한수의학회지. 1999, 39, 159-168
  7. 채창수. Sulfamethazine에 대한 단클론항체의 생산과 이를 이용한 ELISA의 개발. 서울대학교 대학원 석사학위 논문. 서울. pp. 6-10, 1996
  8. 홍영선. 단클론항체 이용 돼지 회충 분비 항원 및 체항원 특이성 분석. 서울대학교 대학원 박사 학위 논문. 서울, pp. 43-44, 1999
  9. Beer, R. J. S. Studies on the biology of the life-cycle of Trichuris suis Schrank, 1788. Parasitology. 1973, 67, 253-262
  10. Bj$\"{o}$rkman, C(amilla)., Joakim, O., Holmdahl, M. and Uggla, A. An indirect enzyme-linked immunoassay (ELISA) for demonstration of antibodies to Neospira caninum in serum and milk of cattle. Vet. Parasitol. 1997, 68, 251-260
  11. Chan, S. W. and Ko, R. C. Specificity of affinitypurified Trichinella spiralis antigens. Vet. Parasitol. 1992, 41, 109-120
  12. Deplazes, P., Gottstein, B., Stingelin, Y. and Eckert, J. Detection of Taenia hydatigena Copro-Antigens by ELISA in Dogs. Vet. Parasitol. 1990, 36, 91-103
  13. Drake, L. J., Bianco, A. E., Bundy, D. A. P. and Ashall, F. Characterization of peptidases of adult Trichuris muris. Parasitol. 1994, 109, 623-630
  14. Else, K. J., Wakelin, D., Wassom, D. L. and Hauda, K. M. MHC-restricted antibody responses to Trichuris muris secretory-excretory (SE) antigen. Parasite Immunol. 1990, 12, 509-527
  15. Fetterer, R. H. and Hill, D. E. Localization of phenol oxidase in female Trichuris suis. J. Parasitol. 1994, 80, 952-959
  16. Fetterer, R. H. and Hill, D. E. The occurrence of phenol oxidase activity in female Trichuris suis. J. Parasitol. 1993, 79, 155-159
  17. Fields, H. A., Bruguera, P., de la Torre, N., Puig, J. and Anderson, L. J. Purity, antigenicity and immunogenicity of the hepatitis B surface antigen purified by five different methods. J. Virol. Methods. 1988, 22, 283-294
  18. Greenspon, L. W., White, J., Shields, R. S., Funer, A. and Gold, W. M. Purification of Ascaris suum antigen: Its allergenic activity in vitro and in vivo. J Allergy Clin. Immunol. 1986, 77, 443-51
  19. Hill, D. E. Protective Antigens from culture Fluids of the Adult Swine Whipworm, Trichuris suis. Research Investment Report (National Pork Producers Council). 1996
  20. Hill, D. E., Fetterer, R. H., Romanowski, R. D. and Urban, J. F. Jr. The effect of immunization of pigs with Ascaris suum cuticle components on the development of resistance to parenteral migration during a challenge infection. Vet. Immunol. Immunopathol. 1994, 42, 161-169
  21. Hill, D. E., Romanowski, R. D. and Urban, J. F. Jr. A Trichuris specific diagnostic antigen from culture fluids of Trichuris suis adult worms. Vet. Parasitol. 1997, 68, 91-102
  22. Lillywhite, J. E., Cooper, E. S., Needham, C. S., Venugopal, S., Bundy, D. A. P. and Bianco, A. E. Identification and characterization of excreted/secreted products of Trichuris trichiura. Parasite Immunol. 1995, 17, 47-54
  23. Homan, W. L., Derksen, A. C. and van Knapen, F. Identification of diagnostic antigens from Trichinella spiralis. Parasitol. Res. 1992, 78, 112-119
  24. Needham, C. S., Bundy, D. A. P., Lillywhite, J. E., Didier, J. M., Simmons, I. and Bianco, A. E. The relationship between Trichuris trichiura transmission intensity and the age-profiles of parasite-specific antibody isotypes in two endemic communities. Parasitology. 1992, 105, 273-283
  25. Prichard, R. Application of molecular biology in veterinary parasitology. Vet. Parasitol. 1997, 71, 155-175
  26. Pupo, E., Aguila, A., Santana, H., Nunez, JF., Castellanos-Serra, L. and Hardy, E. Mice immunization with gel electrophoresis micropurified bacterial lipopolysaccharides. Electrophoresis. 1999, 20, 458-461
  27. Roach, T. I. A., Else, K. J., Wakelin, D., Mclaren, D. J. and Grencis, R. K. Trichuris muris: antigen recognition and transfer of immunity in mice by IgAmonoclonal antibodies. Parasite Immunol. 1991, 13, 1-12
  28. Roach, T. I. A., Wakelin, D., Else, K. J. and Bundy, D. A. P. Antigenic cross-reactivity between the human whipworm, Trichuris trichiura, and the mouse trichuroids Trichuris muris and Trichinella spiralis. Parasite Immunol. 1988, 10, 279-291
  29. Salah, F., Demerdash, Z., Shaker, Z., El Bassiouny, A., El Attar, G., Iamail, S., Badir, N., Saad El Din, A. and Mansour, M. A monoclonal antibody against Schistosoma haematobium soluble egg antigen: efficacy for diagnosis and monitoring of cure of S. haematobium infection. Parasitol. Res. 2000, 86, 74-80
  30. Urban, J. F., Alizadeh, H. and Romanowski, R. D. Ascaris suum : Development of intestinal immunity to infective second stage larvae in swine. Exp. Parasitol. 1988, 66, 66-77
  31. 平認 亨, 池 逸夫. 豚の內部奇生蟲病と對策. 畜産の硏究. 1990, 44, 601-608