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Inmunogenicidad de la proteína recombinante ASP1R de ancylostoma caninum en un modelo murino.

Inmunogenicidad de la proteína recombinante ASP1R de ancylostoma caninum en un modelo murino.



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Giraldo G, M., & Castaño, J. (2009). Inmunogenicidad de la proteína recombinante ASP1R de ancylostoma caninum en un modelo murino. Revista MVZ Córdoba, 14(2). https://doi.org/10.21897/rmvz.349

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Maria Giraldo G
Jhon Castaño

Objetivo. Construir un plásmido recombinante que exprese la proteína ASP1r de Ancylostoma caninum y evaluar su capacidad inmunogénica en un modelo murino. Materiales y métodos. Se realizó extracción de ARN de parásitos adultos de Ancylostoma caninum, se amplificó por RT –PCR el gen de la proteína ASP1. Este gen fue insertado en el vector pcDNA3. El inserto fue digerido con Bamh1 y EcoR1 y clonado direccionalmente. Posteriormente, se llevó a cabo transformación y selección de las células de E. coli DH5a competentes con el producto de la ligación. Se realizó un tamizaje por PCR confirmando la presencia del gen ASP1. El vector pcDNA3-ASP1 fue administrado vía intraglandular en la parotida e intramuscular en ratones Balb/c. En estos animales se les realizó determinación de anticuerpos en suero y saliva mediante las técnicas de ELISA e inmunohistoquímica. Resultados. Se determinó que el plásmido pcDNA3-ASP1 fue incorporado y expresado células E. coli DH5a. Este plásmido recombinante indujo la producción de anticuerpos Anti-ASP1 específicos en ratones Balb/c. Conclusiones. Se logró demostrar que la utilización de pcDNA3-ASP1 no produjo reacciones desfavorables en ratones Balb/c, además indujo respuesta humoral contra la proteína pcDNA3-ASP1 de excreción/secreción de Ancylostoma caninum en ratones.

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  1. Cordero Del Campillo, M et al. Parasitología veterinaria. Madrid, Espa-a; McGraw–Hill: 2000.
  2. Hawdon JM, Hotez PJ.Hookwoorm: developmental biology of the infectious process. Curr Opin Gen Surg 1996; 6:618- 23. http://dx.doi.org/10.1016/S0959-437X(96)80092-X
  3. Hawdow JM, Narasimhan S, Hotez PJ. Ancylostoma secreted protein 2: Cloning and characterization of a second member of a family of nematode secreted proteins from Ancylostoma caninum. Mol Biochem parasitol 1999; 99:149-65. http://dx.doi.org/10.1016/S0166-6851(99)00011-0
  4. Hawdon JM, Schad GA.Serum stimulated feeding in vitro by 3 stage infective larvae of canine hookworm Ancylostoma caninum. J Parasitol 1990;76:394-8. http://dx.doi.org/10.2307/3282673
  5. Hawdon JM, Jones BF, and Hotez PJ.. Cloning and characterization of a cDNA encoding the catalytic subunit of a c AMPdependient protein kinase from Ancylostoma caninum third-stage infective larvae. Mol Biochem Parasitol 1995;69 :127-30. http://dx.doi.org/10.1016/0166-6851(94)00203-Y
  6. Hawdom JM, Jones BF, Perre gaux MA, Hotez PJ. Ancylostoma caninum metaloprotease release coincides with activation of infective larvae in vitro. Exp Parasitol 1995;80: 2005-11.
  7. Zhan B, Hawdon J, Shan Q, Ren HN, Quiang HQ, Hu W, et al. Ancylostoma secreted protein 1 (ASP-1) homologues in human hook worms. Mol Biochem Parasitol 1999; 98:143-9. http://dx.doi.org/10.1016/S0166-6851(98)00157-1
  8. Hawdow JM, Jones Bf, Hoffman DR, Hotez PJ. Cloning and characterization of Ancylostoma secreted protein: a novel protein associated with the transition to parasitism by infective hookworm larvae. J Biol Chem 1996;271:6672-8.
  9. Zhan B, Hotez PJ, Wang Y, Hawdon J. A developmentally regulated metalloproteasa secreted by host-stimulated Ancylostoma caninum third- stage infective larvae is a member of the astacin family of proteases. Mol Biochem Parasitol 2002; 120: 291-6. http://dx.doi.org/10.1016/S0166-6851(01)00453-4
  10. Ding X, Shields J, Allen R, Hussey RSMolecular cloning and caracterization of a vemon allergen AG5- like cDNA from Meloidogyne incognita. Int J Parasitol 2000; 30: 77-81.
  11. Gao B, Allen R, Maier T, Davis EL, Braum TJ, Hussey RS. Molecular characterization and expression of two venom allergen-like protein genes in Heterodera glycines. Int Parasitol 2001; 31: 1617-25. http://dx.doi.org/10.1016/S0020-7519(01)00300-9
  12. Tawe W, Pearlman E, Unnasch TR, Lustigman SAngiogenic activity of Onchocerca volvulus recombinant proteins similar to vespid venom antigen 5. Mol Biochem Parasitol 2000; 109:91-9. http://dx.doi.org/10.1016/S0166-6851(00)00231-0
  13. Hotez PJ, Ghosh K, Hawdon JM, Narasimhan S, Jones B, Xiao SH, et alExperimental approaches to the development of a recombinant hookworm vaccine. Immunol Rev 1999; 171: 163-71.
  14. Hotez PJ , Zhan B , Bethony J M , Loukas A , Williamson A , Goud G N , et al. Progress in the development of a recombinant vaccine for human hookworm disease: the Human Hookworm Vaccine Initiative. Int J Parasitol 2003; 33:1245-58. http://dx.doi.org/10.1016/S0020-7519(03)00158-9
  15. Schalling H, Va Leewen M, Verstrepen B, Cornelissen A. Molecular characterization and expression of two putative protective excretory secretory proteins of Haemonchus contortus. Mol Biochem Parasitol 1997; 88:203-13. http://dx.doi.org/10.1016/S0166-6851(97)00093-5
  16. De Maere V, Vercauteren I, Saverwyns H, Claerebout E, Bern G, Vercruysse J. Identification of potential protective antigens of Ostertagia ostertagi with local antibody probes. Parasitology 2002;125: 383- 91. http://dx.doi.org/10.1017/S0031182002002196
  17. Ghosh K, Hawdon J, Hopez P. Vaccinacion with alum-precipitated recombinant Ancylostoma secreted protein 1 protects mice against challenge infeccion with infective hookworm (Ancylostoma caninum) larvae. J Infect Dis 1996;174: 1380-3. http://dx.doi.org/10.1093/infdis/174.6.1380
  18. Ghosh K, Hopez JP. Antibody- dependent reduccion in mouse hookworm burden after vaccination with Ancylostoma caninum secreted protein 1 (Ac- ASP-1). J Infect Dis 1999; 180: 1674-81. http://dx.doi.org/10.1086/315059
  19. Liu S, Gosh K; Zhan B, Shan Q, Thompson MG, Hawdon J, et al. Hookworm burden reductions in Balb/c mice vaccinated with Ancylostoma secreted protein 1 (ASP-1) from Ancylostoma duodenale, A. caninum and Necator americanus. Vaccine 2000;18: 1096-02.
  20. Sharp PJ, Wagland BM, inventors. Nematode vaccine.US Patent 5 734 035, 1998 March 31.
  21. Murray J, Gregory WF, Gomez- Escobar N, Atmadja AK, Maizels RM. Expression and immune recognition of Brugia malayi VAL- 1, a homologue of vespid venom allergens and Ancylostoma secreted proteins. Mol Biochem Parasitol 2001;118: 89-96. http://dx.doi.org/10.1016/S0166-6851(01)00374-7
  22. Lustigman S, James ER, Tawe W, Abraham D. Towards a recombinant antigen vaccine against Onchocerca volvulus. Trends Parasitol 2002; 18:135–41. http://dx.doi.org/10.1016/S1471-4922(01)02211-5
  23. Sharp PJ, Wagland BM (Biotech Australia and CSIRO). Nematode Vaccine. International Patent Application Number PCT/AU92/00041. International Publication Number WO 92/13890.
  24. Schalling H, Va Leewen M, Hendrikx W. Immune responses of sheep to excretory/ secretory products of adult Haemonchus contortus. Parasitol 1994; 108:351-7. http://dx.doi.org/10.1017/S0031182000076198
  25. Schallig H, Van Leeuwen M.Protective immunity to the blood-feeding nematode Haemonchus contortus induced by vaccination with parasite low molecular weight antigens. Parasitology 1997; 114:293–99. http://dx.doi.org/10.1017/S0031182096008517
  26. Kooyman F, Schallig H, Van Leewen M, Mackellar A, Huntley JF, Cornelissen A, et al. Protection in lambs vaccinated with Haemonchus contortus antigens is age related, and correlated, with IgE rather than IgG1 antibody. Parasite Immunol 2000; 22:13-20. http://dx.doi.org/10.1046/j.1365-3024.2000.00265.x
  27. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual 3a edition, Vol. 1, 2 and 3. USA: Cold Spring Harbor Laboratory Press; 2001.
  28. Kerr KB. Studies on acquired immunity to the dog hookworm Ancylostoma caninum. Am J Hyg 1936; 23:381-406.
  29. Miller TA. Immunity to hookworm. In: Souls by EJL. ed, Immune responses in parasitic infections: immunology, immunopathology and immunoprophylaxis, Vol I, Nematodes, Boca Raton, FL: CRC Press; 1981.
  30. Sen L, Ghosh K, Bin Z, Qiang S, Thompson M, Hawdon J , et al. Hookworm burden reductions in BALB/c mice vaccinated with recombinant Ancylostoma secreted proteins (ASPs) from Ancylostoma duodenale, Ancylostoma caninum and Necator americanus. Vaccine 2000; 18:1096-1102. http://dx.doi.org/10.1016/S0264-410X(99)00371-0
  31. Mendez S, Zhan B, Goud G, Ghosh K, Dobardzic A, Wu W, et al. Effect of combining the larval antigens Ancylostoma secreted protein 2 (ASP-2) and metalloprotease 1 (MTP-1) in protecting hamsters against hookworm infection and disease caused by Ancylostoma ceylanicum. Vaccine 2005;23: 3123–30. http://dx.doi.org/10.1016/j.vaccine.2004.12.022
  32. Zhan B, Liu S, Perally S, Xue J, Fujiwara R, Brophy P et al. Biochemical Characterization and Vaccine Potential of a Heme Binding Glutathione Transferase from the Adult Hookworm Ancylostoma caninum. Infect immun 2005; 75:6903–6911. http://dx.doi.org/10.1128/IAI.73.10.6903-6911.2005

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