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Rhipicephalus appendiculatus: Immunochemical isolation, identification and characterization of tick gut antigens recognized by IgGs derived from rabbits repeatedly infested with ticks
Published online by Cambridge University Press: 19 September 2011
Abstract
Immune resistance to infestation by the brown ear tick, Rhipicephalus appendiculatus Neumann, was induced in rabbits by repeated tick feeding. Antibodies from the rabbits resistant to ticks were purified and used to isolate tick midgut antigens. Purified immunoglobulin G (IgGs) from rabbits immune to tick infestations demonstrated, by immunoblotting, nine protein subunits with molecular weights ranging from 12,000 to 92,000 daltons. Immuno-adsorption studies revealed an aggregated native protein with a molecular weight greater than 500,000 daltons as resolved by polyacrylamide gel electrophoresis (PAGE). Sodium dodecyl sulphate (SDS)-PAGE of this protein molecule revealed the presence of 14 protein subunits similar to some of those detected by the immunoblots. Their molecular weights ranged from 14,000 to 165,000 daltons. Sera from rabbits immunized with the 500,000 dalton molecule formed a weak precipitating arc in the immunodiffusion test, confirming its immunogenicity. The reproductive potential of ticks fed on rabbits immunized with the purified protein molecule was slightly affected compared to controls. The protein molecule also had the capacity to induce immediate and delayed type skin reactions when injected into rabbits resistant to adult R. appendiculatus, as opposed to control tick-naive rabbits.
Résumé
La resistance immunitaire à l'infestation des tiques Rhipicephalus appendiculatus Neumann, a été induite sur des lapins après alimination repetée. Les anticorps des lapins resistants aux tiques furent purifies et utilisés pour isoler les antigenes de l'intestin des tiques. L'immunoglobuline G (IgGs) purifié des lapins immunisés à l'infestion des tiques a demontré à l'immunoblotting 9 sous unités de protéines avec un poids moleculaire allant de 12,000 à 92,000 daltons. L'immunoabsorption a relèvé une proteine avec un poids moleculaire plus grand que 500,000 daltons grace à l'électrophorese du gel polyacrylamide (PAGE). Le SDS-PAGE de cette molecule a relèvé la presence de 14 sous unités de proteines. Ces 14 proteines furent detectées à l'immunoblots. Le poids moleculaire de ces proteines se situait entre 14,000 et 165,000 daltons. Les serums des lapins immunizés avec la proteines au poids moleculaire 500,000 daltons forma un arc de precipitation au test de l'immunodiffusion pour confirme l'immunogenecité de cattée proteine. Le potentiel reproductive des tiques nourris sur des lapin immunizés avec la molecule de proteine purifiée a été legèrement affectée par rapport aux lapins utilisés comme contrôle. La molecule de proteine aussi la capacité d'induire des reactions cutanées immediates ou retardées lorsqu'elle est injectée aux lapins resistants aux adultes du R. appendiculatus a l'opposé des contrôles qui sont des lapins qui ne pas exposés aux tiques.
Keywords
- Type
- Research Articles
- Information
- International Journal of Tropical Insect Science , Volume 11 , Issue 6 , December 1990 , pp. 907 - 916
- Copyright
- Copyright © ICIPE 1990
References
REFERENCES
Agbede, R. I. S. and Kemp, D. H. (1986) Immunization of cattle against Boophilus microplus using extracts derived from adult female ticks: Histopathology of ticks feeding on vaccinated cattle. Int. J. Parasitol. 16, 35–41.CrossRefGoogle ScholarPubMed
Allen, J. R. and Humphreys, S. J. (1979) Immunization of guinea pigs and cattle against ticks. Nature (London) 280, 491–493.CrossRefGoogle ScholarPubMed
Bailey, K. P. (1960) Notes on the rearing of Rhipicephalus appendiculatus and their infection with Theileria parva for experimental transmission. Bull. Epizoot. Dis. Afr. 8, 33–43.Google Scholar
Binnington, K. C. and Kemp, D. H. (1980) Role of tick salivary glands in feeding and disease transmission. Adv. Parasitol. 18, 315–339.CrossRefGoogle ScholarPubMed
Brown, S. J., Shapiro, S. Z. and Askenase, P. W. (1984) Characterization of tick antigens inducing host immune resistance. I. Immunization of guinea pigs with Amblyomma americanum-derived salivary gland extract and identification of an important salivary gland protein antigen with guinea pig anti-tick antibodies. J. Immunol. 133, 3319–3325.CrossRefGoogle ScholarPubMed
Irvin, A. D. and Broadesky, D. W. (1970) Rearing and maintaining Rhipicephalus appendiculatus in the laboratory. J. Inst. Anim. Technol. 21, 106–112.Google Scholar
Johnston, L. A. Y., Kemp, D. H. and Pearson, R. D. (1986) Immunization of cattle against Boophilus microplus using extracts derived from adult female ticks: Effects of induced immunity to tick populations. Int. J. Parasitol. 16, 27–34.CrossRefGoogle ScholarPubMed
Kemp, D. H., Stone, B. F. and Binnington, K. C. (1982) Tick attachment and feeding: Role of the mouthparts, feeding apparatus, salivary gland secretions and the host response. In Physiology of Ticks (Edited by Obenchain, F. D. and Galun, R.), pp. 119–168. Pergamon Press, Oxford.CrossRefGoogle Scholar
Laemli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.CrossRefGoogle Scholar
Maizel, J. V. (1971) Polyacrylamide gel electrophoresis of viral proteins. Meth. Virol. 5, 179–246.Google Scholar
Merril, C. R., Goldman, D., Sedman, S. A. and Elbert, M. H. (1981) Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 21, 1437–1441.CrossRefGoogle Scholar
Mongi, A. O., Shapiro, S. Z., Doyle, J. J. and Cunningham, M. P. (1986a) Immunization of rabbits with Rhipicephalus appendiculatus antigen-antibody complexes. Insect Sci. Applic. 7, 471–477.Google Scholar
Mongi, A. O., Shapiro, S. Z., Doyle, J. J. and Cunningham, M. P. (1986b) Characterization of antigens from extracts of fed ticks using sera from rabbits immunized with extracted tick antigen and by successive tick infestation. Insect Sci. Applic. 1, 479–487.Google Scholar
Ouchterlony, O. (1958) Diffusion-in-gel methods for immunological analysis. Progress in Allergy 5, 1–78.Google ScholarPubMed
Porath, J. (1974) General methods and coupling procedures. in Methods in Enzymology XXXIV (Edited by Jakoby, W. B. and Wilcheck, M.), pp. 13–29. Academic Press, New York.CrossRefGoogle Scholar
Purnell, R. E. and Joyner, L. P. (1968) The development of Theileria parva in the salivary glands of the tick Rhipicephalus appendiculatus. Parasitology 58, 725–730.CrossRefGoogle ScholarPubMed
Towbin, H., Stachlin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Science (Washington) 76, 4350–4354.CrossRefGoogle ScholarPubMed
Wharton, R. H. (1976) Tick-borne livestock diseases and their vectors. V. Acaricide resistance and alternative methods of tick control. World Anim. Rev. 20, 8–15.Google Scholar
Wikel, S. K. and Allen, J. R. (1976) Acquired resistance to ticks. I. Passive transfer of resistance. Immunology 30, 311–316.Google ScholarPubMed
Wikel, S. K. and Allen, J. R. (1982) Immunological basis of host resistance to ticks. In Physiology of Ticks (Edited by Obenchain, F. D. and Galun, R.), pp. 169–196. Pergamon Press, Oxford.CrossRefGoogle Scholar
Willadsen, P. and Riding, G. A. (1979) Characterization of a proteolytic enzyme inhibitor with allergenic activity. Biochem. J. 17, 41–47.CrossRefGoogle Scholar