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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

Abdul O. Mongi  and
C. A. Aganyo
Abdul O. Mongi
Affiliation:
The International Centre of Insect Physiology and Ecology (ICIPE), P. O. Box 30772, Nairobi, Kenya
C. A. Aganyo
Affiliation:
The International Centre of Insect Physiology and Ecology (ICIPE), P. O. Box 30772, Nairobi, Kenya
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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

Rhipicephalus appendiculatusmidgut antigensresistantrabbits immunoglobulinsimmunoblottingimmunoaffinityimmunizationimmunodiffusionion exchangenative proteinprotein subunitsimmunization skin tests
Type
Research Articles
Information
International Journal of Tropical Insect Science , Volume 11 , Issue 6 , December 1990 , pp. 907 - 916
Copyright
Copyright © ICIPE 1990

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