The success of immunological control methods is dependent upon the use of potential key antigens as tick vaccine candidates. Previously, we cloned a gene encoding 27 kDa and 30 kDa proteins (P27/30) of Haemaphysalis longicornis, and identified the P27/30 is a troponin I-like protein. In this study, the recombinant P27/30 (rP27/30) expressed in Escherichia coli was used to immunize mice and the mice were challenge-infested with ticks at different developmental stages of the same species. The rP27/30 protein stimulated a specific protective anti-tick immune response in mice, evidenced by the statistically significant longer pre-feeding periods in adult ticks. Furthermore, significantly longer feeding periods were noted in both larval and adult ticks. On the other hand, only larval ticks exhibited low attachment rates (31.1%). Immunization of mice with rP27/30 protein confers protection against hard tick Haemaphysalis longicornis infestation. These results demonstrated that the rP27/30 protein might be a useful vaccine candidate antigen for biological control of ticks.
Animals ; Feeding Behavior ; Female ; Ixodidae/*immunology ; Mice ; Mice, Inbred BALB C ; Microfilament Proteins/*immunology ; Recombinant Proteins/immunology ; Tick Infestations/*prevention&control ; Vaccines, Synthetic/immunology

The present study was conducted to identify new target immunogenic molecules from the larval stage of the cattle tick, Boophilus annulatus (Acari: Ixodidae). Two specific larval glycoproteins (GLPs) were isolated by two-step affinity chromatography. The larval immunogens were first purified with CNBr-Sepharose coupled to rabbit anti-larval immunoglobulins, and the glycoproteins were then purified with Con-A Sepharose. These glycoproteins have molecular weights of approximately 32 and 15 kDa with isoelectric points between 6.8 and 7.2. Antibodies against the two GLPs, labeled I and II, were detected in the anti-whole tick, -whole larval, and -gut antigens through immunoblot analysis. These results suggest that these GLPs are good immunogens and can be useful in the vaccination of cattle against tick infestation.
Amino Acid Sequence ; Animals ; Cattle ; Cattle Diseases/immunology/*parasitology/prevention & control ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Glycoproteins/immunology/*isolation & purification ; Immunoblotting ; Isoelectric Focusing ; Ixodidae/chemistry/*immunology ; Male ; Molecular Weight ; Rabbits ; Sequence Analysis, Protein ; Tick Infestations/immunology/parasitology/prevention & control/*veterinary

We investigated the induction of resistance to Haemaphysalis longicornis infestation in rabbits that had been immunized with recombinant H. longicornis P27/30 protein. The success of immunological control methods is dependent upon the use of potential key antigens as tick vaccine candidates. Previously, we cloned a gene encoding 27 kDa and 30 kDa proteins (P27/30) of H. longicornis, and identified P27/30 as a troponin I-like protein. In this study, rabbits that were immunized with recombinant P27/30 expressed in Escherichia coli showed the statistically significant longer feeding duration for larval and adult ticks (P< 0.05), low engorgement rates in larval ticks (64.4%), and an apparent reduction in egg weights, which suggest that H. longicornis P27/30 protein is a potential candidate antigen for a tick vaccine. These results demonstrated that the recombinant P27/30 protein might be a useful vaccine candidate antigen for biological control of H. longicornis.
Animals ; Antibodies/blood ; Escherichia coli/genetics ; Female ; Gene Expression ; Insect Proteins/immunology ; Ixodidae/*immunology ; Microfilament Proteins/*immunology ; Rabbits ; Recombinant Proteins/*immunology ; Research Support, Non-U.S. Gov't ; Tick Infestations/*immunology/prevention & control

In order to identify antigens that can help prevent camel tick infestations, three major glycoproteins (GLPs) about 97, 66 and 40 kDa in size were purified from adult and larval Egyptian ticks, Hyalomma (H.) dromedarii, using a single-step purification method with Con-A sepharose. The purified GLPs were evaluated as vaccines against camel tick infestation in rabbits. The rabbits received three intramuscular inoculations of GLPs (20 microg/animal) on days 0, 14, and 28. In the immunoblot analysis, Sera from the immunized rabbits recognized the native GLPs and other proteins from larval and adult H. dromedarii ticks along with those from other tick species such as Rhipicephalus sanguineus but not Ornithodoros moubata. The effects of immunity induced by these GLPs were determined by exposing rabbits to adult H. dromedarii ticks. These results demonstrated that GLP immunization led to a slightly decreased reproductive index and significantly reduced rates of egg hatchability. These results demonstrated that immunization with the purified GLPs can provide protection against infestation by H. dromedarii and some other tick species. Further studies are needed to confirm the effectiveness of immunization with GLPs against other tick species.
Animals ; Antigens/immunology/isolation & purification ; Argasidae/immunology ; Chromatography, Affinity/veterinary ; Electrophoresis, Polyacrylamide Gel/veterinary ; Female ; Glycoproteins/*immunology/isolation & purification ; Immunoblotting/veterinary ; Injections, Intramuscular/veterinary ; Ixodidae/growth & development/*immunology ; Life Cycle Stages ; Male ; Rabbits/*immunology/parasitology ; Reproduction ; Species Specificity ; Tick Infestations/immunology/prevention & control/*veterinary