acid-phosphatase and Tick-Infestations

The full-length cDNA encoding acid phosphatase (HL-3) from Haemaphysalis longicornis was obtained by 5' rapid amplification of cDNA ends (RACE). The cDNA contained a 1137 bp open reading frame (ORF) coding for 356 amino acids with a predicted theoretical isoelectric point (pI) of 6.35 and molecular weight of 41.0 kDa. The recombinant protein was expressed in Escherichia coli. The enzyme could hydrolyze para-nitrophenyl phosphate (pNPP) substrate at an optimum pH of 5.0. Real-time RT-PCR analysis showed that the HL-3 transcripts were expressed in various stages of unfed ticks and were significantly induced by blood feeding. Furthermore, the expression of HL-3 in midguts was significantly higher than in other tested tissues of partially fed adult ticks. The transcripts of the HL-3 mRNA in lipopolysaccharide (LPS)-injected ticks were 1.75 times of the PBS-injected control; Theileria sergenti infected larvae expressed 3.86 more times than that of uninfected ones. Western blot analysis showed that rabbit antiserum against the recombinant rHL-3 could recognize a native protein of approximately 41.0 kDa in the lysates from different stages of ticks. Vaccination of rabbits with the rHL-3 conferred partial protective immunity against ticks, resulting in 28% mortality and 10.6% reduction in engorgement weight of adult ticks, respectively. These results suggested that the HL-3 was involved in tick innate immunity and could be used as a potential candidate antigen for the development of anti-tick vaccines.

Topics: Acid Phosphatase; Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; Gene Expression Regulation, Enzymologic; Ixodidae; Models, Molecular; Molecular Sequence Data; Phylogeny; Protein Conformation; Rabbits; Tick Infestations; Vaccines

Topics: Acid Phosphatase; Animals; Antibody Formation; Cattle; Guinea Pigs; Microsomes; Phosphoric Monoester Hydrolases; Tick Infestations; Ticks

Topics: Acid Phosphatase; Animal Nutritional Physiological Phenomena; Animals; Biopsy; Capillaries; Cattle; Cattle Diseases; Ethinyl Estradiol; Ethisterone; Hair; Hydroquinones; Melanocytes; Monophenol Monooxygenase; Peroxidases; Phagocytes; Pigmentation; Skin; Tick Infestations