Trials to Induce Protective Immunity in Mice and Sheep by Application of Protoscolex and Hydatid Fluid Antigen or Whole Body Antigen of Echinococcus granulosus

Abstract

In the current study, soluble proteins prepared from 200 mature Echinococcus granulosus and protoscolices of sheep hydatid cysts were applied to immunize sheep and mice respectively. The samples were mechanically homogenized in a blender, sonicated and the final yield was maintained at -20 degrees C until analysis. Hydatid fluid was isolated from liver or lung of sheep under sterile conditions. In the first experiment, 15 mice were randomly allocated to three groups of five mice each. Each mouse in groups 1 and 2 was immunized with 100 microg of hydatid fluid and protoscolex proteins in 100 microl of phosphate-buffered saline (PBS) and emulsified with an equal volume of Freund's complete adjuvant (FCA) respectively. The mice of group 3 were immunized with adjuvant in PBS. The mice were boosted 4 weeks after the first vaccination with the same preparation except that FCA was replaced by Freund's incomplete adjuvant (FIA). In the second experiment, eight male or female lambs 4-6 months of age, were allocated to two groups of four lambs each. Each lamb in the test group was vaccinated subcutaneously in the neck with a 2-ml dose of vaccine (1 mg of whole body protein of E. granulosus dissolved in 1 ml of PBS plus 1 ml of FCA). Control lambs were vaccinated with adjuvant in PBS. Lambs were boosted the same way as in the first experiment. Three weeks after the second vaccination, each mouse and lamb received a challenge infection with 2000 protoscolices intraperitoneally and each lamb additionally received 10 gravid E. granulosus. All mice and sheep were killed after 7 months and examined for hydatid cysts. In these studies, protective immunity was induced in mice with protoscolex protein and with hydatid fluid, and in sheep with whole-body homogenate of E. granulosus and the levels of protection afforded were found to be 72.1, 82.6 and 90.9% respectively.

Methods for samples preparation in proteomic research

“…Blenders can be used to dry or liquified samples, and in case of non-satisfactory results, sample usually needs to be processed by another apparatus, e.g. ultrasonic homogenizer, to receive optimal homogeneity [82]. This observation confirms the investigations of the homogenization method of the skeletal muscles to receive high enzymatic activity in the final solution.…”

Sample preparation and fractionation for proteome analysis and cancer biomarker discovery by mass spectrometry

“…In case of nonsatisfactory results, another type of homogenizers (i.e., ultrasonic) could be employed to achieve an optimal homogeneity [15].…”

Protective Properties of Echinococcus Multilocularis Protoscolex Antigens in Combination With the Immunomodulator Roncoleukin in Secondary Alveolar Echinoccocosis

“…Наши данные фактически не расходятся с выводами других исследователей, использовавших при гельминтозах в качестве средств потенциирования защитных механизмов организма животных различные иммуномодуляторы и адъювантные средства, повышающие иммуногенность антигенов гельминтов [10, 15,16,19,22].…”