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Study on Immunization Methods against Swine Trichinellosis and Biochemical Characteristics of Trichinella Antigens

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1. Biochemical characteristics of Trichinella antigens: For the first time, advanced techniques including sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IEF), two-dimensional electrophoresis and enzyme-linked immunoelectrotransfer blot (EITB) were applied to analyze 4 types of Trichinella antigens. The analysis showed that the soluble antigen of muscle larvae contains at least glycoproteins, lipoproteins, glycolipoproteins, simple proteins, polysaccharides and esterase isozymes. Two-dimensional electrophoresis revealed 122 polypeptide spots, most of which were acidic polypeptides with molecular weights below 67 KD.

1. Biochemical characteristics of Trichinella antigens: For the first time, advanced techniques including sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IEF), two-dimensional electrophoresis and enzyme-linked immunoelectrotransfer blot (EITB) were applied to analyze 4 types of Trichinella antigens. The analysis showed that the soluble antigen of muscle larvae contains at least glycoproteins, lipoproteins, glycolipoproteins, simple proteins, polysaccharides and esterase isozymes. Two-dimensional electrophoresis revealed 122 polypeptide spots, most of which were acidic polypeptides with molecular weights below 67 KD.

The excretory-secretory (ES) antigen of muscle larvae contains at least glycoproteins, lipoproteins, glycolipoproteins, simple proteins and polysaccharides. Two-dimensional electrophoresis revealed 61 polypeptide spots, most of which were acidic polypeptides with molecular weights below 67 KD. After SDS-PAGE, the soluble antigens of adult worms and newborn larvae showed 26 and 11 protein bands, respectively.

2. Study on immunization methods against swine trichinellosis: The research showed that 7 types of antigens had good immunoprotective effects on mice and pigs. Among them, the soluble antigen of adult worms and inactivated newborn larvae had the best immunogenicity, inducing 95.6% and 91.38% immunity (worm reduction rate) respectively in mice; inducing 98.21% and 95.3% immunity respectively in pigs; and in regional trials, the protection rates for pigs reached 75.26% and 75.99% respectively.

Studies on the immune mechanism showed that the immunity induced in pigs after vaccination with antigens not only reduced the number of worms but also weakened infectivity. Immune serum could kill muscle larvae in vitro. After immunization, serum antibodies increased, B lymphocytes in peripheral blood decreased, and the total number of T lymphocytes increased, among which suppressor T cells increased more, showing a temporary phenomenon of cellular immunosuppression. The duration of immunity was at least 6 months.

The research screened out an immunization method using white oil-Span adjuvant to emulsify the soluble antigen of 7-day-old adult worms, administered by intraperitoneal injection. A soluble antigen vaccine of adult worms and an inactivated newborn larvae vaccine were preliminarily developed. Regarding the research results on the biochemical characteristics of Trichinella antigens and immunization methods against swine trichinellosis, the partial biochemical characteristics of 4 types of antigens were elucidated; the immune characteristics of the antigens were clarified; effective immunogens and immunization methods against swine trichinellosis were screened out; and two vaccines with good protective effects on pigs were preliminarily developed. These results filled a domestic gap, reached the domestic leading level and international advanced level, and have major academic significance and huge potential application prospects.

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