SD雄鼠3种不同新蛋白构型GnRH去势抗原免疫学及生物学效果对比研究

doi:10.11843/j.issn.0366-6964.2019.04.007

Abstract

Abstract:

The study was aimed to study the immunological and biological efficacy of 3-different types of GnRH-like constructs in male Sprague Dawley (SD) rats. The designing strategies for the new GnRH-like peptide constructs are based on:1) increasing the number of GnRH copy; 2) increasing the molecular weight of GnRH antigen. The new GnRH-like peptide constructs were designed based on a GnRH-tandem peptide, which was modified by relacing Gly at position 6 of the decapeptide with D-Lys (G6KT) to allow the conjugation of the GnRH peptide to ovalbumin (OVA). The first new GnRH-like construct was prepared by the dimerization of G6KT (G6KTD); the second and third were prepared by conjugating G6KT and G6KTD to OVA, respectively (G6KT-OVA and G6KTD-OVA). Sixty male SD rats at the age of 6 weeks, were randomly allocated to 5 groups (n=12):intact control (no treatment), surgically castrated or vaccinated against 100 μg peptide equivalent of G6KTD, G6KT-OVA and G6KTD-OVA in Specol adjuvant, respectively, at 6 weeks of age (with a booster 8 weeks later). Serum anti-GnRH antibody titers and reproductive hormone concentrations were assayed by RIA and ELISA, respectively. mRNA expressions of reproduction-related genes in pituitary-testicular axis were quantified using qPCR.The results shown that:1) active immunization against all of the 3 different new GnRH-like constructs could intrigue a good antibody responses in male SD rats; 2) Only with the first injection, active immunization against both G6KT-OVA and G6KTD-OVA intrigued a high antibody generation(P<0.05) and reduced serum concentrations of LH, FSH, testosterone and inhibin B(P<0.05), and further caused antrophy of testes with completely disrupted spermatogenesis and downregulated mRNA expressions of pituitary GnRHR, LHβ, FSHβ and testicular LHR, FSHR, INHα, INβA and INHβB (P<0.05) with the booster injection; 3) in contrast, the significant decrease of serum concentrations of LH, FSH, testosterone and inhibin B in rats actively immunized against G6KTD only occurred after the booster injection(P<0.05), but which still caused antrophy of testes with obviously disrupted spermatogenesis at decapitation (P<0.05) in 9 of 12 (75%) rats. In conclusion,based on those results, we concluded that both G6KT-OVA and G6KTD-OVA were highly efficacious GnRH antigen, which could be used as one-shot immunocastration vaccine in the furthure, whereas G6KTD itself was also effective, which might be used as GnRH antigen so as to avoid the drawbacks derived from the conjugation and purification processes of GnRH-carrier conjugates. Our findings promoted the further development and large scale commercial applications of GnRH vaccine.

Key words: GnRH, new protein comformation, immunocastration, rat

CLC Number:

S814.1

HAN Xingfa, LI Junli, CAO Xiaohan, CHU Mingxing, ZENG Xianyin. Immunological and Biological Efficacy of Three-different Types of New GnRH-like Constructs in Male SD Rats[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 738-749.

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Immunological and Biological Efficacy of Three-different Types of New GnRH-like Constructs in Male SD Rats

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