Fawzi Elshaima M, González-Sánchez María Elena, Corral María Jesús, Alunda José Ma, Cuquerella Montserrat
Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Vet Parasitol. 2015 Jun 30;211(1-2):54-9. doi: 10.1016/j.vetpar.2015.04.029. Epub 2015 May 8.
Gene encoding a somatic protein of Haemonchus contortus (Hc23) known to confer significant protection against experimental haemonchosis has been cloned and expressed in a prokaryotic system. A cDNA library of H. contortus using the vector λ ZAP II was obtained. Full-length gene was amplified, cloned and expressed in Escherichia coli BL21. The recombinant protein was purified in Ni-NTA column. Recombinant protein (rHc23) had 203 aminoacids and a molecular mass of 24.15 kDa. Recombinant protein (100 μg/dose) with aluminum hydroxide as adjuvant was administered to 5-6 months age female Assaf lambs on days -42, -28 and -14. On day 0 animals were infected with 15,000 L3 of H. contortus. Vaccination with rHc23 elicited a significant protection against challenge, with >80% reductions in both fecal egg counts and average abomasal parasite burdens at the end of the experiment (45 days post challenge) besides lack of variations in packed cell volume. Results support the feasibility of vaccination against lamb haemonchosis with a recombinant product from an exposed antigen.
编码捻转血矛线虫（Hc23）一种体细胞蛋白的基因，已知该蛋白能对实验性血矛线虫病提供显著保护，已在原核系统中克隆并表达。利用λZAP II载体构建了捻转血矛线虫的cDNA文库。全长基因经扩增、克隆后在大肠杆菌BL21中表达。重组蛋白在镍-氮三乙酸（Ni-NTA）柱上纯化。重组蛋白（rHc23）含有203个氨基酸，分子量为24.15 kDa。以氢氧化铝为佐剂的重组蛋白（100μg/剂量）在第-42、-28和-14天给5-6月龄的阿萨夫母羊接种。在第0天，动物感染15000条捻转血矛线虫的L3期幼虫。用rHc23疫苗接种可引发显著的抗攻击保护作用，在实验结束时（攻击后45天），粪便虫卵计数和皱胃寄生虫平均负荷均降低了80%以上，此外红细胞压积无变化。结果支持了用暴露抗原的重组产物对羔羊血矛线虫病进行疫苗接种的可行性。
