用重组新型组织蛋白酶 B 对小鼠进行免疫接种可抑制旋毛虫发育，降低产卵量和虫荷。
Vaccination of mice with a recombinant novel cathepsin B inhibits Trichinella spiralis development, reduces the fecundity and worm burden.
机构信息
Department of Parasitology, Medical College, Zhengzhou University, Zhengzhou, 450052, China.
出版信息
Parasit Vectors. 2019 Dec 11;12(1):581. doi: 10.1186/s13071-019-3833-9.
BACKGROUND
Trichinella spiralis is a major zoonotic tissue-dwelling nematode, which is a public health concern and a serious hazard to animal food safety. It is necessary to exploit an anti-Trichinella vaccine to interrupt the transmission of Trichinella infection among animals and from animals to humans. The purpose of the present study was to characterize the novel T. spiralis cathepsin B (TsCB) and to evaluate the immune protection elicited by immunization with recombinant TsCB (rTsCB).
METHODS
The complete cDNA sequences of the TsCB gene were cloned, expressed and purified. The antigenicity of rTsCB was investigated by western blot analysis and ELISA. Transcription and expression of TsCB at various T. spiralis life-cycle stages were analyzed by RT-PCR and indirect immunofluorescent assay (IIFA). The mice were subcutaneously immunized with rTsCB, and serum level of TsCB-specific IgG (IgG1 and IgG2a) and IgE antibodies were assayed by ELISA. Immune protection elicited by vaccination with rTsCB was investigated.
RESULTS
The TsCB was transcribed and expressed in four T. spiralis life-cycle stages (adult worm, AW; newborn larvae, NBL; muscle larvae, ML; and intestinal infective L1 larvae), it was primarily located in the cuticle and stichosome of the parasitic nematode. Vaccination of mice with rTsCB produced a prominent antibody response (high level of specific IgG and IgE) and immune protection, as demonstrated by a 52.81% AW burden reduction of intestines at six days post-infection (dpi) and a 50.90% ML burden reduction of muscles at 35 dpi after oral larva challenge. The TsCB-specific antibody response elicited by immunization with rTsCB also impeded intestinal worm growth and decreased the female fecundity.
CONCLUSIONS
TsCB might be considered as a novel potential molecular target to develop vaccines against T. spiralis infection.
背景
旋毛虫是一种重要的人畜共患组织寄生线虫，对动物食品安全和公共卫生构成严重威胁。开发抗旋毛虫疫苗以阻断动物之间以及动物向人类传播旋毛虫感染是十分必要的。本研究旨在对旋毛虫组织蛋白酶 B（TsCB）进行特征描述，并评估重组 TsCB（rTsCB）免疫接种引发的免疫保护作用。
方法
克隆、表达和纯化了 TsCB 的完整 cDNA 序列。通过 Western blot 分析和 ELISA 研究了 rTsCB 的抗原性。通过 RT-PCR 和间接免疫荧光检测（IFA）分析了 TsCB 在旋毛虫不同生活史阶段的转录和表达。通过皮下免疫 rTsCB 对小鼠进行免疫接种，并通过 ELISA 检测 TsCB 特异性 IgG（IgG1 和 IgG2a）和 IgE 抗体的血清水平。研究了 rTsCB 疫苗接种引发的免疫保护作用。
结果
TsCB 在旋毛虫的四个生活史阶段（成虫、新生幼虫、肌肉幼虫和肠感染性 L1 幼虫）中均有转录和表达，它主要位于寄生线虫的角质层和纹皮肌层。rTsCB 疫苗接种可引起显著的抗体反应（高水平的特异性 IgG 和 IgE）和免疫保护，在口服幼虫攻毒后 6 天，肠道成虫负荷减少 52.81%，35 天肌肉幼虫负荷减少 50.90%。rTsCB 免疫接种引发的 TsCB 特异性抗体反应还抑制了肠道蠕虫的生长并降低了雌性的繁殖力。
结论
TsCB 可能被视为开发抗旋毛虫感染疫苗的一种新的潜在分子靶标。
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