Heilongjiang Key Laboratory for Zoonosis, College of Veterinary Medicine, Northeast Agricultural University, 600 Changjiang Street, Harbin, 150030, China.
Vet Res. 2021 Mar 25;52(1):51. doi: 10.1186/s13567-021-00921-1.
Trichinella spiralis is an important foodborne parasitic nematode distributed worldwide that infects humans and animals. Glutaminase (GLS) is an important gene in the glutamine-dependent acid resistance (AR) system; however, its role in T. spiralis muscle larvae (ML) remains unclear. The present study aimed to characterize T. spiralis GLS (TsGLS) and assess its function in T. spiralis ML AR both in vitro and in vivo using RNA interference. The results indicated that native TsGLS (72 kDa) was recognized by anti-rTsGLS serum at the muscle larvae stage; moreover, an immunofluorescence assay confirmed that TsGLS was located in the epidermis of ML. After silencing the TsGLS gene, the relative expression of TsGLS mRNA and the survival rate of T. spiralis ML were reduced by 60.11% and 16.55%, respectively, compared to those in the PBS and control groups. In vivo AR assays revealed that the worm numbers at 7 and 35 days post-infection (dpi) decreased by 61.64% and 66.71%, respectively, compared to those in the PBS group. The relative expression of TsGLS mRNA in F generation T. spiralis ML was reduced by 42.52%, compared to that in the PBS group. To the best of our knowledge, this is the first study to report the presence of the glutamine-dependent AR system in T. spiralis. Our results indicate that TsGLS plays a crucial role in the T. spiralis AR system; thus, it could be used as a potential candidate target molecule for producing vaccines against T. spiralis infection.
旋毛虫是一种分布广泛的重要食源性寄生线虫，可感染人类和动物。谷氨酰胺酶（GLS）是谷氨酰胺依赖的酸抗性（AR）系统中的一个重要基因；然而，其在旋毛虫肌肉幼虫（ML）中的作用尚不清楚。本研究旨在通过 RNA 干扰在体外和体内对旋毛虫 GLS（TsGLS）进行特征描述，并评估其在旋毛虫 ML AR 中的功能。结果表明，天然 TsGLS（72 kDa）在肌肉幼虫阶段被抗 rTsGLS 血清识别；此外，免疫荧光测定证实 TsGLS 位于 ML 的表皮中。沉默 TsGLS 基因后，与 PBS 组和对照组相比，TsGLS mRNA 的相对表达量和旋毛虫 ML 的存活率分别降低了 60.11%和 16.55%。体内 AR 测定显示，感染后 7 和 35 天的虫体数量分别减少了 61.64%和 66.71%，与 PBS 组相比。F 代旋毛虫 ML 中 TsGLS mRNA 的相对表达量比 PBS 组降低了 42.52%。据我们所知，这是首次报道旋毛虫存在谷氨酰胺依赖的 AR 系统。我们的结果表明 TsGLS 在旋毛虫 AR 系统中发挥着重要作用；因此，它可能成为旋毛虫感染疫苗的潜在候选靶标分子。
