Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology; Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, Hefei, 230032, Anhui Province, People's Republic of China.
The Clinical Laboratory of the Third People's Hospital of Heifei, Hefei, China.
Parasit Vectors. 2021 May 13;14(1):253. doi: 10.1186/s13071-021-04752-z.
The predominant genotype of Toxoplasma in China is the Chinese 1 (ToxoDB#9) lineage. TgCtwh3 and TgCtwh6 are two representative strains of Chinese 1, exhibiting high and low virulence to mice, respectively. Little is known regarding the virulence mechanism of this non-classical genotype. Our previous RNA sequencing data revealed differential mRNA levels of TgMIC1 in TgCtwh3 and TgCtwh6. We aim to further confirm the differential expression of TgMIC1 and its significance in this atypical genotype.
Quantitative real-time PCR was used to verify the RNA sequencing data; then, polyclonal antibodies against TgMIC1 were prepared and identified. Moreover, the invasion and proliferation of the parasite in HFF cells were observed after treatment with TgMIC1 polyclonal antibody or not.
The data showed that the protein level of TgMIC1 was significantly higher in high-virulence strain TgCtwh3 than in low-virulence strain TgCtwh6 and that the invasion and proliferation of TgCtwh3 were inhibited by TgMIC1 polyclonal antibody.
Differential expression of TgMIC1 in TgCtwh3 and TgCtwh6 may explain, at least partly, the virulence mechanism of this atypical genotype.
中国刚地弓形虫的优势基因型为中国 1 型（ToxoDB#9）谱系。TgCtwh3 和 TgCtwh6 是中国 1 型的两个代表性菌株，分别对小鼠具有高毒力和低毒力。关于这种非典型基因型的毒力机制知之甚少。我们之前的 RNA 测序数据显示 TgMIC1 在 TgCtwh3 和 TgCtwh6 中的 mRNA 水平存在差异。我们旨在进一步证实 TgMIC1 的差异表达及其在这种非典型基因型中的意义。
使用定量实时 PCR 验证 RNA 测序数据；然后，制备和鉴定针对 TgMIC1 的多克隆抗体。此外，在不使用 TgMIC1 多克隆抗体或使用 TgMIC1 多克隆抗体处理后，观察寄生虫在 HFF 细胞中的入侵和增殖情况。
数据表明，高毒力株 TgCtwh3 中的 TgMIC1 蛋白水平明显高于低毒力株 TgCtwh6，TgMIC1 多克隆抗体抑制了 TgCtwh3 的入侵和增殖。
TgCtwh3 和 TgCtwh6 中 TgMIC1 的差异表达至少部分解释了这种非典型基因型的毒力机制。
