Shen Chunxiang, Zhu Xinyi, Xu Xuejun, Chang Hao, Ni Yangyue, Li Chen, He Kaiyue, Chen Lin, Chen Lu, Hou Min, Ji Minjun, Xu Zhipeng
Department of Pathogen Biology, Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing 211166, China.
State Key Laboratory of Reproductive Medicine, Nanjing 211166, China.
Pathogens. 2022 Oct 26;11(11):1238. doi: 10.3390/pathogens11111238.
It is known that schistosome-derived antigens induce innate and adaptive immune responses that are essential for the formation of hepatic immunopathology. Here, we screened and synthesized four peptides derived from () heat shock protein 90α (Sjp90α-1, -2, -3, and -4), which is widely expressed in adults and eggs of the genus and induces remarkable immune reactions. To define the antigenicity of these peptides, we stimulated splenocytes with peptides, and the results showed that only the Sjp90α-1 peptide could predominately induce the activation of dendritic cells (DCs) and macrophages as well as alter the proportion of follicular helper T (Tfh) cells. Next, CD4 T cells were purified and cocultured with mouse bone-marrow-derived DCs (BMDCs) with or without Sjp90α-1 peptide stimulation , and the results showed that Sjp90α-1-stimulated BMDCs can significantly induce CD4 T-cell differentiation into Tfh cells, while the direct stimulation of CD4 T cells with Sjp90α-1 did not induce Tfh cells, indicating that the Sjp90α-1 peptide promotes Tfh cell differentiation depending on the presence of DCs. Furthermore, we selected and prepared an Sjp90α-1-peptide-based antibody and illustrated that it has excellent reactivity with the immunizing peptide and detects a single band of 29 kDa corresponding to the Sjp90α protein. The immunolocalization results showed that the protein recognized by this Sjp90α-1-peptide-based antibody is present in the mature eggs and the tegument of adults, implying that the parasite-derived peptide has a potential interaction with the host immune system. Finally, we evaluated antipeptide IgG antibodies and revealed a significantly higher level of anti-Sjp90α-1 peptide IgG antibodies in mice 3 weeks after infection. In conclusion, we illustrate that these synthetic peptides warrant further investigation by evaluating their antigen-specific immune response and their ability to efficiently induce Tfh cells. Moreover, they may constitute a potentially helpful method for the laboratory diagnosis of schistosomiasis japonica.
已知血吸虫衍生抗原可诱导先天性和适应性免疫反应,这些反应对于肝脏免疫病理学的形成至关重要。在此,我们筛选并合成了源自()热休克蛋白90α(Sjp90α-1、-2、-3和-4)的四种肽,该蛋白在该属的成虫和虫卵中广泛表达,并诱导显著的免疫反应。为了确定这些肽的抗原性,我们用肽刺激脾细胞,结果表明只有Sjp90α-1肽能主要诱导树突状细胞(DCs)和巨噬细胞的活化,并改变滤泡辅助性T(Tfh)细胞的比例。接下来,纯化CD4 T细胞并与有或无Sjp90α-1肽刺激的小鼠骨髓来源的DCs(BMDCs)共培养,结果表明Sjp90α-1刺激的BMDCs可显著诱导CD4 T细胞分化为Tfh细胞,而用Sjp90α-1直接刺激CD4 T细胞则不诱导Tfh细胞,这表明Sjp90α-1肽依赖DCs的存在促进Tfh细胞分化。此外,我们选择并制备了一种基于Sjp90α-1肽的抗体,并表明它与免疫肽具有优异的反应性,且检测到一条对应于Sjp90α蛋白的29 kDa单条带。免疫定位结果表明,这种基于Sjp90α-1肽的抗体识别的蛋白存在于成熟虫卵和成虫的体表,这意味着寄生虫衍生肽与宿主免疫系统存在潜在相互作用。最后,我们评估了抗肽IgG抗体,发现在感染后3周的小鼠中,抗Sjp90α-1肽IgG抗体水平显著更高。总之,我们表明这些合成肽通过评估其抗原特异性免疫反应及其有效诱导Tfh细胞的能力值得进一步研究。此外,它们可能构成一种潜在有用的日本血吸虫病实验室诊断方法。