Virology Laboratory, DAC Regional Research Institute, Kolkata, West Bengal 700035, India.
CCRH, Institutional Area, Janakpuri, New Delhi, Delhi 110058, India.
Virus Res. 2023 May;329:199091. doi: 10.1016/j.virusres.2023.199091. Epub 2023 Mar 15.
This study investigated the prophylactic and therapeutic role of ultradiluted preparation of the Delta variant of SARS-CoV-2 recombinant spike (S) protein during S antigen-induced inflammatory process of disease progression along with the probable mechanism of action.
Ultradiluted S protein (UDSP) was prepared and administered orally to adult BALB/c mice before and after administration of S antigen intranasally. After an observation period of 72 h, animals were sacrificed and expression level of ferritin was assayed through ELISA. The genetic expressions of cytokines, IL-6, IL-10, IL-1β, TNFα, IL-17, MMP-9, TIMP-1, ferritin light and heavy chains, and mitochondrial ferritin from lung tissues were investigated through RT-PCR. Formalin-fixed lung tissue sections were stained with hematoxylin and eosin to observe the degree of pathological changes. The activity of MMP-9 in lung tissues was investigated through gelatin zymography and immunofluorescence of MMP-9 in lung tissue sections was performed to revalidate the finding from gelatin zymography. Systems biology approach was used to elucidate a probable pathway where UDSP attenuated the inflammation through the regulation of pro- and anti-inflammatory cytokines.
UDSP attenuated the S antigen-induced hyperinflammation in the lung by regulating pro- and anti-inflammatory cytokines, calming cytokine storm, reducing ferritin level both in transcriptional and translational levels, and restoring critical ratio of MMP-9: TIMP-1.
Our findings suggest a probable pathway by which UDSP might have attenuated inflammation through the regulation of cytokines, receptors, and other molecules. This proclaims UDSP as a promising antiviral agent in the treatment of COVID-19-induced immunopathogenesis.
本研究旨在探讨超稀释的 SARS-CoV-2 德尔塔变异株重组刺突（S）蛋白在 S 抗原诱导疾病进展的炎症过程中的预防和治疗作用，以及可能的作用机制。
制备超稀释 S 蛋白（UDSP），并在鼻内给予 S 抗原前后口服给予成年 BALB/c 小鼠。观察 72 小时后，处死动物，通过 ELISA 测定铁蛋白表达水平。通过 RT-PCR 研究细胞因子、IL-6、IL-10、IL-1β、TNFα、IL-17、MMP-9、TIMP-1、肺组织中铁蛋白轻链和重链以及线粒体铁蛋白的基因表达。用苏木精和伊红染色福尔马林固定的肺组织切片，观察病理变化程度。通过明胶酶谱法研究肺组织中 MMP-9 的活性，并通过 MMP-9 在肺组织切片中的免疫荧光验证明胶酶谱法的发现。采用系统生物学方法阐明 UDSP 通过调节促炎和抗炎细胞因子来减轻炎症的可能途径。
UDSP 通过调节促炎和抗炎细胞因子、平息细胞因子风暴、降低转录和翻译水平的铁蛋白水平以及恢复 MMP-9:TIMP-1 的关键比值，减轻了 S 抗原诱导的肺部过度炎症。
我们的研究结果表明，UDSP 可能通过调节细胞因子、受体和其他分子来减轻炎症的可能途径。这表明 UDSP 有望成为治疗 COVID-19 引起的免疫发病机制的抗病毒药物。
