Ultradiluted SARS-CoV-2 Spike Protein mitigates hyperinflammation in lung via ferritin and MMP-9 regulation in BALB/c mice

Virus Res. 2023 May:329:199091. doi: 10.1016/j.virusres.2023.199091. Epub 2023 Mar 15.

Abstract

Aim: 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.

Main methods: 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.

Key findings: 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.

Significance: 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.

Keywords: Ferritin; Gelatin zymography; Immunofluorescence; MMP-9; Spike protein of SARS-CoV-2; Ultradilution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COVID-19*
  • Cytokines / metabolism
  • Ferritins / genetics
  • Gelatin / metabolism
  • Humans
  • Inflammation
  • Lung / metabolism
  • Matrix Metalloproteinase 9 / analysis
  • Matrix Metalloproteinase 9 / genetics
  • Matrix Metalloproteinase 9 / metabolism
  • Mice
  • Mice, Inbred BALB C
  • SARS-CoV-2 / metabolism
  • Spike Glycoprotein, Coronavirus* / genetics
  • Spike Glycoprotein, Coronavirus* / metabolism
  • Tissue Inhibitor of Metalloproteinase-1 / metabolism

Substances

  • spike protein, SARS-CoV-2
  • Spike Glycoprotein, Coronavirus
  • Matrix Metalloproteinase 9
  • Tissue Inhibitor of Metalloproteinase-1
  • Ferritins
  • Gelatin
  • Cytokines

Supplementary concepts

  • SARS-CoV-2 variants