Effects of an intravitreal injection of interleukin-35-expressing plasmid on pro-inflammatory and anti-inflammatory cytokines

Int J Mol Med. 2016 Sep;38(3):713-20. doi: 10.3892/ijmm.2016.2688. Epub 2016 Jul 22.

Abstract

In order to explore the potential effects of interleukin (IL)-35 on IL-10, transforming growth factor-β (TGF-β), interferon-γ (INF)-γ, IL-12 and IL-17, a pcDNA3.1‑IL-35 plasmid was injected into the vitreous cavity of BALB/c mice. Enzyme-linked immunosorbent assay, western blot analysis and quantitative PCR analysis were performed to confirm the successful expression of IL-35. Slit-lamp biomicroscopy, hematoxylin and eosin staining and immunofluorescence were employed to detect the status of eyes, and western blot analysis was performed to examine the expression of corneal graft rejection-related cytokines. There were no abnormalities in the eyes pre-mydriasis or post-mydriasis and no injuries to the cornea or retina following the injection of IL-35-expressing plasmid. An immunofluorescence assay detected the positive expression of IL-35 in corneal epithelial cells from IL-35‑injected mice and negative staining in the control group. Further study revealed that IL-35 enhanced the expression of IL-10 and TGF-β which reached their highest levels at 1 and 2 weeks after injection, respectively (p<0.01). Moreover, the expression of INF-γ and IL-12 was decreased significantly at 2 weeks after the injection of IL-35-expressing plasmid (p<0.05), and the expression of IL-17 was suppressed notably at 4 weeks after the injection (p<0.05). The intravitreal injection of IL-35-expressing plasmid in mice downregulates the expression of pro-inflammatory cytokines and upregulates the expression of anti-inflammatory cytokines. Thus, IL-35 may further be assessed as a potential target for the treatment of corneal graft rejection.

MeSH terms

  • Animals
  • Blotting, Western
  • Cytokines / metabolism*
  • Female
  • Inflammation Mediators / metabolism*
  • Interferon-gamma / metabolism
  • Interleukin-10 / metabolism
  • Interleukin-12 / metabolism
  • Interleukin-12 Subunit p35 / genetics
  • Interleukin-12 Subunit p35 / metabolism
  • Interleukin-17 / metabolism
  • Interleukins / genetics
  • Interleukins / metabolism*
  • Intravitreal Injections
  • Mice, Inbred BALB C
  • Microscopy, Confocal
  • Minor Histocompatibility Antigens / genetics
  • Minor Histocompatibility Antigens / metabolism
  • Plasmids / administration & dosage
  • Plasmids / genetics
  • Receptors, Cytokine / genetics
  • Receptors, Cytokine / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transforming Growth Factor beta / metabolism
  • Vitreous Body / metabolism*

Substances

  • Cytokines
  • Ebi3 protein, mouse
  • Inflammation Mediators
  • Interleukin-12 Subunit p35
  • Interleukin-17
  • Interleukins
  • Minor Histocompatibility Antigens
  • Receptors, Cytokine
  • Transforming Growth Factor beta
  • interleukin-35, mouse
  • Interleukin-10
  • Interleukin-12
  • Interferon-gamma