Exploring the Impact of Adalimumab on Gene Expression in Retinal Pigment Epithelial Cells: Insights into Apoptosis, Inflammation, and Fibrogenesis
Fatemeh SanieJahromi1 *, M. Hossein Nowroozzadeh 1 , Mojtaba Yousefi 1 , Mostafa Abuali1
- Poostchi Ophthalmology Research Center, Department of Ophthalmology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Abstract: Adalimumab (ADA) is an anti-inflammatory antibody provisionally under investigation for the treatment of uveitis. This study aimed to assess the effect of ADA on apoptotic, inflammatory, and fibrogenesis gene expression at mRNA and protein levels in retinal pigment epithelial (RPE) cells.
Methods: RPEs were treated with serial concentrations of ADA (0.5x, x, 2x, and 4x; [x= 250 µg/mL]) for 24 hours. MTT assay was done and the mRNA and protein expressions were quantified using real-time PCR and ELISA assay, respectively.
Results: The mRNA levels of IL-1b and IL-6 were significantly increased in ADA-treated RPEs at 0.5x and x concentrations. However, the increase in cytokine secretion was observed only in IL-1b at x concentration. TGF-β was significantly upregulated in the 0.5x and 4x doses of ADA both at mRNA and protein levels. MTT assay also confirmed the safety of ADA on RPEs.
Conclusion: In conclusion, despite its safety, the 2x concentration of ADA was the only dose that did not ignite the gene expression of any of the studied inflammatory and fibrogenesis genes. This dosage, which is roughly equal to 2 mg intravitreal dose in a clinical setting, might be referred to as a reference starting point for future in-vivo studies in ocular conditions.
