CD40/CD40 LIGAND INTERACTIONS AND TNFα TREATMENT REDUCE ACTIVITY OF P105 PROMOTER OF THE HUMAN PAPILLOMA VIRUS-18 IN VITRO

Abstract

Background: Cervical carcinoma cells including those infected with the oncogenic human papilloma virus (HPV) and several cervical carcinoma cell lines show a strong expression of the CD40 receptor, unlike benign cervical epithelial cells infected with HPV. The functional relevance of this up-regulated expression in the tumor is not fully understood. Nevertheless, it might offer a unique possibility to target those malignant cells due to the antiviral and antitumoral effects of the CD40/CD40 ligand (CD40L) interactions. Aim: In vitro assessment of the effect of CD40L on HPV 18-P105 promoter activity and the subsequent release of IL-6 by the promoter transfected HeLaCD40 cells, which express CD40 constitutively. Material and Methods: Transfection of HeLaCD40 cells was achieved by electroporation after optimizing the parameters by the pCMV-β-Gal vector and β-Gal stain. Transfected HeLaCD40 cells were challenged with BHKCD40L and TNFα, in addition to BHKwt and medium alone as controls. HPV18P105 promoter activity was demonstrated by luciferase reporter gene assay while IL-6 was assessed by ELISA. Results: CD40/ CD40L interactions and TNFα treatment significantly reduced HPV18-P105 promoter activity (56.0 ± 10.2% and 64.1 ± 9.1% vs. control, respectively; p < 0.001). Likewise, IL-6, which is a sensitive cytokine of CD40 activation, was significantly increased in HeLaCD40 cells in the same experiments (2.7 fold after stimulation with BHKCD40L and 5.2 fold after stimulation with TNFα vs. control; p < 0.01 and p < 0.001, respectively). Conclusion: It is likely that the CD40/CD40L interactions and TNFα are effective against cervical carcinomas by repressing transcriptional activity of HPV promoter. This can result in new adjuvant treatments.

Activities of stromal and immune cells in HPV-related cancers

“…CD40/CD40L interaction is a central signalling pathway for dendritic maturation and for inducing CD4 + and CD8 + responses during HPV chronic infection [ 67 ]. In a HeLa model of cervical carcinoma study, , this interaction was able to repress transcriptional activity of HVP18-P105 promoter and increase IL-6 synthesis [ 68 ]. Thus, therapeutic approaches were developed/tested (e.g.…”

“…clinicaltrials.gov identifier NCT00319748) or are under clinical trials (e.g. NCT03418480) to induce the activation of this pathway aiming the prevention of hrHPV genes expression and promotion of higher CD4 + and CD8 + responses [ 68 , 69 ].…”

Impact of CD40 gene polymorphisms on the risk of cervical squamous cell carcinoma: a case-control study

“…These findings were further confirmed by Huang Q et al [ 29 ]. In vivo studies have shown that the expression of CD40 is correlated with HPV positivity, vascular endothelial growth factor (VEGF) expression, and microvessel density [ 27 , 30 ]. In recent years, various studies have shown that CD40 gene polymorphisms are related to some malignant tumors, such as non-Hodgkin’s lymphoma [ 31 ], breast cancer [ 32 ], and lung cancer [ 33 , 34 ].…”