Recombinant Human Interferon α-2b Nanoparticles in the Treatment of Human Papillomavirus Infection Model and Detection of LMX 1A Expression in Cervical Exhumation Cells

Abstract

Interferon (INF) is a kind of multifunctional and highly active protein-like cytokines produced by animal cells after stimulation, which exerts the broad-spectrum antiviral, anti-tumor, and immunomodulatory effects. Preparing the nanoparticles (NPs) loaded drug can improve the stability of protein polypeptides in vivo, prolong the biological half-life period, and improve the bioavailability. In this research, recombinant human interferon (RHUINF) α-2b NPs were prepared to explore their therapeutic effect on human papillomavirus (HPV) infection model. Firstly, the RHUINF α-2b poly (lactic acid-glycolic acid) copolymer (PLGA) of composite bioactive glass was prepared by electrostatic spray technology. The in vitro quality, stability, and pharmacokinetic characteristics in vivo of the NPs were measured. Then, a mouse model infected with HPV16 pseudovirus infection (NC group) was prepared. Next, they were treated by painting imiquimod (IMQ) (IMQ group), intramuscularly injecting the RHUINF α-2b (Intron A), intramuscularly injecting the RHUINF α-2b-Plga NP suspension (Intra/PLGA), and intramuscularly injecting the RHUINF α-2B-bioactive glass-PLGA NP suspension (Intron A/BAG/PLGA), respectively. In addition, the differences in serum inflammatory cytokines, vaginal biofluorescence activity (VBA), HPV load, and LMX 1A expressions in cervical exhumation cells (CECs) were analyzed. The average particle sizes (PSs) of Intron A/PLGA and Intron A/BAG/PLGA NPs were 352.3 nm and 385.1 nm, respectively, and no great differences were observed in encapsulation rate (ER), drug loading (DL), and in vivo release (IVR). The maximum peak times (MPTs) of Intron A, Intron A/PLGA, and Intron A/BAG/PLGA were 1.3 h, 6.2 h, and 6.5 h, respectively, and their maintenance durations were 5.5 h, 120.7 h, and 245.6 h, respectively. Based on the NC group, IFN-γ, IL-2, and TNF-α in the IMQ group, Intron A group, Intron A/PLGA group, Intron A/PLGA group, and Intron A/BAG/PLGA group were decreased, VBA and HPV load were decreased, and mRNA of LMX 1A in ECEs was increased (p < 0.05). Based on the Intron A group, the Intron A/PLGA and Intron A/BAG/PLGA groups exhibited decreased serum inflammatory factors (IFFs), VBA, HPV load, and LMX 1A mRNA in the CECs were increased (p < 0.05). By taking the Intron A/PLGA group as reference, the Intron A/BAG/PLGA group showed greatly reduced serum IFFs, VBA, HPV load, and LMX 1A mRNA expression in CECs (p<0.05). Preparing RHUINF α-2b NPs could prolong the half-life period of RHUINF α-2b in vivo, reduce the inflammatory response and load of HPV-DNA in HPV infection models, and upregulate LMX 1A in CECs. In addition, RHUINF α-2b NPs could improve the therapeutic effect of RHUINF α-2b.