Journal: Emerging Microbes & Infections

Article Title: Lactoferrin blocks orthopoxvirus entry via heparan sulphate and regulates host antiviral pathways

doi: 10.1080/22221751.2026.2631205

Figure Lengend Snippet: Lactoferrin inhibits orthopoxvirus infection by blocking viral entry through heparan sulphate binding. (A) Time-of-addition kinetics of 2.5 mg/ml bLf against vvTT infection (MOI = 0.1). Viral loads quantified at 14 hpi by Western blot and RT-qPCR. Data = mean ± SD ( n = 3). **** p < 0.0001 (one-way ANOVA with Dunnett's test). (B) Representative immunofluorescence images of vvTT-infected Vero E6 cells treated with 2.5 mg/ml bLf at indicated time points. Viral A27L protein (green), nuclei (DAPI, blue). Scale bars = 100 μm. (C) The inhibitory activity of lactoferrin at concentrations of 2.5, 1.25, and 0.625 mg/ml during the entry stage was quantified using RT-qPCR. Data = mean ± SD ( n = 3), **** p < 0.0001 (one-way ANOVA with Dunnett's test). (D) Post-entry curve depicted the replication dynamics of vvTT (MOI = 0.01) in Vero E6 cells after 2.5 mg/ml lactoferrin treatment. Tecovirimat at a concentration of 100 nM served as a positive control. Viral copies were quantified using a standard plasmid and analysed by absolute quantification methods. Data = mean ± SD ( n = 3). (E) Heparin competition assay. Lactoferrin (250 μg/ml) pre-incubated with heparin (HS) at indicated ratios during vvTT infection (MOI = 0.1). Viral loads quantified by RT-qPCR at 24 hpi. Data = mean ± SD ( n = 3). (F) Differential scanning fluorimetry showing direct lactoferrin-heparan sulphate binding. Thermal shift (Tm) of bLf (100 μg/ml) with increasing HS concentrations (0.02-200 μg/ml). Data = mean ± SD ( n = 3). (G) Sodium chlorate pre-treatment depletes cell-surface HSPGs and inhibit vvTT (MOI = 0.1) infection. Viral yields quantified by RT-qPCR at 24 hpi. Data = mean ± SD ( n = 3).

Article Snippet: Antiviral reference compounds brincidofovir (Cat. TQ0095, TargetMol) and tecovirimat (Cat. V3886, InvivoChem) were used for combination therapy studies.

Techniques: Infection, Blocking Assay, Binding Assay, Western Blot, Quantitative RT-PCR, Immunofluorescence, Activity Assay, Concentration Assay, Positive Control, Plasmid Preparation, Quantitative Proteomics, Competitive Binding Assay, Incubation

Journal: Emerging Microbes & Infections

Article Title: Lactoferrin blocks orthopoxvirus entry via heparan sulphate and regulates host antiviral pathways

doi: 10.1080/22221751.2026.2631205

Figure Lengend Snippet: Bovine lactoferrin exhibits therapeutic efficacy against VACV infection in vivo . (A) Experimental design for prophylactic bLf treatment. (B) Quantification of viral load in lung tissues. Viral genome copies were quantified as genomes per gram of tissue (left), while virus titres were calculated as PFU per gram of tissue (right). The data analysis performed using GraphPad Prism version 8.0. Data = mean ± SD ( n = 4), * p < 0.05, ** p < 0.01 (two-tailed t -test). (C) Quantitative analysis of infectious viral particles in the lung with 100 mg/kg lactoferrin, 50 mg/kg tecovirimat and PBS treatment based on plaque assay. (D) Weight loss dynamics and survival rates monitored daily for 7 days and clinical and pathological scoring of mice. (E) H&E staining and IHC analysis of the lung sections. The sections exhibited diffuse degeneration and necrosis of the epithelial lining (black arrows), along with haemorrhage, edema (red arrows), and fibrin exudation into the surrounding alveoli (blue arrows). IHC analysis conducted using a human anti-VACV A27L monoclonal antibody (brown signal). Mock was the blank control group without virus infection. The slides were digitized using a Pannoramic MIDI histoscanner (3DHISTECH), and the images analysed with CaseViewer software. Scale bars, each representing 50 micrometers, are provided for reference in each image.

Article Snippet: Antiviral reference compounds brincidofovir (Cat. TQ0095, TargetMol) and tecovirimat (Cat. V3886, InvivoChem) were used for combination therapy studies.

Techniques: Drug discovery, Infection, In Vivo, Virus, Two Tailed Test, Plaque Assay, Staining, Control, Software