Journal: Viruses

Article Title: Interferon-λ Attenuates Rabies Virus Infection by Inducing Interferon-Stimulated Genes and Alleviating Neurological Inflammation

doi: 10.3390/v12040405

Figure Lengend Snippet: Treatment with IFN-λ inhibits RABV replication in vitro. NA or Vero cells were infected with B2c at a multiplicity of infection (MOI) of 0.01, and recombinant mouse IFN-λ2 or IFN-λ3 at 10 ng/mL and 1000 ng/mL was added to treat the B2c infected NA or Vero cells at 24 h post infection (hpi). Cell supernatants were harvested at 24 h and 48 h after the treatment for virus titration on NA cells ( A ) or Vero cells ( D ). The production of vRNA in NA cells ( B ) or Vero cells ( E) was measured by qRT-PCR. RABV N transcription levels in NA cells ( C ) or Vero cells ( F) were determined by qRT-PCR. Error bars represent the standard error (SE, n = 3). The following notations were used to indicate significant differences between groups: *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: At 24 hpi, the cells were treated with 10 or 1000 ng/mL of IFN-λ2 or IFN-λ3 (BD Biosciences, San Jose, CA, USA) by addition into the culture medium.

Techniques: In Vitro, Infection, Recombinant, Titration, Quantitative RT-PCR

Journal: Viruses

Article Title: Interferon-λ Attenuates Rabies Virus Infection by Inducing Interferon-Stimulated Genes and Alleviating Neurological Inflammation

doi: 10.3390/v12040405

Figure Lengend Snippet: Construction and characterization of recombinant RABVs expressing IFN-λ2 or IFN-λ3. ( A ) Schematic diagram describing the construction of B2c, rB2c-IFNλ2, and rB2c-IFNλ3. The pcDNA3.1-B2c plasmid was derived from CVS-B2c by deleting the long non-coding region of the G gene and adding BsiW I and Nhe I sites between the G and L genes. Murine IFN-λ2 or IFN-λ3 coding sequences were then inserted into the RABV genome between the G and L genes. ( B ) Expression of IFN-λ2 and IFN-λ3 was measured by by ELISA. Briefly, NA cells were infected with B2c, rB2c-IFNλ2, or rB2c-IFNλ3 (MOI = 1, 0.1, 0.01, or 0.001) for 24 h, and the cell culture supernatants were then harvested to determine the quantity of murine IFN-λ2 or IFN-λ3 using a commercial ELISA kit. ( C ) BSR, NA ( D ), or Vero cells ( E ) were infected with different rRABVs at a MOI of 0.01, and multiple-step growth curves were depicted according to the viral titers at different time points. Error bars represent the SE ( n = 3). ( F ) RABV N protein expression in NA cells after infection with rRABVs. N protein was detected by western blot at 48 hpi and ratios of N/β-actin were calculated using ImageJ. ( G ) Comparison of the fluorescence morphologies of NA cells infected by different rRABVs at 48 hpi. Twenty fluorescent foci were examined to determine the number of infected cells per fluorescent focus ( H ). Scale bars represent 200 μm. Error bars represented the SE ( n = 3). The following notations were used to indicate significant differences between groups: *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: At 24 hpi, the cells were treated with 10 or 1000 ng/mL of IFN-λ2 or IFN-λ3 (BD Biosciences, San Jose, CA, USA) by addition into the culture medium.

Techniques: Recombinant, Expressing, Plasmid Preparation, Derivative Assay, Enzyme-linked Immunosorbent Assay, Infection, Cell Culture, Western Blot, Fluorescence