influenza b virus ibv victoria  (ATCC)

Journal: Cell Communication and Signaling : CCS

Article Title: Induction of antiviral RNA interference by avian coronavirus IBV in PBMCs-Mφ

doi: 10.1186/s12964-026-02656-y

Figure Lengend Snippet: Phenotypic differences between QX and M41 in SPF chickens. A Pattern diagram of related phenotype detection in SPF chickens infected with IBV QX and IBV M41. B-C qRT–PCR was used to assess relative IBV-N mRNA expression levels in the trachea, lung, proventriculus, and kidney after infection with IBV-QX ( B ) and IBV-M41 ( C ) at the 24 and 48 h time points. Data show expression calculated relative to that of β-actin and are presented as the fold changes relative to that of the blank group (an equal amount of PBS was added as a control), normalized to 1. D IBV QX and M41 loads in the trachea, lung, proventriculus, and kidney detected by IHC at the 48 h infection time point (original magnification: 40×). The data are shown as the mean ± SD; n = 3. The differences were not significant at P ≥ 0.05 (ns) or significant at P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***)

Article Snippet: Afterward, PBMCs-Mφ and tissue sections were permeabilized with 0.4% Triton X-100 for 15 min. After being washed with PBS, the PBMCs-Mφ were stained with anti-IBV N antibody (HyTest, 1:500) in 5% bovine serum albumin for 1 h and were incubated with Alexa Fluor 594-labeled secondary antibodies (Bioss, 1:1,000) for 2 h and mounted with the SlowFadeTM Gold Antifade Mountant with DAPI (Invitrogen).

Techniques: Infection, Quantitative RT-PCR, Expressing, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Induction of antiviral RNA interference by avian coronavirus IBV in PBMCs-Mφ

doi: 10.1186/s12964-026-02656-y

Figure Lengend Snippet: QX and M41 induce different RNAi phenomena in PBMCs-Mφ. A-B Viral loads in PBMCs, red cells, and other cells were determined using qRT–PCR after IBV-QX ( A ) and IBV-M41 ( B ) infection for 24 and 48 h. C Microscopic pictures of PBMCs-Mφ on the indicated culture days (Days 1–3), and the black arrows indicate PBMCs-Mφ. Original magnification scale bar, 50 μm. D FCM results of macrophage sorting based on the PE-KUL01 antibody from the peripheral blood of chickens. E MHC II expression levels in PBMCs-Mφ were detected by flow cytometry after infection with IBV-QX and IBV-M41 at 24 and 48 h. The mean fluorescence intensity (MFI percentage) of MHC II was quantified, as shown on the grey side. F Confocal microscopy was used to examine PBMCs-Mφ infected with IBV-QX and IBV-M41 at 24 and 48 h. The cells were labelled with DAPI (blue) and an IBV N protein antibody (red). Scale bar, 10 μm. G Fluorescence intensity analysis of the data in Fig. 2F. H Viral titres in IBV QX- and M41-infected PBMCs-Mφ from 12–60 h were detected using the TCID 50 method; n = 3 per group. The data are shown as the mean ± SD; n = 3. The differences were not significant at P ≥ 0.05 (ns) or significant at P < 0.05 (*) or P < 0.01 (**)

Article Snippet: Afterward, PBMCs-Mφ and tissue sections were permeabilized with 0.4% Triton X-100 for 15 min. After being washed with PBS, the PBMCs-Mφ were stained with anti-IBV N antibody (HyTest, 1:500) in 5% bovine serum albumin for 1 h and were incubated with Alexa Fluor 594-labeled secondary antibodies (Bioss, 1:1,000) for 2 h and mounted with the SlowFadeTM Gold Antifade Mountant with DAPI (Invitrogen).

Techniques: Quantitative RT-PCR, Infection, Expressing, Flow Cytometry, Fluorescence, Confocal Microscopy

Journal: Cell Communication and Signaling : CCS

Article Title: Induction of antiviral RNA interference by avian coronavirus IBV in PBMCs-Mφ

doi: 10.1186/s12964-026-02656-y

Figure Lengend Snippet: A Relative mRNA expressions of IBV-N after IBV (QX and M41) infected BMDCs. B Statistical analysis of vsiRNAs after 24 h of IBV infection in BMDCs. C Visualization of the 21-23 nt vsiRNAs distribution and the enrichment region in the genome.

Article Snippet: Afterward, PBMCs-Mφ and tissue sections were permeabilized with 0.4% Triton X-100 for 15 min. After being washed with PBS, the PBMCs-Mφ were stained with anti-IBV N antibody (HyTest, 1:500) in 5% bovine serum albumin for 1 h and were incubated with Alexa Fluor 594-labeled secondary antibodies (Bioss, 1:1,000) for 2 h and mounted with the SlowFadeTM Gold Antifade Mountant with DAPI (Invitrogen).

Techniques: Infection

Journal: Cell Communication and Signaling : CCS

Article Title: Induction of antiviral RNA interference by avian coronavirus IBV in PBMCs-Mφ

doi: 10.1186/s12964-026-02656-y

Figure Lengend Snippet: PS-vsiRNA construction and antiviral validation. A - B Map of psilencer4.1-vsiRNA (PS-vsiRNA) plasmid construction ( A ) and identification of Bam HI/ Hin dIII enzyme digestion ( B ), including PS-Q10, PS-Q16, PS-M10, and PS-M16. C-E PBMCs-Mφ were transfected with PS-vsiRNA plasmids (PS-Q10, PS-Q16, PS-M10, and PS-M16) for 24 h and subjected to infection with IBV-QX and IBV-M41 ( C ). qRT–PCR was used to detect Dicer, Ago2 ( D ), and IBV-N ( E ) mRNA expression. Notably, the M41 group (PS-M10 and PS-M16) and the QX group (PS-Q10 and PS-Q16) were infected with IBV (QX and M41). F western blotting was used to analyse the expression levels of β-actin, IBV-N, and Dicer in the IBV-infected PBMCs-Mφ group. The right panel shows the grayscale analysis results. The data are shown as the mean ± SD; n = 3. The differences were not significant at P ≥ 0.05 (ns) or significant at P < 0.05 (*), P < 0.01 (**), P < 0.001 (***), or P < 0.0001 (****)

Article Snippet: Afterward, PBMCs-Mφ and tissue sections were permeabilized with 0.4% Triton X-100 for 15 min. After being washed with PBS, the PBMCs-Mφ were stained with anti-IBV N antibody (HyTest, 1:500) in 5% bovine serum albumin for 1 h and were incubated with Alexa Fluor 594-labeled secondary antibodies (Bioss, 1:1,000) for 2 h and mounted with the SlowFadeTM Gold Antifade Mountant with DAPI (Invitrogen).

Techniques: Biomarker Discovery, Plasmid Preparation, Transfection, Infection, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Induction of antiviral RNA interference by avian coronavirus IBV in PBMCs-Mφ

doi: 10.1186/s12964-026-02656-y

Figure Lengend Snippet: The QX-NSP10/NSP16 complex is a potential VSR protein. A FCM was used to analyse the changes in MHC II expression after transfection with PC-Q10, PC-Q16, PC-M10, or PC-M16. MFI shows the intensity of MHC II, where CpG and LPS are positive controls and PC is a negative control. B - D The relative mRNA expression levels of IBV-N ( B ), IFN-γ ( C ), Dicer, and Ago2 ( D ) were assessed by qRT–PCR after PBMCs-Mφ were transfected with PC, PC-M10 and PC-M16 (the M41 group), or PC-Q10 and PC-Q16 (the QX group) and infected with IBV (QX and M41). The data are shown as the means ± SDs; N = 3. The difference value analysis results are as follows: **** P < 0.0001 , *** P < 0.001 , ** P < 0.01 , * P < 0.05 , ns P > 0.05

Article Snippet: Afterward, PBMCs-Mφ and tissue sections were permeabilized with 0.4% Triton X-100 for 15 min. After being washed with PBS, the PBMCs-Mφ were stained with anti-IBV N antibody (HyTest, 1:500) in 5% bovine serum albumin for 1 h and were incubated with Alexa Fluor 594-labeled secondary antibodies (Bioss, 1:1,000) for 2 h and mounted with the SlowFadeTM Gold Antifade Mountant with DAPI (Invitrogen).

Techniques: Expressing, Transfection, Negative Control, Quantitative RT-PCR, Infection