Journal: Journal of Immunology Research

Article Title: Pdcd1 −/− Mice Exhibit High Susceptibility to Respiratory Syncytial Virus Infection

doi: 10.1155/jimr/1223280

Figure Lengend Snippet: Pdcd1 −/− mice are susceptible to intranasal RSV infection. Mice were inoculated intranasally with 10 6 PFU of purified RSV A2 strain. (A) Body weight was monitored daily for 7 dpi. At the time of inoculation (0 dpi), the body weights of mice were set as the baseline, and subsequent weight changes relative to the baseline are shown ( n = 3–5 mice per group). (B) Viral loads in the lungs of RSV‐infected mice were measured at the indicated time points. n = 5 mice per group. (C) Lungs were collected on 0, 4, and 7 dpi and subjected to H&E staining. Scale bars, 50 μm. (D) Statistical analysis based on histological scores for perivascular and alveolar wall inflammatory cell infiltration. n = 5 mice per group. All data are presented as mean ± SD. Statistical significance was determined by two‐way ANOVA with Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001 versus WT mice; ns, not significant.

Article Snippet: RSV strains long (ATCC VR‐26) and A2 (ATCC VR‐1540) were used for mouse challenge experiments.

Techniques: Infection, Purification, Staining, Analysis

Journal: bioRxiv

Article Title: The actin-binding protein drebrin interacts with the respiratory syncytial virus matrix protein and promotes virus particle release

doi: 10.64898/2026.10.05.756730

Figure Lengend Snippet: (A) HEp-2 cells were transfected with control (si NT) or drebrin siRNAs (si drebrin) for 48h and subsequently infected with RSV-WT (A,D) or RSV-mCherry (B-C) at MOI 1 for 24 h. (A) Viral titers were determined by plaque assay and normalized to the si NT condition. Bar graph shows mean ± SD from 3 independent experiments performed in triplicate. A one-sample t-test was performed. ns : P > 0.05. (B) Graph of mCherry signal at different time post-infection in si NT (black) and si drebrin (red) infected cells. The data shown are the mean ± SD of 3 independent experiments performed in triplicate. (C) mCherry intensity in si drebrin cells at 24 h post-infection relative to si NT. A one sample t-test was performed, ns : P > 0.05. (D) Cells were stained with an antibody against drebrin (red in merge) and G-RSV (green in merge) and with phalloidin (blue in merge). Representative images from 3 independent experiments are shown. One representative Z step is shown. Scale bar 10 μm.

Article Snippet: The recombinant wild-type RSV and RSV-mCherry viruses are derivated from the human RSV Long strain (ATCC VR-26) and were previously described ( , ).

Techniques: Transfection, Control, Infection, Plaque Assay, Staining

Journal: bioRxiv

Article Title: The actin-binding protein drebrin interacts with the respiratory syncytial virus matrix protein and promotes virus particle release

doi: 10.64898/2026.10.05.756730

Figure Lengend Snippet: Drebrin promotes viral particle release . (A) Schematic representation of the release assay. HEp-2 cells were transfected with control or specific siRNAs and seeded onto Transwell inserts for 48h (step 1). Cells were then infected with RSV-mCherry at an MOI of 1 (step 2). After washes to remove non-internalized virus, the infected “donor” cells were then placed in contact with target cells (step 3). The number of infected “donor” and “target” cells were quantified based on mCherry fluorescence using the Incucyte S3 system at 20h (step4) and 38h (step 5) post-infection respectively. RSV release efficiency was calculated as the ratio of infected target cells to infected donor cells and normalized to the control siRNA condition. (B) HEp-2 cells were transfected with si NT or drebrin-specific siRNAs (si drebrin) and release assay was performed as described in A. Date are mean ± SD of 6 independent experiments, each point represents mean of triplicates. Y axis represents viral release efficiency (ratio of infected target cells to infected donor cells) normalized to the control siRNA condition. (C) release assay was performed with HEp-2-YFP and HEp-2-Drebrin-YFP cells. Data are mean ± SD of 4 independent experiments done in triplicates. Y axis represents viral release efficiency (ratio of infected target cells to infected donor cells) normalized to the control siRNA condition. (B-C) A one-sample t-test was performed. * : P < 0.05; ** : P < 0.001.

Article Snippet: The recombinant wild-type RSV and RSV-mCherry viruses are derivated from the human RSV Long strain (ATCC VR-26) and were previously described ( , ).

Techniques: Release Assay, Transfection, Control, Infection, Virus, Fluorescence

Journal: bioRxiv

Article Title: The actin-binding protein drebrin interacts with the respiratory syncytial virus matrix protein and promotes virus particle release

doi: 10.64898/2026.10.05.756730

Figure Lengend Snippet: (A) Drebrin-YFP HEp-2 cells were uninfected or infected with RSV at MOI 1 for 24 h. Proteins associated with drebrin-YFP were affinity-purified using GFP nanobody-coupled agarose beads, identified by mass spectrometry, and quantified based on spectral counts obtained for each sample. A log2 fold change was calculated by comparing RSV-infected and uninfected conditions. Only proteins with at least two spectral counts in at least two biological replicates in either condition were included in the analysis. Statistical significance was assessed using a beta-binomial test adapted for spectral count data. Results are presented as a volcano plot. The horizontal dashed gray line indicates the statistical significance threshold ( P < 0.05), whereas the vertical dashed gray lines indicate a twofold difference between infected and uninfected samples. Cellular proteins used in C are indicated in blue. Viral RSV proteins are indicated in black. Results from three independent biological replicates (B) Left: Heat map of RT-qPCR analysis comparing mRNA levels of 17 candidate proteins in RSV-infected and uninfected cells. Blue indicates > 3-fold upregulation and yellow < 3-fold. Right: Heat map of viral replication analysis comparing RSV-mCherry signal in RSV-infected cells treated with interactors-specific siRNA and si NT. Blue indicates replication >140%, brown replication < 60% and yellow 60% < replication >140%. Complete data are shown in Fig S8. (C) HEp-2 cells were depleted with the indicated siRNAs and viral release assay was performed and compared with control conditions. One-Way ANOVA with Dunnett’s multiple comparisons test and Geisser-Greenhouse correction was performed relative to si NT. *** : P <0.001, ns : P >0,05.

Article Snippet: The recombinant wild-type RSV and RSV-mCherry viruses are derivated from the human RSV Long strain (ATCC VR-26) and were previously described ( , ).

Techniques: Infection, Affinity Purification, Mass Spectrometry, Analysis, Quantitative RT-PCR, Release Assay, Control, Significance Assay