Journal: PLOS Neglected Tropical Diseases

Article Title: Development of novel monoclonal antibodies for detection of pan- Lassa virus

doi: 10.1371/journal.pntd.0014326

Figure Lengend Snippet: A) Cross-reactivity of mAbs to different LASV lineages NP was measured using cell lysate in indirect ELISA. OD 450 for each mAb is summarized as heat map. B) Representative of binding of anti-LASV NP mAb with cell lysate of HEK293T expressing LASV NPs (lineage I-VII) using WB, each lysate was diluted 1:100 times and used as an antigen for measurement of binding of mAb 28-2-3. C) Representative images of IFA showing binding of mAb 57-2 to the HEK293T cells expressing LASV NP, 24 hours post transfection cells were fixed, permeabilized and incubated with mAb 57-2 and detected by Alexa488 anti-mouse IgG antibody. Hoechst was used for nucleus staining. HEK293T cells transfected with pCAGGS empty vector were used as negative control (NC).

Article Snippet: The membrane was incubated with 1 μg/ml of mouse anti-LASV NP mAbs which were generated in house or commercial mAb from Zalgen (NP LASV MAb1474) was also used to detect LASV NP in the cell lysate.

Techniques: Indirect ELISA, Binding Assay, Expressing, Transfection, Incubation, Staining, Plasmid Preparation, Negative Control

Journal: PLOS Neglected Tropical Diseases

Article Title: Development of novel monoclonal antibodies for detection of pan- Lassa virus

doi: 10.1371/journal.pntd.0014326

Figure Lengend Snippet: The cross-competition profile of mAbs for binding to LASV NP was determined using Ab competitive ELISA. HEK293T cell lysate expressing LASV lineage III (Ojoko) NP was diluted to 1:100 in PBS and coated on the ELISA plate surface. Serially diluted unconjugated mAbs were used as primary antibody as represented in different colors in each graph and then 1µg/ml of HRP-conjugated mAbs for LASV NP were used as secondary antibody for each measurement. Anti-LASV GPC mAb was used as an IgG control.

Article Snippet: The membrane was incubated with 1 μg/ml of mouse anti-LASV NP mAbs which were generated in house or commercial mAb from Zalgen (NP LASV MAb1474) was also used to detect LASV NP in the cell lysate.

Techniques: Binding Assay, Competitive ELISA, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: PLOS Neglected Tropical Diseases

Article Title: Development of novel monoclonal antibodies for detection of pan- Lassa virus

doi: 10.1371/journal.pntd.0014326

Figure Lengend Snippet: A) 2x10 5 PFU/ml of LASV lineages I-IV were serially diluted and used as an antigen for sandwich ELISA with 1µg/ml of each capturing and detection mAbs. Binding of different pairs of capturing mAb/detection mAb with LASV NP are denoted in the graph as OD 450 . B) Sandwich ELISA for detection of LASV NP from lineages V-VII were measured using VLPs, lineage IV VLPs was used as positive control for this measurement. C) Measurement of NP mAbs binding with rVSV, EBOV, MARV, and LCMV at different concentration of authentic virus using different combinations of mAbs. Data represents means and standard deviations (SDs) of OD 450 from three biological triplicates. The threshold of sandwich ELISA was calculated as the mean + 3SDs of OD 450 value of negative control and represented as a black dotted horizontal line in the graphs.

Article Snippet: The membrane was incubated with 1 μg/ml of mouse anti-LASV NP mAbs which were generated in house or commercial mAb from Zalgen (NP LASV MAb1474) was also used to detect LASV NP in the cell lysate.

Techniques: Sandwich ELISA, Binding Assay, Positive Control, Concentration Assay, Virus, Negative Control

Journal: Frontiers in Immunology

Article Title: Immunization with MVA-based vaccines protects K18-hACE2 mice from SARS-CoV-2 infection-associated inflammatory lesions in brains

doi: 10.3389/fimmu.2026.1788665

Figure Lengend Snippet: Study design. Male and female K18-hACE2 mice received PBS, Modified Vaccinia Virus Ankara wild type (MVA-WT), recombinant MVA expressing native (S) or stabilized (ST) SARS-CoV-2 spike protein, nucleocapsid protein (N) or both ST and N proteins prior to intranasal SARS-CoV-2 infection (3.6 × 10 4 TCID50, Germany/BavPat1/2020 strain, NR-52370). (a) Booster experiment - mice were vaccinated twice (day 0 and 21) and infected four weeks later. (b) Prime experiment - mice were immunized only once and infected four weeks later. (c) Emergency experiment – animals were vaccinated once and infected two days later. Mice were sacrificed 6-, 7- and 8 days post infection (dpi). The syringe icon indicates vaccination, the virus icon denotes SARS-CoV-2 infection in the animals, and the red cross marks the day of euthanasia.

Article Snippet: The avidin-biotin-peroxidase complex method was implemented with the primary antibodies targeting anti SARS-CoV-2-NP (SARS-CoV-2 nucleocapsid protein, 40143-MM05, Sino Biological, Beijing, China) anti Iba-1 (ionized calcium-binding adapter molecule 1; #019-197, FUJIFILM Wako Pure Chemical Corporation, Richmond, USA; 1:8000) and anti CD3 (#A0452, Dako Agilent, Santa Clara, USA; 1:200) for SARS-CoV-2-NP, microglia/macrophages and T cells, respectively, as previously described ( ).

Techniques: Modification, Virus, Recombinant, Expressing, Infection

Journal: Frontiers in Immunology

Article Title: Immunization with MVA-based vaccines protects K18-hACE2 mice from SARS-CoV-2 infection-associated inflammatory lesions in brains

doi: 10.3389/fimmu.2026.1788665

Figure Lengend Snippet: In contrast to infected animals from both the PBS [ (a) 6 days post infection (dpi)] and MVA-WT [ (b) 6 dpi] groups, which showed lymphohistiocytic meningoencephalitis with microgliosis and neuroinvasion, K18-hACE2 mice, vaccinated twice [Booster experiment, (c) 8 dpi] or once [Prime experiment, (d) 8 dpi] four weeks prior to infection with MVA-based vaccines expressing stabilized spike and nucleocapsid proteins (MVA-SARS-2-ST/N) showed no inflammatory lesions and no viral spread. Animals immunized with MVA-SARS-2-ST/N two days prior to infection [Emergency experiment, (e) 6 dpi (two left pictures) and at 8 dpi (two right pictures)] also developed lesions, but the inflammatory alterations and the immunopositivity for the viral antigen were less pronounced than in the controls. (a, b) Control animals showed lymphohistiocytic meningoencephalitis, numerous SARS-CoV-2-NP-positive neurons, spiky microglia, and predominantly perivascular T cell infiltration. (c, d) Animals vaccinated twice (c) or once (d) four weeks prior to infection with MVA-SARS-2-ST/N presented no inflammatory changes or neuroinvasion. (e) Animals immunized (MVA-SARS-2-ST/N) once two days prior to infection presented less severe inflammatory changes and a less pronounced spread of viral antigen than the control groups. Bars = 50µm.

Article Snippet: The avidin-biotin-peroxidase complex method was implemented with the primary antibodies targeting anti SARS-CoV-2-NP (SARS-CoV-2 nucleocapsid protein, 40143-MM05, Sino Biological, Beijing, China) anti Iba-1 (ionized calcium-binding adapter molecule 1; #019-197, FUJIFILM Wako Pure Chemical Corporation, Richmond, USA; 1:8000) and anti CD3 (#A0452, Dako Agilent, Santa Clara, USA; 1:200) for SARS-CoV-2-NP, microglia/macrophages and T cells, respectively, as previously described ( ).

Techniques: Infection, Vaccines, Expressing, Control

Journal: Frontiers in Immunology

Article Title: Immunization with MVA-based vaccines protects K18-hACE2 mice from SARS-CoV-2 infection-associated inflammatory lesions in brains

doi: 10.3389/fimmu.2026.1788665

Figure Lengend Snippet: K18-hACE2 mice vaccinated once or twice four weeks prior to infection with MVA-based vaccines expressing native (S) or stabilized spike (ST) protein showed minimal or no inflammatory alterations, no presence of viral antigen or change in microglial morphology, and only minimal T cell infiltration in contrast to non-vaccinated animals from the PBS and MVA-WT groups and animals immunized with a MVA vaccine expressing the viral nucleocapsid (N) protein. Animals immunized with MVA-SARS-2-ST/N two days prior to infection also developed lesions, but the inflammatory changes and immunopositivity for viral antigen were less pronounced than in the control animals. (a-c) Semi-quantitative analysis of hematoxylin and eosin-stained and SARS-CoV-2-NP-immunostained slides as well as quantitative analysis of Iba-1-positive area and CD3-positive T cells in the cerebrum and brain stem of animals immunized with the Booster (a) , Prime (b) and Emergency (c) protocols. Data were tested using the Kruskal–Wallis test followed by Dunn–Bonferroni post hoc testing. Statistical significance was accepted at a p-value of ≤0.05 (*). The graphs show mean (solid line), individual values (dots), and standard deviation (vertical bars).

Article Snippet: The avidin-biotin-peroxidase complex method was implemented with the primary antibodies targeting anti SARS-CoV-2-NP (SARS-CoV-2 nucleocapsid protein, 40143-MM05, Sino Biological, Beijing, China) anti Iba-1 (ionized calcium-binding adapter molecule 1; #019-197, FUJIFILM Wako Pure Chemical Corporation, Richmond, USA; 1:8000) and anti CD3 (#A0452, Dako Agilent, Santa Clara, USA; 1:200) for SARS-CoV-2-NP, microglia/macrophages and T cells, respectively, as previously described ( ).

Techniques: Infection, Vaccines, Expressing, Control, Staining, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Immunization with MVA-based vaccines protects K18-hACE2 mice from SARS-CoV-2 infection-associated inflammatory lesions in brains

doi: 10.3389/fimmu.2026.1788665

Figure Lengend Snippet: In contrast to the animals from control groups (PBS, 6 dpi), which showed multiple neurons (predominantly in the ganglion cell layer) positive for SARS-CoV-2 nucleocapsid protein (NP) in the retina, mice vaccinated four weeks prior to the infection with MVA-SARS-2-ST/N (Booster experiment and Prime experiment, 8 dpi) did not show any viral antigen immunopositivity. Only one animal immunized with MVA-SARS-2-ST/N two days prior to the infection displayed SARS-CoV-2-NP positive neurons in the retina (Emergency experiment, 7 dpi). Abbreviations indicate the retinal layers: NFL, nerve fiber layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; PL, photoreceptor layer; Bars = 50µm.

Article Snippet: The avidin-biotin-peroxidase complex method was implemented with the primary antibodies targeting anti SARS-CoV-2-NP (SARS-CoV-2 nucleocapsid protein, 40143-MM05, Sino Biological, Beijing, China) anti Iba-1 (ionized calcium-binding adapter molecule 1; #019-197, FUJIFILM Wako Pure Chemical Corporation, Richmond, USA; 1:8000) and anti CD3 (#A0452, Dako Agilent, Santa Clara, USA; 1:200) for SARS-CoV-2-NP, microglia/macrophages and T cells, respectively, as previously described ( ).

Techniques: Control, Infection

Journal: bioRxiv

Article Title: c-Fos enhances influenza virus replication by stabilizing the M2 protein and promoting autophagosome accumulation

doi: 10.64898/2026.03.05.709812

Figure Lengend Snippet: (A) Cell viability was assessed by CCK-8 assay following T5224 treatment. For infection experiments, cells were pretreated with 0, 10, or 20 μM T5224 for 2 h before PR8 virus infection. Protein lysates were harvested at 24 h post-infection and analyzed by Western blot using anti-NP and anti-GAPDH antibodies. (B) 293H cells were co-transfection with M2 and c-Fos plasmids, alongside M2 and vector controls. After 48 h, cells were lysed and RNA was extracted for RT-qPCR analysis of M2 mRNA expression. (C and D) Co-immunoprecipitation and immunoblot analysis of 293H cells co-transfected with Flag-c-Fos and M2 (C) or NP (D). (E) 293H cells were co-transfection with Flag-c-Fos and M2 for 48 h, followed by immunostaining for Flag (red) and M2 (green). Representative confocal immunofluorescence images are shown. Scale bars, 2 μm. (F) 293H cells were co-transfected with M2 and Flag-c-Fos or empty vector. Cells were treated with 50 μg/mL CHX and harvested at the indicated time points (0 - 8 h). M2 protein levels were analyzed by Western blot. (G and H) 293H cells were transfected with M2 or empty vector. At 6 h post-transfection, cells were treated with MG132 (100, 200, 400 nM), Bafilomycin A1 (1, 2, 5 nM), or DMSO control. After 48 h, M2 protein expression was assessed by Western blot. (I) 293H cells were co-transfected with M2 and Flag-c-Fos or vector, followed by treatment with MG132 (200 nM) or Bafilomycin A1 (2 nM) for 48 h. M2 protein levels were analyzed by Western blot. All data are presented as means ± SD from at least three independent experiments. Significance was determined by Student’s t test: *, P < 0.05; **, P < 0.01; ***, P < 0.001. #, P > 0.05.

Article Snippet: The following primary antibodies were used: anti-LC3B (2775; Cell Signaling Technology, CST), anti-SQSTM1 (5114; CST), anti-GAPDH (2118; CST), anti-NP (10780-01; SouthernBiotech), anti-SERCA (A1097; ABclonal), anti-Flag (AE092; ABclonal), and anti-influenza A virus M2 (IT-003-015; Immune Technology).

Techniques: CCK-8 Assay, Infection, Virus, Western Blot, Cotransfection, Plasmid Preparation, Quantitative RT-PCR, Expressing, Immunoprecipitation, Transfection, Immunostaining, Immunofluorescence, Control