Journal: Journal of Biomedical Science

Article Title: Many but not all pathogen-associated molecular patterns aggravate neurogenic heterotopic ossification after spinal cord injury

doi: 10.1186/s12929-026-01237-y

Figure Lengend Snippet: Expression of PRRs by mouse and human skeletal muscle cells. Expression of PRR mRNA transcripts in mouse satellite cells (SC), FAPs (FP), endothelial cells (EC) and monocyte/macrophages (MΦ) sorted from skeletal muscles. Whole mouse bone marrow (BM) cells and splenocytes (SP) were used as positive controls for these transcripts. Human PDGFRα + FAPs were sorted from muscles surrounding NHO biopsies, and CD14 + monocytes were isolated from peripheral blood from healthy donors. A TLR1, B TLR2, C TLR3, D TLR4, E TLR5, F TLR6, G TLR7, H TLR8, I TLR9, J STING1, K RIGI, L MDA5 (IFIH1), M PRK, N NOD1, O NOD2, P Dectin-1 (CLEC7A), Q Dectin-2 (Clec4n/CLEC6A), R Mincle (CLEC4E). For each gene, left hand histogram represents expression of the mouse gene in mouse cells and the right-hand histogram represents expression of the human gene in human cells. For mouse cells, relative mRNA expression was quantified relative to house-keeping gene Hprt . For human cells, values were normalized using three references genes HPRT , RPLP0 and PPIA for FAPs and ACTB , GAPDH and PPIA for CD14 + cells. Each dot represents a mouse or a human donor, bars represent mean ± SD

Article Snippet: Human MPCs were trypsinized and incubated for 30 min with biotinylated anti-human PDGFRα (Cat# BAF322, R&D Systems) goat polyclonal antibody and CD56-PE (clone B159, BD Pharmigen) monoclonal antibody in PBS 2% FCS, 2 mM EDTA or with control isotypes IgG1 PE (Cat# A07796, Beckman Coulter) and biotinylated goat IgG (Cat# BAF108, R&D Systems).

Techniques: Expressing, Muscles, Isolation

Journal: Journal of Biomedical Science

Article Title: Many but not all pathogen-associated molecular patterns aggravate neurogenic heterotopic ossification after spinal cord injury

doi: 10.1186/s12929-026-01237-y

Figure Lengend Snippet: OSM and IL-1 neutralization in monocyte-conditioned media strongly inhibit hFAPs mineralization. A OSM, IL-1α and IL-1β concentrations quantified in conditioned media from human CD14 + macrophages stimulated with 200 ng/ml Pam2CSK4 (CM Pam2CSK4 ), 200 ng/ml Pam3CSK4 (CM Pam3CSK4 ) or non-stimulated (CM ∅ ). B OSM, IL-1α and IL-1β concentrations in CM Pam2CSK4 , CM Pam3CSK4 and CM ∅ were correlated with hFAP calcium mineralization measured by Alizarin Red staining after 2 weeks of culture in osteogenic conditions in the presence of the conditioned media. Each dot represents a different conditioned medium sample. C–F Mouse anti-human OSM antibody, isotype control antibody and IL-1RA were used to neutralize OSM and IL-1 in human CD14 + monocyte conditioned media as indicated below each chart. Calcium mineralization of hFAPs cultured for 2 weeks in osteogenic conditions with C CM Pam2CSK4 or D CM Pam3CSK4 was measured using Alizarin Red staining and quantified by spectrophotometry. E, F RUNX2 protein quantification by Western blot using Stain-Free normalization for hFAPs cultured for 2 weeks in osteogenic conditions with E CM Pam2CSK4 or F CM Pam3CSK4 . In C-F, each dot represents a hFAP individual donor. Bars represent mean ± SD, One-way ANOVA Dunnett’s multiple comparison test versus CM ∅ in ( A ) and ( B ), versus CM Pam2CSK4 in ( C ) and ( E ), or versus CM Pam3CSK4 in ( D ) and ( F ), * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Human MPCs were trypsinized and incubated for 30 min with biotinylated anti-human PDGFRα (Cat# BAF322, R&D Systems) goat polyclonal antibody and CD56-PE (clone B159, BD Pharmigen) monoclonal antibody in PBS 2% FCS, 2 mM EDTA or with control isotypes IgG1 PE (Cat# A07796, Beckman Coulter) and biotinylated goat IgG (Cat# BAF108, R&D Systems).

Techniques: Neutralization, Staining, Control, Cell Culture, Spectrophotometry, Western Blot, Comparison

Journal: EBioMedicine

Article Title: Quantification, epitope mapping and genotype cross-reactivity of hepatitis B preS-specific antibodies in subjects vaccinated with different dosage regimens of BM32

doi: 10.1016/j.ebiom.2020.102953

Figure Lengend Snippet: Comparison of preS-specific isotype and IgG subclasses responses in the four groups of patients. Shown are optical density (OD) values ( y -axes) corresponding to preS-specific IgG, IgA, IgM, IgE and IgG subclass (IgG 1 –IgG 4 ) levels in patients immunized with 3, 4 or 5 injections of BM32 or with placebo (see inlay) at baseline (V3), after one (V8) or four (V11) months treatment ( x -axes). Medians (horizontal bars) and significant differences are indicated: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The detection was performed using either biotin-labeled mouse anti-human IgG 1 (BD, Cat. 555869) or biotin-labeled mouse anti-human IgG 4 (BD, Cat. 555882) in dilution 1:1000 mixed with HRP-conjugated streptavidin (BD, Cat. 554066) in dilution 1:2500 and incubated overnight at 4 °C.

Techniques:

Journal: EBioMedicine

Article Title: Quantification, epitope mapping and genotype cross-reactivity of hepatitis B preS-specific antibodies in subjects vaccinated with different dosage regimens of BM32

doi: 10.1016/j.ebiom.2020.102953

Figure Lengend Snippet: Quantification of IgG 1 and IgG 4 subclass responses in patients treated with placebo or different numbers of BM32 injections. Shown are absolute increases of serum concentrations of IgG 1 and IgG 4 ( y -axes: μg/ml) specific (a) for preS, (b) for peptide A, (c) for peptide B and (d) for peptide C from baseline (V3) to visits 8 and 11 ( x -axes) inpatients immunized with placebo, 3, 4 or 5 injections of BM32. Medians (horizontal bars) and significant differences are indicated: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The detection was performed using either biotin-labeled mouse anti-human IgG 1 (BD, Cat. 555869) or biotin-labeled mouse anti-human IgG 4 (BD, Cat. 555882) in dilution 1:1000 mixed with HRP-conjugated streptavidin (BD, Cat. 554066) in dilution 1:2500 and incubated overnight at 4 °C.

Techniques:

Journal: EBioMedicine

Article Title: Quantification, epitope mapping and genotype cross-reactivity of hepatitis B preS-specific antibodies in subjects vaccinated with different dosage regimens of BM32

doi: 10.1016/j.ebiom.2020.102953

Figure Lengend Snippet: Kinetics and fine specificities of anti-preS antibody responses in the individual subjects of the placebo group, and the groups having received 3, 4 or 5 injections. The heat maps depict the log 10 of the serum concentrations of preS- or peptide-specific (a) IgG 1 and (b) IgG 4 . Missing values are indicated in gray.

Article Snippet: The detection was performed using either biotin-labeled mouse anti-human IgG 1 (BD, Cat. 555869) or biotin-labeled mouse anti-human IgG 4 (BD, Cat. 555882) in dilution 1:1000 mixed with HRP-conjugated streptavidin (BD, Cat. 554066) in dilution 1:2500 and incubated overnight at 4 °C.

Techniques: