Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Bone Marrow Mesenchymal Stem Cell-Derived Dermcidin-Containing Migrasomes enhance LC3-Associated Phagocytosis of Pulmonary Macrophages and Protect against Post-Stroke Pneumonia.

doi: 10.1002/advs.202206432

Figure Lengend Snippet: Figure 7. DCD is beneficial to AIS recovery and DCD-containing BM-MSC-derived migrasome effectively promotes phagocytosis of macrophages. A– G) Peripheral blood of AIS patients (acute phase, 0–3d after disease onset, n = 16) and healthy controls (HC, n = 8) were collected. (A) Plasma DCD concentration was assessed with ELISA. *p < 0.05, compared with HC by Student’s t-test (mean ± standard deviation). (B) Correlation of clinic parameters and plasma DCD concentration was assessed with Spearman correlation analysis and Point-biserial correlations. *p < 0.05. DM, diabetes mellitus, CHD, coronary heart disease. (C) Representative images of the magnetic resonance diffusion weighted imaging (MR-DWI) of AIS patients with low plasma DCD concentration (DCD ≤3.33 ng ml−1) or high plasma DCD concentration (DCD > 3.33 ng ml−1). (D) Association between plasma DCD concentration with infarct scale was estimated with Spearman correlation analysis. (E) Association between plasma DCD concentration with delta NIHSS (NIHSS at 7d minus NIHSS at 1d) was estimated with Spearman correlation analysis. (F) Representative images of the chest Computed Tomography (CT) of AIS patients with low plasma DCD concentration (DCD ≤3.39 ng ml−1, median of the cohort) or high plasma DCD concentration (DCD > 3.39 ng ml−1, median of the cohort). (G) Pie charts showing the occurrence of post-stroke pneumonia in AIS patients with low and high plasma DCD concentrations. H) DCD (1 ng ml−1), PBS-migrasomes (PBS-M, 50 μg ml−1) or E. Coli-migrasomes (E. Coli-M, 50 μg ml−1) labeled with Dil (red) were treated to BMDM (15 min). Immunostaining of WGA (green) and DCD (withe) in migrasome-treated BMDM was performed. Experiments were repeated for three times. I,J) BMDM were first pre-stimulated with DCD (1 ng ml−1), PBS-M (50 μg ml−1), or E. Coli-M (50 μg ml−1) for overnight then treated with E. Coli (E. Coli : BMDM = 20:1, 1 h). Phagocytic efficiency of BMDM to GFP expressing E. Coli was assessed with flow cytometry (I) and immunostaining (J). Experiments were repeated three times. **p < 0.01, compared with PBS-treated group by one-way ANOVA (mean ± standard deviation).

Article Snippet: Human Monocyte Enrichment and Macrophage Differentiation: Mononucleus cells were isolated from peripheral blood of healthy adults (age = 18–40y) with human peripheral blood monocyte isolation Solution kit (Solarbio, P8680).

Techniques: Derivative Assay, Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Imaging, Computed Tomography, Labeling, Immunostaining, Expressing, Cytometry

Journal: bioRxiv

Article Title: Stabilized gp120-specific CD4 for next-generation HIV-1 inhibitors

doi: 10.64898/2026.03.24.713825

Figure Lengend Snippet: a, Diagram of the flow cytometry experimental set-up. Mononuclear leukocytes were isolated by Lymphoprep gradient density separation, aliquoted, and stored frozen. Aliquots were thawed as needed and samples were prepared with live-dead staining and Human TruStain FcX Fc receptor blocking solution. The cells were then labeled with anti-CD19 conjugated to Brilliant Violet 650 and biotinylated CD4-Ig variants. After removing excess antibody, the cells were labeled with streptavidin conjugated to Alexa Fluor 488 and analyzed by flow cytometry. b-g, Flow cytometry plots of tonsillar cells. CD19 signal is plotted on the y-axis and streptavidin signal is plotted on the x-axis. The plots were gated on viable singlet cells, and the fluorescence thresholds were determined using the negative controls (unstained cells and stained cells labeled with unbiotinylated Avitagged CD4 YW -Ig). The proportion of double-positive cells for CD19 and CD4-Ig are indicated in the top right quadrant. h , Graphic of the tonsil organoid culture experimental set-up. Five to six million tonsil cells were aliquoted in 12-well transwell plates with each well containing a 1 mL outer reservoir of media. 20 μL from the outer reservoirs were sampled at 2 hours, 24 hours, 48 hours, 72 hours, and 96 hours post-transwell seeding. The outer reservoirs were mixed by pipetting prior to each collection and the timepoints were stored at -80 °C. i, j, Time-course measurements (left) of the concentration of CD4 YW -Ig and gCD4 YW -Ig incubated in tonsil culture media without tonsil cell seeding ( i ) or with tonsil cell seeding ( j ) and their area under the curves (right). Concentrations were determined by ELISA in technical triplicates using the respective proteins as standards. Each point is an average of three independent transwell samples and the error bars are s.d. Variants containing the Q40Y and T45W substitutions are denoted in subscript. CD4 YW -Ig for i and j additionally contain the thermostabilizing A55V, L109K, L151K, and L177E substitutions. gCD4 constructs for e, g, i , and j contain A55V, S60D, D63K, L109K, L151K, and L177E substitutions. Donor #664 was used for the data in b-g and donor #669 was used for the data in i and j .

Article Snippet: Samples were pelleted and resuspended in 50 μL of FACS buffer containing streptavidin-Alexa Fluor 488 conjugate (S32354; Invitrogen) diluted 1:50 (final concentration 40 μg/mL).

Techniques: Flow Cytometry, Isolation, Staining, Blocking Assay, Labeling, Fluorescence, Concentration Assay, Incubation, Enzyme-linked Immunosorbent Assay, Construct

Journal: Clinical and Experimental Immunology

Article Title: Anti‐vimentin/cardiolipin IgA in the anti‐phospholipid syndrome: A new tool for ‘seronegative’ diagnosis

doi: 10.1111/cei.13633

Figure Lengend Snippet: Prevalence of antibodies specific for vimentin/cardiolipin complex.

Article Snippet: Briefly, a 96‐well polystyrene plate (Thermo Fisher Scientific, Waltham, Massachusetts, USA) was coated with 100 μl/well of cardiolipin (50 μg/ml in methanol) (from bovine heart; Sigma‐Aldrich, St Louis, Missouri, USA) and human recombinant vimentin (5 μg/ml in 0.05 mM NaHCO 3 buffer, pH 9.5) (R&D Systems, Minneapolis, Minnesota, USA).

Techniques:

Journal: Clinical and Experimental Immunology

Article Title: Anti‐vimentin/cardiolipin IgA in the anti‐phospholipid syndrome: A new tool for ‘seronegative’ diagnosis

doi: 10.1111/cei.13633

Figure Lengend Snippet: Levels of anti‐vimentin/cardiolipin (aVim/CL) immunoglobulin (Ig)A in patients [anti‐phospholipid syndrome (APS), seronegative (SN)‐APS] and in healthy controls (HC). For detection of aVim/CL IgA all the sera were analyzed by enzyme‐linked immunosorbent assay (ELISA). The cut‐off level has been calculated as the 99th percentile of 40 HC sera

Article Snippet: Briefly, a 96‐well polystyrene plate (Thermo Fisher Scientific, Waltham, Massachusetts, USA) was coated with 100 μl/well of cardiolipin (50 μg/ml in methanol) (from bovine heart; Sigma‐Aldrich, St Louis, Missouri, USA) and human recombinant vimentin (5 μg/ml in 0.05 mM NaHCO 3 buffer, pH 9.5) (R&D Systems, Minneapolis, Minnesota, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Clinical and Experimental Immunology

Article Title: Anti‐vimentin/cardiolipin IgA in the anti‐phospholipid syndrome: A new tool for ‘seronegative’ diagnosis

doi: 10.1111/cei.13633

Figure Lengend Snippet: Distribution of positive seronegative‐anti‐phospholipid syndrome (SN‐APS) patients among those tested positive for at least one immunoglobulin (Ig)A assay: anti‐cardiolipin (aCL), anti‐β2‐glycoprotein I (aβ2‐GPI); anti‐vimentin/cardiolipin (aVim/CL)

Article Snippet: Briefly, a 96‐well polystyrene plate (Thermo Fisher Scientific, Waltham, Massachusetts, USA) was coated with 100 μl/well of cardiolipin (50 μg/ml in methanol) (from bovine heart; Sigma‐Aldrich, St Louis, Missouri, USA) and human recombinant vimentin (5 μg/ml in 0.05 mM NaHCO 3 buffer, pH 9.5) (R&D Systems, Minneapolis, Minnesota, USA).

Techniques:

Journal: Clinical and Experimental Immunology

Article Title: Anti‐vimentin/cardiolipin IgA in the anti‐phospholipid syndrome: A new tool for ‘seronegative’ diagnosis

doi: 10.1111/cei.13633

Figure Lengend Snippet: Receiver operating characteristic (ROC) analysis of seronegative‐anti‐phospholipid syndrome (SN‐APS). A Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI‐2K) variation presented an area under the curve (AUC) of 0.697 [95% confidence interval (CI) = 0.628–0.766, p < 0.0005]. The variation in anti‐vimentin/cardiolipin (aVim/CL) immunoglobulin (Ig)A levels presented an area under the curve (AUC) of 0.75 (95% CI = 0.645–0.856, p < 0.0001, Youden’s J = 0.213) for the development of thrombotic or pregnancy events, as indicated in the APS classification criteria

Article Snippet: Briefly, a 96‐well polystyrene plate (Thermo Fisher Scientific, Waltham, Massachusetts, USA) was coated with 100 μl/well of cardiolipin (50 μg/ml in methanol) (from bovine heart; Sigma‐Aldrich, St Louis, Missouri, USA) and human recombinant vimentin (5 μg/ml in 0.05 mM NaHCO 3 buffer, pH 9.5) (R&D Systems, Minneapolis, Minnesota, USA).

Techniques: Activity Assay