Journal: Kidney International Reports

Article Title: Early-Stage B-cells Predict Relapse After Rituximab Treatment in Patients With Membranous Nephropathy

doi: 10.1016/j.ekir.2026.106365

Figure Lengend Snippet: Reconstitution of B-cells over time. To characterize B-cell subsets, peripheral blood mononuclear cells were stained with fluorochrome-conjugated monoclonal antibodies (BD Biosciences) directed against the following antigens: CD45 (V500-C), CD3 (APC), CD19 (APC-H7), CD27 (BV421), IgD (PE), CD38 (BV711), CD4 (BV605), and CD8 (PE), as well as 7-AAD (Miltenyi Biotech). The stained cells were then analyzed using multicolor flow cytometry (BD FACS Lyric). The subsets of gated CD19 + cells were identified based on surface marker expression as follows: naïve (CD19+CD27-IgD+), nonswitched memory (CD19+CD27+IgD+), switched memory (CD19+CD27+IgD-), and double negative (CD19+CD27-IgD-); and within the CD19+CD38++ population, we distinguished transitional cells (CD19+CD38++CD27-IgD+), plasmablasts (CD19+CD38++CD27+IgD-), and double-negative CD38 + cells (CD19+CD38++CD27-IgD-). B-cell subsets were expressed as a percentage of the total lymphocyte count. (a) B-cell subpopulations were assessed in 39 patients at baseline and in 36 patients at 3 (M3), 6 (M6), 9 (M9), 12 (M12), and 18 (M18) months following the first rituximab infusion. The 3 patients who did not receive treatment were excluded from the follow-up. Complete depletion of B-cells was observed in all patients at M3 post-rituximab for all B-cell subpopulations. CD19 + cells reappeared 6 months after rituximab infusion. Naive cells re-emerged the most among B-cells, followed by CD38 + cells, transitional cells and finally memory cells. Data are shown as mean values (dots). (b–k) B-cell subpopulations at baseline and at subsequent time points were compared between relapsing patients ( n = 8) and nonrelapsing patients ( n = 19); the 2 patients who received additional anti-CD20 infusions were excluded from subsequent analyses. Data are shown as medians and interquartile range (IQR). P -values were calculated by comparing the median values of each cell subpopulation between relapsing and nonrelapsing patients using a nonparametric, unpaired Mann–Whitney U test.

Article Snippet: To characterize B-cell subsets, peripheral blood mononuclear cells were stained with fluorochrome-conjugated monoclonal antibodies (BD Biosciences) directed against the following antigens: CD45 (V500-C), CD3 (APC), CD19 (APC-H7), CD27 (BV421), IgD (PE), CD38 (BV711), CD4 (BV605), and CD8 (PE), as well as 7-AAD (Miltenyi Biotech).

Techniques: Staining, Bioprocessing, Flow Cytometry, Marker, Expressing, MANN-WHITNEY

Journal: Materials Today Bio

Article Title: Injectable antifibrotic drug-loaded hydrogels reduce fibrosis and restore myogenesis by enhancing mitochondrial metabolism and cell mechanics in an in vitro coculture model

doi: 10.1016/j.mtbio.2026.103033

Figure Lengend Snippet: Establishment and characterization of a fibrotic macrophage-muscle fibrosis model. a) Brightfield image of unstimulated RAW264.7 cells in the top chamber of a RAW-C2C12 coculture system, b) Brightfield image of C2C12 cells in the bottom chamber under basal coculture conditions, c) Fluorescence image of C2C12 cells in the bottom chamber exhibiting myotube morphology, d) Fluorescence image confirming the absence of fibrotic marker expression in C2C12 cells under basal conditions, e) Brightfield image of LPS-activated RAW264.7 cells in the top chamber, f) Brightfield image of C2C12 cells in the bottom chamber displaying a myofibroblast-like morphology post-LPS activation, g) Fluorescence image of phenotypic transition of C2C12 cells to myofibroblast-like cells, h) Fluorescence image demonstrating fibrotic protein expression in LPS-treated C2C12 cells (magenta arrow indicates CD38 cells). Immunostaining images: In panels c) and g), TGF-β, DAPI, and actin are colored red, blue, and green, respectively. In panels d) and h), COL1, α-SMA, CD38, and DAPI are colored red, blue, magenta, and green, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Mouse monoclonal antibodies against myosin heavy chain (MyHC), CD38, and transforming growth factor-β (TGF-β), as well as rabbit polyclonal α-smooth muscle actin (α-SMA) antibodies, were purchased from R&D Systems (USA).

Techniques: Fluorescence, Marker, Expressing, Activation Assay, Immunostaining

Journal: bioRxiv

Article Title: Germ-free piglets display variable neuroinflammatory-like perturbations in prefrontal cortical microglia

doi: 10.64898/2026.03.22.713463

Figure Lengend Snippet: (A) Volcano plot of significant upregulated and downregulated genes in GF tissue, using p < 0.05 and a threshold of 1.2-fold change in expression as a cutoff for analysis. CD38 (p < 0.001, FC = 1.55), RSAD2 (p < 0.001, FC = 0.08), GABRR1 (p < 0.001, FC = 0.08), FN1 (p < 0.001, FC = 0.54), IFI6 (p < 0.001, FC = 0.37), IRAK1BP1 (p < 0.001, FC = 1.74), RGS1 (p < 0.001, FC = 3.22) are labeled in bold. (B) Graphical reconstruction of immune activation Ingenuity Pathway Analysis result, showing differentially expressed genes in our tissue related to this pathway. Panel made with BioRender. (C) Representative immunohistochemical images of the PFCII-III in control and GF animals. CD38 + (red), DAPI + (blue), and Iba1 + (white) staining (left, scale bar 200µm), a middle inset of the leftmost image (middle, scale bar 50µm), and a single channel (CD38) of this inset (right, scale bar 50µm) for each group n=3/animals per group. Data supported by Extended Data Table 6-1.

Article Snippet: Sections were blocked with 5% donkey serum, 1% BSA in PBST for 1 hour, and then stained with Iba1 (Wako, catalog #NC9288364), Ki67 (Abcam, catalog #ab16667), and CD38 (Proteintech, catalog #60006-1-Ig) overnight at 4°C.

Techniques: Expressing, Labeling, Activation Assay, Immunohistochemical staining, Control, Staining

Journal: Molecular Therapy Oncology

Article Title: A measles virus encoding a CD19/CD3 bispecific T cell engager shows enhanced preclinical anti-BCP-ALL efficacy without significant toxicity

doi: 10.1016/j.omton.2026.201127

Figure Lengend Snippet: MV-Blina infected cells secrete target-binding secBlina, which initiates ALL cell kill more efficiently than CD19/CD3-bsTE (A) Vero cells secrete secBlina. secBlina was harvested from the supernatant of infected Vero cells, followed by purification and concentration. Eluate (2 μg protein) was detected via His-tag by Western Blot in comparison to the commercial CD19/CD3-bsTE (1.8 μg). (B) Purified and concentrated secBlina binds specifically to target cells. Binding to target cells was confirmed by flow cytometry using CD3 − CD19 + REH, CD3 + CD19 - Jurkat, and CD3 − CD19 - K562 cell lines incubated with secBlina. Binding of secBlina to target cells was detected by flow cytometry via a His-tag antibody. (C) secBlina strongly binds to REH target cells. The REH cell line was first incubated with 1 μg secBlina. Increasing concentrations of CD19/CD3-bsTE were then added to displace cell-bound secBlina. Cell-bound secBlina was detected via an HA tag antibody by flow cytometry. (D) T cells in human PBMCs are activated by secBlina. Pooled PBMCs were stimulated with an (E) T cells are activated by secBlina during co-culture of PBMCs with REH cells. Pooled PBMCs were co-cultured with REH cells in a 1:1-ratio for 24 h while being treated with secBlina or the CD19/CD3-bsTE. Activated CD2 + CD69 + T cells were detected by flow cytometry. (F) MV-Blina infected leukemia cells produce and secrete secBlina. REH and Jurkat cells are shown 72 h post MV infection or Opti-MEM control. secBlina was detected by His-tag IF staining. Scale bars 50 μm. (G) secBlina effectively induces REH target cell kill compared to CD19/CD3-bsTE, whereas K562 control cells are unaffected. Target (T) cells REH (left panels) and negative control cells K562 (right panels) were incubated with secBlina (green) or CD19/CD3-bsTE (blue) for 24 h at indicated concentrations in the presence of PBMCs (effector; E) at different E:T ratios. Specific cell kill was determined by flow cytometry. Results in A, B, C, and F are shown as representatives of three independent experiments. Data are shown as mean ± SD of n = 3 (D, E) or n = 5 (G) independent experiments. Statistical analysis was performed using two-way ANOVA with Dunnett’s correction. ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; and ∗∗∗∗, p < 0.0001.

Article Snippet: For the displacement assay, 1 × 10 6 REH cells were incubated with 1 μg secBlina for 30 min, followed by increasing concentrations of a research-grade analog of the CD19/CD3-bsTE blinatumomab (BPS Bioscience) for an additional 30 min, all at 4°C. secBlina bound to target cells was detected using rabbit anti-HA antibody (clone EPR22819-101, 1:600, ab256483, Abcam) for 30 min at 4°C, followed by a goat-anti-rabbit IgG (H&L) AF488 (1:2000, ab150077, Abcam).

Techniques: Infection, Binding Assay, Purification, Concentration Assay, Western Blot, Comparison, Flow Cytometry, Incubation, Co-Culture Assay, Cell Culture, Control, Staining, Negative Control