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: Acta Neuropathologica

Article Title: Brachio-cervical inflammatory myopathy: multilevel clinical, histopathological and multi-omic analyses of a syndrome variably associated with systemic sclerosis

doi: 10.1007/s00401-026-03006-5

Figure Lengend Snippet: Immune pathology, B-cell characterization, and tertiary lymphoid organ-related features. a Diffuse relatively mild sarcolemmal MHC cl. 1 positivity and presence of many MHC cl. 1 positive lymphomonocytic cells in the endomysium, and in a B-cell cluster (arrow). MHC cl. 1 immunohistochemistry; original magnification ×200. b Mild and infrequent sarcolemmal MHC cl. 2 positivity and presence of many MHC cl. 2 positive lymphomonocytic cells in the endomysium, and in a B-cell cluster (arrow). MHC cl. 2 immunohistochemistry; original magnification ×200. c Sarcolemmal complement deposits (arrow) and presence of single necrotic myofibers (arrowhead) that show sarcoplasmic complement deposits. C5b-9 (MAC) immunohistochemistry; original magnification ×200. d Presence of numerous endomysial macrophages and occasional myophagocytoses. CD68 immunohistochemistry; original magnification ×200. e Scattered, individual endomysial T cells. CD8 immunohistochemistry; original magnification ×200. f Focally clustering B cells with a roundish appearance reminiscent of so-called tertiary lymphoid organs (TLOs). CD20 immunohistochemistry; original magnification ×200. g , h TLO-like immune cell clusters show compartmentalization with bcl2-positive B-cell nuclei at the margins and bcl6-positive nuclei in their (germinal) center. Bcl2 (G) and bcl6 (H) immunohistochemistry; original magnification ×600. i Numerous Ki67 + proliferating cells are focally clustering predominantly in germinal centers of TLOs. Ki67 immunohistochemistry; original magnification ×600. j A B-cell cluster contains some CD138-positive plasma cells. CD138 immunohistochemistry; original magnification ×200. k Similarly, those plasma cells and a few B cells are identified by nuclear MUM1 positivity in the light zone of a germinal center. MUM1 immunohistochemistry; original magnification ×200. l CD38-positive plasmablasts are partly proliferating and show co-immunoreactivity with Ki67. CD38 AF488 (green) and Ki67 Cy3 (red) immunofluorescence; original magnification ×200. a – l Scale bar represents 100 µm

Article Snippet: The antibodies used for immunohistochemistry included C5b-9 (Dako, aE11, 1:200), CD8 (Dako, C8/144B, 1:100), CD20 (Dako, Ks 20.8 1:200), Ki-67 (Dako, Mib-1, 1:100), MxA (Millipore, MABF938, 1:100), CD38 (Novocastra SPC32, 1:100), MUM1 (Dako, MUM1P, 1:50), CD45 (Dako, 2B11, 1:400), CD56 (Serotec/MCA591 ERIC-1, 1:400), CD68 (Dako, EBM11, 1:100), MHC class I (Dako, w6/32, 1:1000), MHC class II (Dako, CR3/43, 1:100), and SQSTM1/p62 (Abcam, polyclonal, 1:100).

Techniques: Cell Characterization, Immunohistochemistry, Clinical Proteomics, Immunofluorescence