Journal: Frontiers in Immunology

Article Title: The power of GM-CSF: immune regulation in the defense against Phialophora verrucosa infection

doi: 10.3389/fimmu.2025.1662183

Figure Lengend Snippet: Immune cell infiltration and cytokine levels in WT and Csf2 KO mice during P. verrucosa infection. Immunohistochemical staining of macrophage marker CD68 (A) and neutrophil marker myeloperoxidase (MPO) (B) in footpad tissues from the injection site of mice infected with P. verrucosa for up to 2 weeks. * P < 0.05. WT mice and Csf2 KO mice were subcutaneously inoculated with 5×10 7 live conidia in both hind footpads. Cytokine levels in footpad tissue homogenates (n=3) (C) and serum (n=4) (D) were measured by flow cytometric bead array at weeks 0, 1, 2, 3, and 4 post-infection. Data are representative of two independent experiments. * P < 0.05, ** P < 0.01; n.s. indicates no significant difference.

Article Snippet: For immunohistochemical analysis, tissue sections were incubated with primary antibodies against the macrophage marker CD68 (Servicebio) and neutrophil marker myeloperoxidase (MPO, Abcam).

Techniques: Infection, Immunohistochemical staining, Staining, Marker, Injection

Journal: Inflammation Research

Article Title: Quantitative temporal analysis of pancreatic islet T lymphocyte and macrophage infiltration heralded by serum IgE in congenic BioBreeding (BB) Gimap5 − / − rats at risk for insulitis and acute onset diabetes

doi: 10.1007/s00011-025-02101-9

Figure Lengend Snippet: Spatial infiltration of CD3 + and ED1 + immune cells in pancreatic islets. A Representative image of CD3+ T cell infiltration in a sBBM Gimap5-DP rat. The perimeter of the analyzed islet is marked in green, defined using a convolutional neural network (CNN)-based segmentation algorithm. B Schematic illustrating the spatial analysis of infiltrating immune cells. The green line indicates the islet boundary. Analysis was performed in 10 μm intervals extending up to 100 μm into the islets interior (negative values) and 100 μm into the surrounding exocrine tissue (positive values), reltive to the islet perimeter (0 μm). C Quantification of infiltrating CD3+ T cells (top row) and ED1+ macrophages (bottom row) shown as cell density (cells/mm 2 ) in 10 μm segments. Data are presented for sBBM Gimap5 + / + (Gimap5-DR) rats (n = 5; red bars), Gimap5 − / − (Gimap5-DP) prior to diabetes onset (n = 8; green bars), and Gimap5 − / − (Gimap5-DP) rats at the time of clinical onset of diabetes (n = 4; blue bars). Data are presented as mean ± SD

Article Snippet: Sections at depths of 15, 40, 65, 225, 250, 275, 435, 460, and 485 μm were stained for CD3, a marker of T lymphocytes (CD3 antibody (clone SP7), diluted 1:500, product no ab 16,669, Abcam, Cambridge, UK) and at 20, 45, 70, 230, 255, 280, 440, 465, and 490 μm were stained for ED1, a macrophage marker (mouse anti-rat CD68, diluted 1:100, MCA341R, Bio-Rad, Hercules, CA, USA).

Techniques:

Journal: The FASEB Journal

Article Title: ATF6 Alleviates Endothelial Inflammation Following Extended Hepatectomy Through Inhibition of TRIM10 / NF ‐ κB Signaling

doi: 10.1096/fj.202402197RRR

Figure Lengend Snippet: Expression of unfolded protein response (UPR) genes following extended hepatectomy (PH80) and major hepatectomy (PH70). (A) Transcriptomic analysis of the expression of UPR genes (atf6, atf4, xbp1, and hspa5/GRP78) at 0 h (control), 1 h, 6 h, 12 h, 1 day, and 3 days after PH80 and PH70 in rats (* p < 0.05 between PH80 and PH70, N = 3 at each time point); (B) the expression of ATF6 and cleaved ATF6 (cATF6) following PH80 and PH70 in mice determined by western blot analysis and semiquantitative analysis ( N = 5). (C) Western blot analysis of the expression of XBP1, spliced XBP1 (XBP1s) and ATF4 following PH80 and PH70 in mice and semiquantitative analysis ( N = 5); (D) western blot analysis of GRP78 and GRP94 expression following PH80 and PH70 treatment in mice and semiquantitative analysis of GRP78 and GRP94 expression ( N = 5); (E) immunohistochemical analysis of hepatic ATF6 expression following PH80 and PH70 treatment in mice (original magnification ×400, scale bars: 50 μm) and semiquantitative analysis of the results ( N = 5); (F) assessment of hepatic ATF6 expression following marginal hepatectomy ( N = 8) and minor hepatectomy ( N = 6) in humans by immunohistochemistry (original magnification ×400, scale bars: 50 μm) and semiquantitative analysis; (G) hepatic ATF6 expression (green) in liver sinusoidal endothelial cells (CD31, purple), macrophages (CD68, orange) and Th cells (CD4, white) in the early stage after extended hepatectomy in mice by immunofluorescence (original magnification ×400, scale bars: 20 μm).

Article Snippet: Immunostaining was conducted with primary antibodies against ATF6 (1:100, ab122897, Abcam, Cambridge, UK), Ki‐67 (1:200, ab15580, Abcam, Cambridge, UK), LSEC marker CD31 (1:100, 11265‐1‐AP, Proteintech, Wuhan, China), macrophage marker CD68 (1:100, 25747‐1‐AP, Proteintech, Wuhan, China), T helper cell marker CD4 (1:100, 65104‐1‐AP, Proteintech, Wuhan, China), and TRIM10 (1:200, bs‐9409R, Bioss, Beijing, China).

Techniques: Expressing, Control, Western Blot, Immunohistochemical staining, Immunohistochemistry, Immunofluorescence

Journal: Gut

Article Title: DYRK1B blockade promotes tumoricidal macrophage activity in pancreatic cancer

doi: 10.1136/gutjnl-2023-331854

Figure Lengend Snippet: Intratumoral macrophage abundance correlates with DYRK1B levels in human PDAC. (A) Immunohistochemical CD68 staining (brown) of human PDAC tissue microarrays (Marburg cohort). DYRK1B levels were determined by bulk RNAseq. (B) Quantification of CD68 immunohistochemistry intensity in patients with PDAC, which were split into DYRK1B -low and high subgroups (n=15 each). Each dot represents one patient tumour (mean±SD). (C) CD24 mRNA levels in patients with PDAC as assessed by bulk RNA sequencing. Patients were split into DYRK1B -low/high subgroups (n=15 each). Each dot represents one patient tumour (mean±SD). (D) Correlation between CD68 and DYRK1B bulk mRNA expression in patients with PDAC of the TCGA cohort. (E) Correlation between MSR1 and DYRK1B bulk mRNA expression in patients with PDAC of the TCGA cohort. (F) Correlation between ITGAM (encoding CD11B) and DYRK1B bulk mRNA expression in patients with PDAC of the TCGA cohort. DYRK1B, dual specificity and tyrosine phosphorylation-regulated kinase 1B; PDAC, pancreatic ductal adenocarcinoma; mRNA, messenger RNA; TCGA, The Cancer Genome Atlas.

Article Snippet: To this end, we first made use of the Marburg cohort of patients with PDAC, whose tissue was stained by immunohistochemistry for the pan-macrophage marker CD68 (n=46).

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, RNA Sequencing, Expressing, Phospho-proteomics

Journal: BMC Veterinary Research

Article Title: Characterization of giant endocrine cells in the fundic stomach of African catfish (Clarias gariepinus) demonstrated by histochemical, immunohistochemical and ultrastructure microscopy methods suggesting their role in immunity

doi: 10.1186/s12917-024-04237-y

Figure Lengend Snippet: Identity, sources, and working dilution of antibodies used in the present immunohistochemical analysis

Article Snippet: CD68 (Macrophage Marker) Ab-3 (Clone KP1) , Mouse Anti-CD68 (Thermo Fisher Scientific Lab Vision Corporation, Fremont, USA) , Mouse Monoclonal Antibody Cat. #MS-397-R7 , 1:100 , Over night , boiling in citrate buffer (pH 6.0), 20 min , Goat anti-rabbit secondary antibody (Cat. no. K4003, EN Vision + TM System Horseradish Peroxidase Labelled Polymer; Dako). Ready to use (30 min at room temperature).

Techniques: Immunohistochemical staining, Incubation, Marker, Polymer

Journal: BMC Veterinary Research

Article Title: Characterization of giant endocrine cells in the fundic stomach of African catfish (Clarias gariepinus) demonstrated by histochemical, immunohistochemical and ultrastructure microscopy methods suggesting their role in immunity

doi: 10.1186/s12917-024-04237-y

Figure Lengend Snippet: Shows the immune reactivity of the endocrine cells with CD68. The fundic stomach paraffin section was immune stained with CD68. A : Endocrine cells positive for CD68 (shown by arrows) are seen in the gastric gland (gl), lamina propria (lp), and submucosa (sb). B : Endocrine cells positive for CD68 (shown by arrows) formed clusters within the sub-epithelial lymphatic space (ls). C , D : CD68-positive endocrine cells (shown by arrows) are situated in the submucosa (sb)

Article Snippet: CD68 (Macrophage Marker) Ab-3 (Clone KP1) , Mouse Anti-CD68 (Thermo Fisher Scientific Lab Vision Corporation, Fremont, USA) , Mouse Monoclonal Antibody Cat. #MS-397-R7 , 1:100 , Over night , boiling in citrate buffer (pH 6.0), 20 min , Goat anti-rabbit secondary antibody (Cat. no. K4003, EN Vision + TM System Horseradish Peroxidase Labelled Polymer; Dako). Ready to use (30 min at room temperature).

Techniques: Paraffin Section, Staining

Journal: Clinical and Translational Medicine

Article Title: Cyclic‐di‐GMP induces inflammation and acute lung injury through direct binding to MD2

doi: 10.1002/ctm2.1744

Figure Lengend Snippet: MD2 deficiency prevents CDG‐induced lung injury in vivo. (A) Lung wet/dry weight ratio ( n = 6 in each group, biological replicates). (B) Quantification of the lung injury scores ( n = 6 in each group, biological replicates). (C) Total cell counts in BALF samples were measured using a hemocytometer ( n = 6 in each group, biological replicates). (D) Total protein concentration in BALF samples was measured ( n = 6 in each group, biological replicates). (E) MPO activity levels in lung lysates ( n = 6 in each group, biological replicates). (F) Neutrophils in BALF samples were assessed using Wright‐Giemsa staining ( n = 6 in each group, biological replicates). (G) Representative H&E‐staining of lung tissues. Scale bar: 50 µm. (H) Immunohistochemical staining of lung tissues for CD68 macrophage markers. Scale bar: 50 µm. (I) mRNA levels of adhesion factors Icam1 and Vcam1 in lung tissues were measured via RT‐qPCR assay. Data were normalized to the levels of Actb ( n = 6 in each group, biological replicates). (J) The protein levels of ICAM1 and VCAM1 were examined in lung tissues. GAPDH was used as the loading control. Data information: Data are presented as mean ± SEM. One‐way ANOVA followed by Dunnett's multiple comparisons test.

Article Snippet: Antibodies against TLR4 (sc‐293072), MD2 (sc‐80183), and macrophage markers CD68 (sc‐20060) were purchased from Santa Cruz Biotechnology.

Techniques: In Vivo, Protein Concentration, Activity Assay, Staining, Immunohistochemical staining, Quantitative RT-PCR, Control