Journal: Frontiers in Immunology

Article Title: Molecular determinants of STEC-HUS: from complement activation to microvascular thrombosis

doi: 10.3389/fimmu.2026.1749811

Figure Lengend Snippet: Prothrombotic effects of STEC-HUS serum are dependent on WPB exocytosis from endothelial cells. P-selectin (A) and vWF (B) expression on unstimulated HMEC-1 exposed to serum from patients with acute STEC-HUS (n = 5 for P-selectin experiments; n = 6 for vWF experiments), in the presence or in the absence of different complement inhibitors (sCR1, 150 µg/mL; factor B inhibitor, iptacopan,10 µM; eculizumab 100 µg/mL) or to a pool of control sera (normal human serum, NHS), run in parallel. Results are shown as pixel 2 /high-power field (HPF) of stained surface area. Data are mean ± SD. Circles represent single patients’ data. The addition of either sCR1, or iptacopan or eculizumab to patient’s serum significantly decrease both P-selectin and vWF expression induced by patient’s serum alone. *P < 0.05 vs STEC-HUS serum alone (ANOVA, followed by Tukey’s multiple comparisons test for data of P-selectin expression and by Holm-Šídák’s multiple comparisons test for data of vWF expression). (C) Endothelial surface area covered by thrombi on ADP-activated HMEC-1 exposed to serum from STEC-HUS patients collected during the acute phase of the disease and then perfused with whole blood. Before the experiments, HMEC-1 were left for 16 hours with medium added or not with the RalA inhibitor BQU57 (10 µM). Data are expressed as mean ± SD of percentages of serum-induced thrombus formation in respect to a pool of control sera (normal human serum, NHS), run in parallel in each experiment and set as 100% (n = 3 independent experiments). Circles indicate single patients’ data. Horizontal dashed lines indicate upper and lower limits of the normal range . *P < 0.05 (paired Student’s t test). (D) Representative confocal microscopy images (original magnification X200) of experiments of thrombus formation (green staining) relative to Figure 6C . Scale bar: 100 µm.

Article Snippet: Cells were washed again and treated with the following specific antibodies: FITC-conjugated rabbit anti-human C3c-complement (Dako, that recognizes C3c, part of C3 and C3b, 1:300 final dilution in Dapi 1 μg/mL); or rabbit anti-human complement C5b-9 complex (Calbiochem, 1:200 final dilution in PBS1X) followed by FITC-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:50 final dilution in 1 μg/mL Dapi); or goat anti-human C4 (Abcam, 1:100 final dilution in PBS1X) followed by Cy3-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:200 final dilution in Dapi 1 μg/mL); or FITC-conjugated anti-human IgG (Sigma Aldrich, 1:32 final dilution in 1 μg/mL Dapi); or mouse anti-human P-selectin (R&D System, 20 μg/mL final concentration in PBS1X), followed by Cy3-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:60 final dilution in 1 μg/mL Dapi); or rabbit anti-human vWF (Dako, 10 μg/mL final concentration in PBS1X), followed by Cy3-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:50 final dilution in 1 μg/mL Dapi).

Techniques: Expressing, Control, Staining, Confocal Microscopy

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Mapping the expression of endothelial adhesion receptors for Plasmodium falciparum -infected erythrocytes in fatal cerebral malaria in Malawian children

doi: 10.1093/jnen/nlaf104

Figure Lengend Snippet: Plots demonstrate the average sequestration ratio in the CM2 cases. The ratio is higher for CD36, VCAM-1, E-selectin, and TSP as compared with ICAM-1. Abbreviations: CM, cerebral malaria; TSP, thrombospondin.

Article Snippet: The expression of the endothelial adhesion receptors ICAM-1, VCAM-1, E-selectin, CD36, and TSP in frozen sections and smears was detected by immunohistochemistry with the avidin-biotin technique using monoclonal antibodies specific for ICAM-1 (CD54, 1:100; mouse monoclonal; Clone 15.2; Bio-Rad AbD Serotec), VCAM-1 (CD106, 1:35; mouse monoclonal; Clone 1G11; Immunotech), E-selectin (CD62E, 1:75; mouse monoclonal; Clone BB1G-E4; R&D Systems), CD36 (1:30; mouse monoclonal; Clone FA6-152; Immunotech), and TSP (CD47, 1:50; mouse monoclonal; Clone P10; Immunotech).

Techniques:

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Mapping the expression of endothelial adhesion receptors for Plasmodium falciparum -infected erythrocytes in fatal cerebral malaria in Malawian children

doi: 10.1093/jnen/nlaf104

Figure Lengend Snippet: (A-E) Expression of the 5 cytoadhesion receptors by microvascular endothelial cells in CM2 patients. Receptor expression was variable along the length of a microvessel and among microvessels. (A) ICAM-1 is the most widely and highly expressed receptor. Occasional endothelial cells show no staining (arrow). (B) VCAM-1 is focally expressed by endothelial cells of microvessels containing iRBC. Adjacent endothelial cells show no staining. (C) A few E-selectin-positive endothelial cells along the length of this microvessel. (D) Scattered CD36-expressing endothelial cells alternate with cells not expressing this receptor. (E) Infrequent expression of TSP by microvascular endothelium (arrows). Abbreviations: CM, cerebral malaria; iRBC, infected red blood cells.

Article Snippet: The expression of the endothelial adhesion receptors ICAM-1, VCAM-1, E-selectin, CD36, and TSP in frozen sections and smears was detected by immunohistochemistry with the avidin-biotin technique using monoclonal antibodies specific for ICAM-1 (CD54, 1:100; mouse monoclonal; Clone 15.2; Bio-Rad AbD Serotec), VCAM-1 (CD106, 1:35; mouse monoclonal; Clone 1G11; Immunotech), E-selectin (CD62E, 1:75; mouse monoclonal; Clone BB1G-E4; R&D Systems), CD36 (1:30; mouse monoclonal; Clone FA6-152; Immunotech), and TSP (CD47, 1:50; mouse monoclonal; Clone P10; Immunotech).

Techniques: Expressing, Staining, Infection

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Mapping the expression of endothelial adhesion receptors for Plasmodium falciparum -infected erythrocytes in fatal cerebral malaria in Malawian children

doi: 10.1093/jnen/nlaf104

Figure Lengend Snippet: (A-E) Expression of the 5 cytoadhesion receptors by microvascular endothelial cells in CM1 patients. (A) ICAM-1 is expressed by a greater number of endothelial cells as compared with VCAM-1 (B), E-selectin (C), CD36 (D), and TSP (E). Similar to the CM2 cases, not all endothelial cells of a microvessel and not all microvessels containing iRBC express cytoadhesion receptors. Abbreviations: CM, cerebral malaria; iRBC, infected red blood cells.

Article Snippet: The expression of the endothelial adhesion receptors ICAM-1, VCAM-1, E-selectin, CD36, and TSP in frozen sections and smears was detected by immunohistochemistry with the avidin-biotin technique using monoclonal antibodies specific for ICAM-1 (CD54, 1:100; mouse monoclonal; Clone 15.2; Bio-Rad AbD Serotec), VCAM-1 (CD106, 1:35; mouse monoclonal; Clone 1G11; Immunotech), E-selectin (CD62E, 1:75; mouse monoclonal; Clone BB1G-E4; R&D Systems), CD36 (1:30; mouse monoclonal; Clone FA6-152; Immunotech), and TSP (CD47, 1:50; mouse monoclonal; Clone P10; Immunotech).

Techniques: Expressing, Infection

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Mapping the expression of endothelial adhesion receptors for Plasmodium falciparum -infected erythrocytes in fatal cerebral malaria in Malawian children

doi: 10.1093/jnen/nlaf104

Figure Lengend Snippet: (A) Plots demonstrating the average number of receptor-expressing endothelial cells per 1000-µm microvessel length in the CM1 cases. The expression of ICAM-1 is greater compared with that of VCAM-1, E-selectin, CD36, and TSP. (B) Plots illustrating the average number of receptor-expressing endothelial cells per 1000 µm microvessel length in the CM2 cases. As compared with ICAM-1, the expression of VCAM-1, E-selectin, and TSP is lower, whereas that of CD36 is slightly higher in both cortical and subcortical areas. Abbreviations: CM, cerebral malaria; TSP, thrombospondin.

Article Snippet: The expression of the endothelial adhesion receptors ICAM-1, VCAM-1, E-selectin, CD36, and TSP in frozen sections and smears was detected by immunohistochemistry with the avidin-biotin technique using monoclonal antibodies specific for ICAM-1 (CD54, 1:100; mouse monoclonal; Clone 15.2; Bio-Rad AbD Serotec), VCAM-1 (CD106, 1:35; mouse monoclonal; Clone 1G11; Immunotech), E-selectin (CD62E, 1:75; mouse monoclonal; Clone BB1G-E4; R&D Systems), CD36 (1:30; mouse monoclonal; Clone FA6-152; Immunotech), and TSP (CD47, 1:50; mouse monoclonal; Clone P10; Immunotech).

Techniques: Expressing

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Mapping the expression of endothelial adhesion receptors for Plasmodium falciparum -infected erythrocytes in fatal cerebral malaria in Malawian children

doi: 10.1093/jnen/nlaf104

Figure Lengend Snippet: Plots showing the average number of colocalized microvessels per 10 microvessels in the CM2 cases. As compared with ICAM-1, the colocalization ratio is higher for CD36 and lower for VCAM-1, E-selectin and TSP. Abbreviations: CM, cerebral malaria; TSP, thrombospondin.

Article Snippet: The expression of the endothelial adhesion receptors ICAM-1, VCAM-1, E-selectin, CD36, and TSP in frozen sections and smears was detected by immunohistochemistry with the avidin-biotin technique using monoclonal antibodies specific for ICAM-1 (CD54, 1:100; mouse monoclonal; Clone 15.2; Bio-Rad AbD Serotec), VCAM-1 (CD106, 1:35; mouse monoclonal; Clone 1G11; Immunotech), E-selectin (CD62E, 1:75; mouse monoclonal; Clone BB1G-E4; R&D Systems), CD36 (1:30; mouse monoclonal; Clone FA6-152; Immunotech), and TSP (CD47, 1:50; mouse monoclonal; Clone P10; Immunotech).

Techniques:

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Mapping the expression of endothelial adhesion receptors for Plasmodium falciparum -infected erythrocytes in fatal cerebral malaria in Malawian children

doi: 10.1093/jnen/nlaf104

Figure Lengend Snippet: Plots demonstrate the average number of colocalized microvessels per 10 microvessels in the CM1 cases. The colocalization ratio for ICAM-1 is higher as compared with that of VCAM-1, E-selectin, and TSP and not very different from that of CD36. Abbreviations: CM, cerebral malaria; TSP, thrombospondin.

Article Snippet: The expression of the endothelial adhesion receptors ICAM-1, VCAM-1, E-selectin, CD36, and TSP in frozen sections and smears was detected by immunohistochemistry with the avidin-biotin technique using monoclonal antibodies specific for ICAM-1 (CD54, 1:100; mouse monoclonal; Clone 15.2; Bio-Rad AbD Serotec), VCAM-1 (CD106, 1:35; mouse monoclonal; Clone 1G11; Immunotech), E-selectin (CD62E, 1:75; mouse monoclonal; Clone BB1G-E4; R&D Systems), CD36 (1:30; mouse monoclonal; Clone FA6-152; Immunotech), and TSP (CD47, 1:50; mouse monoclonal; Clone P10; Immunotech).

Techniques: