Properties of the new Kenyan rosetting lines.
<p>A) IgM binding by KE10R+ infected erythrocytes detected by flow cytometry. Forward and side scatter were used to gate on erythrocytes and exclude debris (left panel) and PfEMP1-expressing infected erythrocytes were detected by staining with 20μg/ml of polyclonal rabbit IgG against NTS-DBLα...
محفوظ في:
| المؤلف الرئيسي: | |
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| مؤلفون آخرون: | , , , , , , , , |
| منشور في: |
2025
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| الموضوعات: | |
| الوسوم: |
إضافة وسم
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| الملخص: | <p>A) IgM binding by KE10R+ infected erythrocytes detected by flow cytometry. Forward and side scatter were used to gate on erythrocytes and exclude debris (left panel) and PfEMP1-expressing infected erythrocytes were detected by staining with 20μg/ml of polyclonal rabbit IgG against NTS-DBLα of KE10VAR_R1 followed by 1/1000 dilution of Alexa Fluor 647-conjugated goat anti-rabbit IgG secondary antibody and 1/2500 dilution of Vybrant DyeCycle Violet (middle panel Q2). IgM staining of the Q2 cell population was detected with 1/1000 dilution of an Alexa Fluor 488-conjugated goat anti-human IgM heavy chain antibody (red). The negative control (blue) was parasites grown in IgM-depleted medium and stained with the same antibodies. Results are representative of two independent experiments. B) Effect of low-dose trypsinisation on rosetting. Purified infected erythrocytes were treated with 0.5, 1 or 5 μg/ml of trypsin for 5 mins and the rosette frequency relative to a control with no added enzyme was calculated. The mean and standard deviation from three independent experiments per parasite line is shown. The rosette frequency of the untreated control was between 46%-81% (KE10R+), 45%-89% (PC0053R+), 16%-30% (KE11R+) and 61%-91% (KE08R+). C-E) Effect of heparin on rosetting. The mean and standard error from two or three independent experiments per parasite line is shown.</p> |
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