Skip to main content
. 2006 Jul 18;3(8):e253. doi: 10.1371/journal.pmed.0030253

Figure 7. Lipoproteins of Patient Serum Are Endocytosed by Neutrophils and Lead to Intracellular Accumulation of Neutral Lipids and Enhanced Neutrophil Apoptosis.

Figure 7

(A, B, and C) Freshly isolated human PMNs were incubated for 16 h with control (C) and patient (P, P1, P2, and Pat) sera. For (A and B), cells were washed extensively and cell lysates probed for ApoB and LDL-receptor by SDS-PAGE and Western blotting. A proteolytic fragment [56] of ApoB was detected at ~80 kDa. LDL receptor served as a load control. For (B and C), the FcγRIIa blocking mab IV.3 was added during the incubation (P1+ [B]; Pat. + mab IV.3 [C]). For (C), cells were fixed, stained with Nile Red and subjected to confocal fluorescence microscopy.

(D) PMNs of control or schistosome-infected hamster were stained with Nile Red and subjected to confocal fluorescence microscopy. Data are representative of four infected and control animals.

(E) PMNs were incubated for 20 h with 20% (left) or 10% (right) control or patient serum as indicated. For the right bar graph, 10 μg/ml normal rabbit antibody (Control) or anti-ApoB antibody were preincubated with control serum, and subsequently added to PMNs. Cells were counted before and after incubation with serum, stained with Annexin V-FITC (which indicates apoptosis), and analysed by FACS. Results are expressed as percentage of Annexin V-negative cells, relative to the number of cells before incubation with serum. Statistical analysis was done by Student's t-test, ** p < 0.01.