Journal: mBio

Article Title: Plasmodium vivax Infection Alters Mitochondrial Metabolism in Human Monocytes

doi: 10.1128/mBio.01247-21

Figure Lengend Snippet: Glucose uptake was increased in monocyte subsets during P. vivax infection. (A) Glucose uptake measured ex vivo in CD14 + CD16 – (green), CD14 + CD16 + (red), and CD14 low CD16 + (blue) monocytes from HD (open symbols) and Pv (solid symbols). (B) Level of glucose uptake by monocyte subsets from malaria patients. (C) Extracellular flux analysis using OCR to measure bioenergetics parameters associated with mitochondrial function. Injections of oligomycin, FCCP, and rotenone plus antimycin A were used to perturb mitochondrial functions in circulating monocytes from HD and Pv stimulated with PMA. Radar graph of HD (open orange circles) and Pv (solid orange circles) shows mitochondrial stress parameters normalized to baseline for nonstimulated cells. Cumulative data of at least four individual experiments. Scatterplot with bars representing the mean ± standard deviation (SD) and histogram representing median fluorescence intensity (MFI) of NBDG on the x axis; each curve represents a monocyte subset of a single donor normalized to mode 100% on the y axis. P values were calculated by two-tail Mann-Whitney test (A, HD n = 4, Pv n = 10; C, HD n = 7, Pv n = 11). *, P ≤ 0.05; **, P ≤ 0.01.

Article Snippet: Total monocytes were purified by positive selection by means of magnetic separation using anti-CD14 microbeads according to the manufacturer’s instructions (MACS; Miltenyi Biotec), or CD14 + CD16 − , CD14 + CD16 + , and CD14 low CD16 + monocyte subsets were purified with a FACSAria II cell sorter (BD Biosciences), using the following antibodies in different combinations: anti-CD14 (clone 61D3)-APC, anti-CD14 (clone 61D3)-PE (eBioscience), anti-CD16 (clone 3G8)-PerCPCy5.5, anti-CD16 (clone 3G8)-PECy7, and anti-CD66b (clone G10F5)-FITC (BD Bioscience).

Techniques: Infection, Ex Vivo, Standard Deviation, Fluorescence, MANN-WHITNEY

Journal: mBio

Article Title: Plasmodium vivax Infection Alters Mitochondrial Metabolism in Human Monocytes

doi: 10.1128/mBio.01247-21

Figure Lengend Snippet: Plasmodium vivax infection alters mitochondrial contents in monocyte subsets. (A) Bar graphs showing ex vivo mitochondrial content and mass based on MFI of MitoTracker green (bottom panel) and MitoTracker red CMX-Ros (top panel) reactivity, respectively, in CD14 + CD16 − (green), CD14 + CD16 + (red), and CD14 low CD16 + (blue) monocytes from HD (open symbols) and Pv (solid symbols). (B) Bar graphs (left panels) and representative histograms (right panel) showing MFI of MitoTracker green (top panel) and MitoTracker red CMX-Ros (bottom panel) reactivity in monocyte subsets from malaria patients. Cumulative data of at least four individual experiments. Scatterplot with bars representing the mean ± SD and histogram representing the MFI of MitoTracker green and MitoTracker red on the x axis; each curve represents a monocyte subset of a single donor normalized to mode 100% on the y axis. P values were calculated by (A) an unpaired t test (HD n = 5 and n = 4, Pv n = 6); (B) one-way ANOVA with Bonferroni’s multiple-comparison test ( n = 18). *, P ≤ 0.05; ***, P ≤ 0.001.

Article Snippet: Total monocytes were purified by positive selection by means of magnetic separation using anti-CD14 microbeads according to the manufacturer’s instructions (MACS; Miltenyi Biotec), or CD14 + CD16 − , CD14 + CD16 + , and CD14 low CD16 + monocyte subsets were purified with a FACSAria II cell sorter (BD Biosciences), using the following antibodies in different combinations: anti-CD14 (clone 61D3)-APC, anti-CD14 (clone 61D3)-PE (eBioscience), anti-CD16 (clone 3G8)-PerCPCy5.5, anti-CD16 (clone 3G8)-PECy7, and anti-CD66b (clone G10F5)-FITC (BD Bioscience).

Techniques: Infection, Ex Vivo

Journal: mBio

Article Title: Plasmodium vivax Infection Alters Mitochondrial Metabolism in Human Monocytes

doi: 10.1128/mBio.01247-21

Figure Lengend Snippet: Altered mitochondrial function in monocytes from P. vivax -infected patients is accompanied by induction of PKM, mitoferrin-1, and SOD2. (A) mRNA counts of genes involved in glycolysis (PKM), OXPHOS (complexes II, IV, and V), and complex IV insertion in inner mitochondrial membrane in CD14 + CD16 – (green bar), CD14 + CD16 + (red bar), and CD14 low CD16 + (blue bar) monocytes from Pv and HD. (B) mRNA counts of genes involved in TIM complex and TOM complex assembly, mitoferrin-1 (SLC25A37), and SOD2 in monocyte subsets from Pv and HD. Classical monocytes are represented in green, inflammatory monocytes in red, and patrolling monocytes in blue. Scatterplot with bars representing the mean ± SD. P values were calculated by two-tailed Mann-Whitney test. (A) PKM, complexes II, IV, and V (HD n = 7 or 8, Pv n = 8), COX 10 and 18 (HD n = 3, Pv n = 5); (B) (HD n = 3, Pv n = 5). *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.

Article Snippet: Total monocytes were purified by positive selection by means of magnetic separation using anti-CD14 microbeads according to the manufacturer’s instructions (MACS; Miltenyi Biotec), or CD14 + CD16 − , CD14 + CD16 + , and CD14 low CD16 + monocyte subsets were purified with a FACSAria II cell sorter (BD Biosciences), using the following antibodies in different combinations: anti-CD14 (clone 61D3)-APC, anti-CD14 (clone 61D3)-PE (eBioscience), anti-CD16 (clone 3G8)-PerCPCy5.5, anti-CD16 (clone 3G8)-PECy7, and anti-CD66b (clone G10F5)-FITC (BD Bioscience).

Techniques: Infection, Two Tailed Test, MANN-WHITNEY