Plasma fatty acid distribution (%) at day 10 post infection.

<p>^∞ PI: Peroxidability index of lipids. The value is calculated based on the relative oxidation rate of unsatured fatty acids as follows:</p><p>PI = (%monoenoic x 0.025)+(%dienoic x 1)+(%trienoic x 3)+(%tetraenoic x 4)+ +(%pentaenoic x 6)+(hexaenoic x 8)</p><p>*p<0.05;</p><p>**p<0.01;</p><p>*** p<0.0001 vs CTR</p><p>n = 5</p><p>Plasma fatty acid distribution (%) at day 10 post infection.</p

Alteration in the lipid profiles of the lungs from uninfected, <i>Pb</i>NK65 or <i>Pc</i>AS infected mice.

<p>Mice infected with <i>Pb</i>NK65 (treated or not intraperitoneally with DEX 80mg/kg) or <i>Pc</i>AS were perfused and dissected at day 8 and 10 post infection. The total content of PL (panel A), ChoE (panel B), PC (panel C) /mg protein were determined as described in Materials and Methods. n = 6 mice for each condition; *p<0.05; **p<0.01; ***p<0.0001 vs CTR; °p< 0.05; °°p<0.01; °°°p<0.0001 vs <i>Pc</i>AS for each time point; $ p<0.05 <i>Pb</i>NK65-DEX vs <i>Pb</i>NK65 day 10. PL = phospholipids; ChoE = Cholesterol Esters; PC = phosphatidylcholine.</p

MOESM1 of Differential induction of malaria liver pathology in mice infected with Plasmodium chabaudi AS or Plasmodium berghei NK65

Additional file 1. ALT and AST determination in mice infected with P. berghei NK65 or P. chabaudi AS. C57BL/6J mice were injected intraperitoneally with 104 erythrocytes infected with P. berghei NK65 or P. chabaudi AS. Serum levels of AST (panel A), ALT (panel B) and the AST/ALT ratio (panel C) were determined at day 8 and 10 post infection according to manufacturer’s protocol (Teco Diagnostics, California, USA). n = 3-6 mice for each time point and strain, additional data can be found in [16]. *p < 0.05; **p < 0.01 versus control

Alterations in the lipid profile of the pulmonary surfactant of <i>Pb</i>NK65 or <i>Pc</i>AS infected mice.

<p>(Panel A) Large aggregate (LA) and (Panel B) small aggregate (SA) fractions of pulmonary surfactant were isolated from the BAL fluid of control mice and mice infected with <i>Pb</i>NK65 or <i>Pc</i>AS and the percentage distribution of PL was determined. SM = Sphingomyelin, PE = phosphatidylethanolamine, PG = phosphatidylglycerol and LPC = lysophosphatidylcholine. n = 5–7; * p< 0.05 vs CTR; ** p<0.001 vs CTR; ° p< 0.05, °° p< 0.01 vs <i>Pc</i>AS for each time point.</p

Protein content of the Large Aggregate (LA) fraction of BAL fluid from uninfected, <i>Pb</i>NK65 or <i>Pc</i>AS infected mice.

<p>Bronchoalveolar lavage (BAL) fluid samples were collected from control mice and mice infected with <i>Pb</i>NK65 (n = 10 for day 8 post infection, n = 13 for day 10 post infection), or <i>Pc</i>AS (n = 8 for day 8 or 10 post infection). The large aggregate fraction (LA) was isolated and the content of proteins was determined. *p<0.05; **p<0.01; ***p<0.001 vs CTR; °°°p<0.001 vs <i>Pc</i>AS for each time point.</p

Disease course in mice infected with <i>Pb</i>NK65 or <i>Pc</i>AS.

<p>C57BL/6J mice were injected intraperitoneally with 10⁴ erythrocytes infected with <i>Plasmodium berghei (Pb</i>NK65) or <i>P</i>. <i>chabaudi</i> AS (<i>Pc</i>AS). Peripheral parasitemia (panel A) and weights of the left lungs (panel B) were determined at day 6, 8 and 10 post infection (n = 6–8 mice for each time point and strain). (panel C) Representative pictures of right lungs (not perfused) from uninfected (CTR) or infected mice (day 10 post infection). (panel D) Hz content in lung tissue (pmole Hz /mg lung tissue) at day 10 post infection. *p<0.05; **p<0.01; *** p<0.0001 vs CTR; °°°p< 0.01 <i>Pb</i>NK65 vs <i>Pc</i>AS.(n = 6–8 mice for each time point and strain).</p

Alteration in the lipid profile of plasma from uninfected, <i>Pb</i>NK65 or <i>Pc</i>AS infected mice.

<p>PL (panel A) and neutral lipid content (panel B) of plasma of infected and uninfected mice at day 10 post infection were determinated as described in Materials and Methods.Cho = Cholesterol, ChoE = Cholesterol esters, TG = Triacylglycerols. *p<0.05; **p<0.01 vs CTR; °p<0.01 <i>Pb</i>NK65 vs <i>Pc</i>AS. n = 8–12.</p

Hemozoin Induces Hepatic Inflammation in Mice and Is Differentially Associated with Liver Pathology Depending on the <i>Plasmodium</i> Strain

<div><p>Malaria is a global disease that clinically affects more than two hundred million people annually. Despite the availability of effective antimalarials, mortality rates associated with severe complications are high. Hepatopathy is frequently observed in patients with severe malarial disease and its pathogenesis is poorly understood. Previously, we observed high amounts of hemozoin or malaria pigment in livers from infected mice. In this study, we investigated whether hemozoin is associated with liver injury in different mouse malaria models. C57BL/6J mice infected with the rodent parasites <i>Plasmodium berghei</i> ANKA, <i>P. berghei</i> NK65 or <i>P. chabaudi</i> AS had elevated serum liver enzymes without severe histological changes in the liver, in line with the observations in most patients. Furthermore, liver enzymes were significantly higher in serum of <i>P. chabaudi</i> AS-infected mice compared to mice infected with the <i>P. berghei</i> parasite strains and a strong positive correlation was found between hepatic hemozoin levels, hepatocyte damage and inflammation in the liver with <i>P. chabaudi</i> AS. The observed liver injury was only marginally influenced by the genetic background of the host, since similar serum liver enzyme levels were measured in infected C57BL/6J and BALB/c mice. Intravenous injection of <i>P. falciparum</i>-derived hemozoin in malaria-free C57BL/6J mice induced inflammatory gene transcription in the liver, suggesting that hemozoin may be involved in the pathogenesis of malaria hepatopathy by inducing inflammation.</p></div

Massive inflammatory infiltrates in livers of <i>PcAS</i>-infected mice.

<p>Paraffin-embedded sections were prepared from livers of uninfected C57BL/6J mice (Con) and from mice infected with <i>Pb</i>ANKA (day 8), <i>Pb</i>NK65 (day 10) or <i>Pc</i>AS (day 10), and stained with hematoxylin–eosin. Representative images are shown (scale bars, 50 µm).</p

Hepatic mRNA expression of inflammatory mediators 6 h after intravenous injection of <i>Pf</i>Hz.

<p>Median values are shown for each group (n = 3–12) and min-max values are indicated between parentheses. Data were stratified according to the amount of <i>Pf</i>Hz in the liver (Li): <70 or >200 nmol <i>Pf</i>Hz; Statistically different from PBS administered mice:</p><p>* <i>p</i><0.05;</p><p>** <i>p</i><0.01;</p><p>*** <i>p</i><0.001;</p><p>Statistically different from <i>Pf</i>Hz Li <70:</p>†<p><i>p</i><0.05;</p>††<p><i>p</i><0.01;</p><p>FasL, Fas ligand; Hmox1, heme oxygenase 1; ICAM-1, intercellular adhesion molecule-1; IFN-γ, interferon-gamma; IL-, interleukin-; iNOS, inducible nitric oxide synthase; IP-10, interferon gamma inducible protein-10; KC, keratinocyte-derived chemokine; LT-α, lymphotoxin-alpha; MCP-1, monocyte chemotactic protein-1; MHC-II, major histocompatibility complex class II; MMP-9, matrix metalloproteinase-9/gelatinase B; NOX2, NADPH oxidase 2; TGF-β, transforming growth factor-beta; TNF, tumor necrosis factor.</p><p>Hepatic mRNA expression of inflammatory mediators 6 h after intravenous injection of <i>Pf</i>Hz.</p