Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes-4

<p><b>Copyright information:</b></p><p>Taken from "Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes"</p><p>http://www.malariajournal.com/content/6/1/73</p><p>Malaria Journal 2007;6():73-73.</p><p>Published online 2 Jun 2007</p><p>PMCID:PMC1904226.</p><p></p>ix (MIX+LPS) containing IFN-gamma (400 U/mL), TNF-alpha (500 U/mL), IL-1beta (100 U/mL) and LPS (20 μg/mL) (final concentrations). After 24 h incubation, mRNA was extracted from cells and iNOS-specific mRNA was quantified by real-time RT-PCR. Data are presented as fold increase in iNOS-mRNA level vs control. Mean values ± SE (n = 2–7). The difference between untreated/unfed controls and treated/fed cells was tested for significance: *p < 0.05

Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes-6

<p><b>Copyright information:</b></p><p>Taken from "Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes"</p><p>http://www.malariajournal.com/content/6/1/73</p><p>Malaria Journal 2007;6():73-73.</p><p>Published online 2 Jun 2007</p><p>PMCID:PMC1904226.</p><p></p>d probed with anti-JAK-2 in Western blot. One representative experiments out of 3 with similar results

Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes-2

<p><b>Copyright information:</b></p><p>Taken from "Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes"</p><p>http://www.malariajournal.com/content/6/1/73</p><p>Malaria Journal 2007;6():73-73.</p><p>Published online 2 Jun 2007</p><p>PMCID:PMC1904226.</p><p></p>ix (MIX+LPS) containing IFN-gamma (400 U/mL), TNF-alpha (500 U/mL), IL-1beta (100 U/mL) and LPS (20 μg/mL) (final concentrations). After 24 h incubation, NO production was measured as nitrite in the cell culture supernatants and expressed as nmole/mg cell protein. Mean values ± SE (n = 2–11). The difference between untreated/unfed controls and treated/fed cells was tested for significance:*p < 0.05; **p < 0.1

Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes-1

<p><b>Copyright information:</b></p><p>Taken from "Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes"</p><p>http://www.malariajournal.com/content/6/1/73</p><p>Malaria Journal 2007;6():73-73.</p><p>Published online 2 Jun 2007</p><p>PMCID:PMC1904226.</p><p></p>d probed with anti-JAK-2 in Western blot. One representative experiments out of 3 with similar results

Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes-0

<p><b>Copyright information:</b></p><p>Taken from "Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes"</p><p>http://www.malariajournal.com/content/6/1/73</p><p>Malaria Journal 2007;6():73-73.</p><p>Published online 2 Jun 2007</p><p>PMCID:PMC1904226.</p><p></p>s (solid thick line) measured by flow cytometry. Background is plotted as solid area

Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes-3

<p><b>Copyright information:</b></p><p>Taken from "Malarial pigment haemozoin, IFN-gamma, TNF-alpha, IL-1beta and LPS do not stimulate expression of inducible nitric oxide synthase and production of nitric oxide in immuno-purified human monocytes"</p><p>http://www.malariajournal.com/content/6/1/73</p><p>Malaria Journal 2007;6():73-73.</p><p>Published online 2 Jun 2007</p><p>PMCID:PMC1904226.</p><p></p>alpha (500 U/mL), IL-1beta (100 U/mL) and LPS (20 μg/mL) (final concentrations). After 24 h incubation cells were analysed for iNOS expression. Immune-precipitated iNOS from lysate proteins of human monocytes (A) or lysate proteins of RAW264.7 murine macrophages (B) were separated by 10% SDS-PAGE and blotted to PVDF. iNOS was visualized via ECL by binding of a monoclonal anti-iNOS antibody. Recombinant iNOS (Sigma) was used as positive control. Blots shown are representative for five separate experiments

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

Spearman correlations between Hz, liver enzymes and mRNA expression levels of inflammation-associated proteins.

<p>* <i>p</i><0.05;</p><p>** <i>p</i><0.01;</p><p>*** <i>p</i><0.001;</p><p>**** <i>p</i><0.0001;</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>Spearman correlations between Hz, liver enzymes and mRNA expression levels of inflammation-associated proteins.</p

Liver pathology is more severe in <i>Pc</i>AS-infected mice than in <i>P. berghei</i>-infected mice.

<p>C57BL/6J mice were infected with <i>Pb</i>ANKA, <i>Pb</i>NK65 or <i>Pc</i>AS and sacrificed at different time points after infection. <i>Pb</i>ANKA-infected mice die around day 8, thus no data are available on day 10. (A) Weights of perfused livers. Means are indicated ± SEM (n = 8–21 for each condition). (B) Peripheral parasitemia levels. Means are indicated ± SEM (n = 8–21 for each group). (C–E) Serum levels of ALT (panel C), AST (panel D) and GLDH (panel E) levels. Data are pooled from two separate experiments with similar results. Each dot represents the results from an individual mouse. Small horizontal stripes between individual data points represent group medians and horizontal lines with asterisks on top indicate statistical differences between groups. Asterisks above individual data sets indicate statistical differences with the uninfected control group as follows: * <i>p</i><0.05, ** <i>p</i><0.01 and *** <i>p</i><0.001.</p

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