It could help generating a proper immune response against Giardia

It could help generating a proper immune response against Giardia and inhibiting pathophysiological effects in the intestinal epithelium that are caused by arginine-consumption of Giardia. Conclusion The findings S6 Kinase inhibitor presented here, and earlier data, clearly show that Giardia interferes with a proper host immune response

of the host intestinal epithelium on the innate and adaptive immunity level by affecting arginine in the intesine on multiple levels (Figure 1). The parasite consumes arginine as an energy source [7, 24] and thereby the substrate for NOS [10]. Giardia trophozoites release arginine-consuming enzymes ADI and OCT [9] and ornithine that blocks the host cell transporter for arginine uptake [29]. Expression of iNOS is initially induced but selleck inhibitor reduced by the parasite at later stages of infection. Expression of ODC is also induced, further shifting arginine-flux away from iNOS. Flavohemoglobin expression is induced in Giardia early upon NO-stress [13]. Dendritic cell cytokine production [22] and T cell proliferation is affected

due to reduced arginine-availability. All the observed effects might not be overwhelmingly strong by themselves, but the sum of them will certainly protect the parasite from the host’s response. Methods Ethics statement Individuals contributing peripheral blood mononuclear cells (PBMC) for the study of T cell proliferation gave written consent in a standard form upon registration as blood donors. The study and consent procedure was approved by the Regional

Committee for Ethics in Medical Research (REK), Bergen, triclocarban Norway. Reagents this website and cell culture If not stated otherwise, all chemicals and reagents were purchased from Sigma Chemical Co, USA. G. intestinalis trophozoites (strain WB, clone C6 (ATCC30957), strain GS, clone H7 (ATCC50581) and strain P15 were maintained in Giardia growth medium, TYDK, as described in Stadelmann et al [7]. G. intestinalis trophozoites were used for interaction with human intestinal epithelial cells (IECs) when reaching confluence. They were washed in PBS twice and counted before dilution in complete DMEM (high-glucose DMEM with 10% fetal bovine serum (Gibco®, Invitrogen, Paisley, UK), 4 mM L-glutamine, 1 × MEM non-essential amino acids, 160 μg/mL streptomycin and 160 U/mL penicillin G) and addition to IECs at indicated numbers. IEC cell lines CaCo-2, clone TC7 and HCT-8 (ATCC CCL-244), were maintained as described in Stadelmann et al [2, 7], at 37°C, 5% CO2, in humid atmosphere, the same conditions that were applied for interaction experiments. Giardia – IEC interaction: gene expression For assessment of gene expression of G. intestinalis infected human IECs, Caco-2 cells were cultured in T25 tissue culture flasks 21 days post confluence with medium changes twice per week to allow differentiation [9].

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