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    ABSTRACT: Autoimmune hepatitis (AIH) is an important cause of severe liver disease and is associated with both quantitative and qualitative regulatory T cell (Treg) impairments. We have shown that Tregs express CD39, an ectonucleotidase responsible for extracellular nucleotide hydrolysis, culminating in the production of immunosuppressive adenosine. In this study, we describe multiple CD39(pos) Treg defects that potentially contribute to the impaired immuno-regulation that is characteristic of AIH. We have examined the frequency and phenotype of CD39(pos) Tregs by flow cytometry and measured their ectonucleotidase activity. The capacity of CD4(pos) CD25(high) , CD4(pos) CD25(high) CD39(pos) and CD4(pos) CD25(high) CD39(neg) subsets to suppress both proliferation of effector T cells and IL17 production was evaluated. In AIH, CD39(pos) Tregs are decreased in frequency, exhibit limited adenosine triphosphate (ATP)/adenosine diphosphate (ADP) hydrolysis activity and fail to suppress IL17 production by effector CD4 T cells. Moreover, these CD39(pos) Tregs display a more pro-inflammatory profile in AIH, which is characterised by elevated CD127 positivity, and a greater propensity to produce IFNγ or IL17 upon challenge with pro-inflammatory stimuli. In AIH CD39(pos) Tregs are decreased in number, fail to adequately hydrolyse pro-inflammatory nucleotides and do not suppress efficiently IL17 production by effector CD4 T cells. CD39(pos) Tregs show plasticity and are unstable upon pro-inflammatory challenge, suggesting that defective immuno-regulation in AIH might result not only from reduced Treg number and function but also from increased conversion of Tregs into effector cells. (HEPATOLOGY 2013.).
    Full-text · Article · Mar 2014 · Hepatology
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    ABSTRACT: CD46 is an important regulator of the complement system by preventing unwanted deposition of the complement activation products and opsonins C3b/C4b onto self-tissue. Recently, intracellular signals mediated by CD46 activation on several distinct human cell types have demonstrated that CD46 also plays decisive roles in immuneregulation. The growing recognition of CD46 as key regulator in several vital biological processes, led to increased demand in sensitive methods for monitoring CD46 expression and changes thereof on cells and in tissues. Here we describe a method, which allows for studying CD46 expression on the surface of cells using specific antibodies in combination with fluorescence-activated cell sorting (FACS) analysis.
    No preview · Article · Jan 2014 · Methods in molecular biology (Clifton, N.J.)
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    ABSTRACT: CD59 is the single regulator of the terminal complement pathway. It has been implicated in disease such as Paroxysmal nocturnal hemoglobinuria (PMH) and cancer. Expression of CD59 protects normal and malignant cells from the cytotoxic potential of the complement system. Here we describe a method, which allows for studying its expression on the surface of cells.
    No preview · Article · Jan 2014 · Methods in molecular biology (Clifton, N.J.)
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