The effect of brain co-culture on the protein expression of skin-derived and glandular stem cells.

<p>Co-cultivation with rat brain biopsies led to comparable changes in the expression of some structure proteins in all three human stem cell populations. In the case of Nestin-expressing cells a distinctive decrease by up to one third could be detected, with the slight exception of the SDSC (<b>A–F</b>). The percentage of NF-positive cells increased in all populations up to 4-fold, accompanied by the general exhibition of an elongated morphology resembling uni- and bipolar neurons (<b>G–L</b>; arrows). α-SMA-positive cells decreased in number and/or showed a visible degradation of the actin filaments (<b>M–R</b>). Nuclei were counterstained with DAPI (blue). Blue bars represent the total cell count (via DAPI) in each image, red and green bars the relative amount of positively stained cells.</p

Detection of secreted growth factors via microarray.

<p>Supernatants of co-cultures and controls were tested for the presence of growth factors via a microarray-based technique (<b>A</b>). The processed arrays were semi-quantitatively analyzed by fluorescence, whereby representative images for every approach are shown (<b>B</b>). Growth factors that were found in at least four of five experiments were comparatively described in their expression based on the fluorescence signals (blue = weak/+; green-yellow = moderate/++; red-white = strong/+++) (<b>C</b>).</p

Neuron-like development in skin- and gland-derived stem cells during co-culture.

<p>An immunocytochemical staining revealed the co-localization of the neuroprogenitor marker Nestin and the neuronal cell marker Neurofilament in co-culture stimulated SDSC, PDSC and PSC (<b>A</b>). Furthermore, the establishment of axon-like cell processes with lengths up to 450 µm could be detected after co-cultivation (<b>B</b>).</p

Immunocytochemical characterization of skin- and gland-derived stem cells concerning their stemness.

<p>SDSC, PDSC and PSC generally express the stem cell markers Nestin, Sox 2 and Oct 4 as well as marker proteins for protein synthesis (Vigilin) and proliferation (Ki67). Nuclei were counterstained with DAPI (blue).</p

Immunocytochemical characterization of skin- and gland-derived stem cells concerning their differentiation capability.

<p>The used stem cell lines (SDSC, PDSC, PSC) spontaneously differentiate <i>in vitro</i> and, in the course of this differentiation, express marker proteins for the ectodermal (NF, GFAP, CK18, panCK) and mesodermal (α-SMA) germ layer. Nuclei were counterstained with DAPI (blue).</p

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<p>Anti-neutrophil cytoplasmic autoantibodies (ANCA) targeting proteinase 3 (PR3) and myeloperoxidase expressed by innate immune cells (neutrophils and monocytes) are salient diagnostic and pathogenic features of small vessel vasculitis, comprising granulomatosis with polyangiitis (GPA), microscopic polyangiitis, and eosinophilic GPA. Genetic studies suggest that ANCA-associated vasculitides (AAV) constitute separate diseases, which share common immunological and pathological features, but are otherwise heterogeneous. The successful therapeutic use of anti-CD20 antibodies emphasizes the prominent role of ANCA and possibly other autoantibodies in the pathogenesis of AAV. However, to elucidate causal effects in AAV, a better understanding of the complex interplay leading to the emergence of B lymphocytes that produce pathogenic ANCA remains a challenge. Different scenarios seem possible; e.g., the break of tolerance induced by a shift from non-pathogenic toward pathogenic autoantigen epitopes in inflamed tissue. This review gives a brief overview on current knowledge about genetic and epigenetic factors, barrier dysfunction and chronic non-resolving inflammation, necro-inflammatory auto-amplification of cellular death and inflammation, altered autoantigen presentation, alternative complement pathway activation, alterations within peripheral and inflamed tissue-residing T- and B-cell populations, ectopic lymphoid tissue neoformation, the characterization of PR3-specific T-cells, properties of ANCA, links between autoimmune disease and infection-triggered pathology, and animal models in AAV.</p