Na&iuml;ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

Over the last decade, several adult stem cell  populations have been identified in human skin 1-4.  The isolation of multipotent adult dermal precursors ...

Technische Universität München

T cells exercise their full impact on target cells through a combination of secreted cytokines. The recently described T helper cell subset Th22 is characterized by a combinatorial secretion of IL-22 and TNF-Î±. Here, we demonstrate that IL-22 increases the TNF-Î±-dependent induction and secretion of several immune-modulatory molecules such as initial complement factors C1r and C1s, antimicrobial peptides S100A7 and HBD-2 (human Î² defensin 2), and antimicrobial chemokines CXCL-9/-10/-11 in primary human keratinocytes. The synergism of IL-22 and TNF-Î± is transmitted intracellularly by MAP kinases and downstream by transcription factors of the AP-1 family. The induction of innate immunity is relevant in an in vitro infection model, where keratinocytes stimulated with Th22 supernatants or recombinant IL-22 plus TNF-Î± effectively inhibit the growth of Candida albicans and maintain survival of epithelia. Accordingly, the combinatorial stimulation of keratinocytes with IL-22 and TNF-Î± most efficiently conserves the integrity of the epidermal barrier in a three-dimensional skin infection model as compared with IFN-Î³, IL-17, IL-22 or TNF-Î± alone. In summary, we demonstrate that IL-22 and TNF-Î± represent a potent, synergistic cytokine combination for cutaneous immunity.

IL-22 and TNF-Î± represent a key cytokine combination for epidermal integrity during infection with Candida albicans.

Cellules T exercent leur impact sur les cellules cibles gr&acirc;ce &agrave; une combinaison de cytokines s&eacute;cr&eacute;t&eacute;es. Le sous-ensemble de cellules d&#39;assistance T r&eacute;cemment d&eacute;crit Th22 est caract&eacute;ris&eacute; par une s&eacute;cr&eacute;tion combinatoire de TNF-&alpha; et IL-22. Ici, nous d&eacute;montrons qu&#39;IL-22 augmente l&#39;induction d&eacute;pendante de TNF-&alpha; et s&eacute;cr&eacute;tion de plusieurs mol&eacute;cules modulatrices-immunitaires tels que les facteurs du compl&eacute;ment initial C1r et C1s, peptides antimicrobiens S100A7 et DBC-2 (&beta; humaine defensin 2) et chimiokines antimicrobien CXCL-9 /-10 /-11 dans les k&eacute;ratinocytes humains primaires. Le synergisme de TNF-&alpha; et IL-22 est transmise dans les cellules par des MAP kinases et en aval par des facteurs de transcription de la famille de l&#39;AP-1. L&#39;induction de l&#39;immunit&eacute; inn&eacute;e est pertinente dans un mod&egrave;le d&#39;infection in vitro, o&ugrave; les k&eacute;ratinocytes stimulent avec des surnageants Th22 ou recombinantes IL-22 et TNF-&alpha; inhibent la croissance de Candida albicans et de maintiennent efficacement la survie de l&#39;&eacute;pith&eacute;lium. En cons&eacute;quence, la stimulation combinatoire des k&eacute;ratinocytes avec TNF-&alpha; et IL-22 conserve plus efficacement l&#39;int&eacute;grit&eacute; de la barri&egrave;re &eacute;pidermique dans un mod&egrave;le tridimensionnel de peau d&#39;infection comparativement &agrave; l&#39;IFN-&gamma;, IL-17, IL-22 ou TNF-&alpha; lui seul. En r&eacute;sum&eacute;, nous d&eacute;montrons que le TNF-&alpha; et IL-22 repr&eacute;sentent une combinaison puissante, synergique cytokine pour l&#39;immunit&eacute; cutan&eacute;e.

TNF-&alpha; et IL-22 repr&eacute;sentent une combinaison de cytokine cl&eacute; pour int&eacute;grit&eacute; &eacute;pidermique lors de l&#39;infection &agrave; Candida albicans.

Le cellule T esercitano il loro impatto sulle cellule bersaglio attraverso una combinazione di citochine secrete. Il sottoinsieme di cellule helper T recentemente descritto Th22 &egrave; caratterizzato da una secrezione combinatoria IL 22 e il TNF-&alpha;. Qui, ci dimostrano che IL 22 aumenta l&#39;induzione di TNF-&alpha;-dipendente e secrezione di diverse molecole modulatore immunitaria quali fattori iniziali complemento C1r e C1s, peptidi antimicrobici S100A7 e HBD-2 (&beta; umana defensina 2) e antimicrobica chemochine CXCL-9-10 /-11 nei cheratinociti umani primari. La sinergia di IL-22 e TNF-&alpha; &egrave; trasmessa intracellularly da chinasi mappa e a valle di fattori di trascrizione della famiglia AP-1. L&#39;induzione dell&#39;immunit&agrave; innata &egrave; rilevante in un modello di infezione in vitro, dove cheratinociti stimolati con Th22 supernatanti o IL ricombinante-22 plus TNF-&alpha; efficacemente inibire la crescita di Candida albicans e mantenere la sopravvivenza degli epiteli. Di conseguenza, la stimolazione combinatoria dei cheratinociti con IL 22 e il TNF-&alpha; in modo pi&ugrave; efficiente conserva l&#39;integrit&agrave; della barriera epidermica in un modello di infezione della pelle tridimensionale rispetto a IFN-&gamma;, IL-17, IL-22 o TNF-&alpha; da solo. In sintesi, ci dimostrano che IL 22 e il TNF-&alpha; rappresentano una combinazione sinergica, potente citochina per immunit&agrave; cutanea.

IL 22 e il TNF-&alpha; rappresentano una combinazione di citochina chiave per l&#39;integrit&agrave; dell&#39;epidermide durante l&#39;infezione da Candida albicans.

Hutchinson-Gilford progeria syndrome (HGPS, OMIM 176670) is a rare disorder characterized by segmental accelerated aging and early death from coronary artery disease or stroke. Nearly 90% of HGPS sufferers carry a G608G mutation within exon 11 of LMNA, producing a truncated form of prelamin A, referred to as "progerin". Here, we report the isolation of naÃ¯ve multipotent skin-derived precursor (SKP) cells from dermal fibroblast cultures from HGPS donors. These cells form spheres and express the neural crest marker, nestin, in addition to the multipotent markers, OCT4, Sox2, Nanog and TG30; these cells can self-renew and differentiate into smooth muscle cells (SMCs) and fibroblasts. The SMCs derived from the HGPS-SKPs accumulate nuclear progerin with increasing passages. A subset of the HGPS-naÃ¯ve SKPs express progerin in vitro and in situ in HGPS skin sections. This is the first in vivo evidence that progerin is produced in adult stem cells, and implies that this protein could induce stem cells exhaustion as a mechanism contributing to aging. Our study provides a basis on which to explore therapeutic applications for HGPS stem cells and opens avenues for investigating the pathogenesis of other genetic diseases.

NaÃ¯ve adult stem cells from patients with Hutchinson-Gilford progeria syndrome express low levels of progerin in vivo.

Syndrome de progeria de Hutchinson-Gilford (hormones de croissance, OMIM 176670) est une affection rare caract&eacute;ris&eacute;e par vieillissement acc&eacute;l&eacute;r&eacute; segmentaire et la mort pr&eacute;coce de la maladie coronarienne ou d&#39;accident vasculaire c&eacute;r&eacute;bral. Pr&egrave;s de 90 % des personnes souffrant d&#39;hormones de croissance portent une mutation du g&egrave;ne LMNA, produisant une forme tronqu&eacute;e de l&#39;identification du prelamin A, d&eacute;nomm&eacute; &laquo; progerin &raquo; G608G exon 11. Nous rapportons ici, l&#39;isolement des cellules multipotentes d&eacute;riv&eacute;es de peau pr&eacute;curseur (SKP) na&iuml;ve des cultures de fibroblastes dermiques de donateurs d&#39;hormones de croissance. Ces cellules forment des sph&egrave;res et expriment le marqueur de la cr&ecirc;te neurale, nestin, outre les marqueurs multipotentes, OCT4, Sox2, Nanog et TG30 ; ces cellules peuvent auto-renouveler et se diff&eacute;rencient en cellules musculaires lisses (CML) et de fibroblastes. Le CML d&eacute;riv&eacute; les hormones de croissance-SKP accumule progerin nucl&eacute;aire avec une augmentation de passages. Un sous-ensemble des SKP d&#39;hormones de croissance-na&iuml;ve express progerin in vitro et in situ dans les sections de peau hormones de croissance. Il s&#39;agit de la premi&egrave;re preuve in vivo que progerin est produite dans les cellules souches adultes et implique que cette prot&eacute;ine pouvait induire &eacute;puisement de cellules souches comme un m&eacute;canisme qui contribuent au vieillissement. Notre &eacute;tude fournit une base explorer les applications th&eacute;rapeutiques pour les hormones de croissance des cellules souches et ouvre des pistes pour l&#39;&eacute;tude de la pathogen&egrave;se des autres maladies g&eacute;n&eacute;tiques.

Na&iuml;ve des cellules souches adultes provenant de patients atteints du syndrome de Hutchinson-Gilford progeria expriment des niveaux faibles de progerin in vivo.

Sindrome di progeria di Hutchinson-Gilford (HGPS, OMIM 176670) &egrave; una malattia rara caratterizzata da invecchiamento accelerato segmentale e morte precoce di malattia coronarica o ictus. Quasi il 90% dei malati di HGPS trasportano una mutazione nell&#39;esone 11 G608G LMNA, producendo una forma troncata di prelamin A, indicato come &quot;progerin&quot;. Riportiamo l&#39;isolamento delle cellule multipotenti derivate da pelle precursore (SKP) ingenuo da colture di fibroblasti dermici da donatori HGPS. Queste cellule formano sfere ed esprimono il marcatore cresta neurale, la nestina, oltre i marcatori multipotenti, OCT4, Sox2, Nanog e TG30; Queste cellule possono auto-rinnovarsi e differenziarsi in cellule muscolari lisce (SMC) e fibroblasti. SMCs derivato da HGPS-SKPs si accumulano progerin nucleare con l&#39;aumento di passaggi. Un sottoinsieme dei SKPs HGPS-na&iuml;ve esprimono progerin in vitro ed in situ in HGPS pelle sezioni. Questa &egrave; la prima prova in vivo che progerin &egrave; prodotto in cellule staminali adulte e implica che questa proteina potrebbe indurre cellule staminali esaurimento come un meccanismo che contribuiscono all&#39;invecchiamento. Il nostro studio fornisce una base esplorare le applicazioni terapeutiche per le cellule staminali HGPS e apre le vie per indagare la patogenesi di altre malattie genetiche.

Vera Wenzel

Department of Dermatology and Institute for Medical Engineering

Technische Universität München

Naïve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

Vera Wenzel

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Dermatology and Institute for Medical Engineering

Technische Universität München

Naïve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

Department of Dermatology and Institute for Medical Engineering