TY  - JOUR
A1  - Zadeh-Khorasani, Maryam
A1  - Nolte, Thomas
A1  - Mueller, Thomas D.
A1  - Pechlivanis, Markos
A1  - Rueff, Franziska
A1  - Wollenberg, Andreas
A1  - Fricker, Gert
A1  - Wolf, Eckhard
A1  - Siebeck, Matthias
A1  - Gropp, Roswitha
T1  - NOD-scid IL2R \(\gamma^{null}\) mice engrafted with human peripheral blood mononuclear cells as a model to test therapeutics targeting human signaling pathways
JF  - Journal of Translational Medicine
N2  - Background: Animal models of human inflammatory diseases have limited predictive quality for human clinical trials for various reasons including species specific activation mechanisms and the immunological background of the animals which markedly differs from the genetically heterogeneous and often aged patient population. 
Objective: Development of an animal model allowing for testing therapeutics targeting pathways involved in the development of Atopic Dermatitis (AD) with better translatability to the patient. 
Methods: NOD-scid IL2R \(\gamma^{null}\) mice engrafted with human peripheral blood mononuclear cells (hPBMC) derived from patients suffering from AD and healthy volunteers were treated with IL-4 and the antagonistic IL-4 variant R121/Y124D (Pitrakinra). Levels of human (h) IgE, amount of B-, T- and plasma-cells and ratio of CD4 : CD8 positive cells served as read out for induction and inhibition of cell proliferation and hIgE secretion. Results were compared to in vitro analysis. 
Results: hIgE secretion was induced by IL-4 and inhibited by the IL-4 antagonist Pitrakinra in vivo when formulated with methylcellulose. B-cells proliferated in response to IL-4 in vivo; the effect was abrogated by Pitrakinra. IL-4 shifted CD4 : CD8 ratios in vitro and in vivo when hPBMC derived from healthy volunteers were used. Pitrakinra reversed the effect. Human PBMC derived from patients with AD remained inert and engrafted mice reflected the individual responses observed in vitro. 
Conclusion: NOD-scid IL2R \(\gamma^{null}\) mice engrafted with human PBMC reflect the immunological history of the donors and provide a complementary tool to in vitro studies. Thus, studies in this model might provide data with better translatability from bench to bedside.
KW  - atopic dermatitis
KW  - T-cells
KW  - rheumatoid arthritis
KW  - human interleukin-4
KW  - TGN1412
KW  - oxazolone colitis
KW  - cytokine release
KW  - expression
KW  - antagonists
KW  - responses
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-122960
SN  - 1479-5876
VL  - 11
IS  - 4
ER  - 
TY  - JOUR
A1  - Zhao, Bo
A1  - Zhang, Keya
A1  - Bhuripanyo, Karan
A1  - Choi, Chan Hee J.
A1  - Villhauer, Eric B.
A1  - Li, Heng
A1  - Zheng, Ning
A1  - Kiyokawa, Hiroaki
A1  - Schindelin, Hermann
A1  - Yin, Jun
T1  - Profiling the Cross Reactivity of Ubiquitin with the Nedd8 Activating Enzyme by Phage Display
JF  - PLoS ONE
N2  - The C-terminal peptides of ubiquitin (UB) and UB-like proteins (UBLs) play a key role in their recognition by the specific activating enzymes (E1s) to launch their transfer through the respective enzymatic cascades thus modifying cellular proteins. UB and Nedd8, a UBL regulating the activity of cullin-RING UB ligases, only differ by one residue at their C-termini; yet each has its specific E1 for the activation reaction. It has been reported recently that UAE can cross react with Nedd8 to enable its passage through the UB transfer cascade for protein neddylation. To elucidate differences in UB recognition by UAE and NAE, we carried out phage selection of a UB library with randomized C-terminal sequences based on the catalytic formation of UB similar to NAE thioester conjugates. Our results confirmed the previous finding that residue 72 of UB plays a "gate-keeping" role in E1 selectivity. We also found that diverse sequences flanking residue 72 at the UB C-terminus can be accommodated by NAE for activation. Furthermore heptameric peptides derived from the C-terminal sequences of UB variants selected for NAE activation can function as mimics of Nedd8 to form thioester conjugates with NAE and the downstream E2 enzyme Ubc12 in the Nedd8 transfer cascade. Once the peptides are charged onto the cascade enzymes, the full-length Nedd8 protein is effectively blocked from passing through the cascade for the critical modification of cullin. We have thus identified a new class of inhibitors of protein neddylation based on the profiles of the UB C-terminal sequences recognized by NAE.
KW  - protein NEDD8
KW  - E1
KW  - system
KW  - conjugation
KW  - pathway
KW  - complex
KW  - ligases
KW  - purification
KW  - neddylation
KW  - expression
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-128479
SN  - 1932-6203
VL  - 8
IS  - e70312
ER  -