TY  - JOUR
A1  - Tylek, Tina
A1  - Blum, Carina
A1  - Hrynevich, Andrei
A1  - Schlegelmilch, Katrin
A1  - Schilling, Tatjana
A1  - Dalton, Paul D
A1  - Groll, Jürgen
T1  - Precisely defined fiber scaffolds with 40 μm porosity induce elongation driven M2-like polarization of human macrophages
JF  - Biofabrication
N2  - Macrophages are key players of the innate immune system that can roughly be divided into the pro-inflammatory M1 type and the anti-inflammatory, pro-healing M2 type. While a transient initial pro-inflammatory state is helpful, a prolonged inflammation deteriorates a proper healing and subsequent regeneration. One promising strategy to drive macrophage polarization by biomaterials is precise control over biomaterial geometry. For regenerative approaches, it is of particular interest to identify geometrical parameters that direct human macrophage polarization. For this purpose, we advanced melt electrowriting (MEW) towards the fabrication of fibrous scaffolds with box-shaped pores and precise inter-fiber spacing from 100 μm down to only 40 μm. These scaffolds facilitate primary human macrophage elongation accompanied by differentiation towards the M2 type, which was most pronounced for the smallest pore size of 40 μm. These new findings can be important in helping to design new biomaterials with an enhanced positive impact on tissue regeneration.
KW  - cell elongation
KW  - human macrophages
KW  - melt electrowriting (MEW)
KW  - macrophage polarization
KW  - 3D scaffolds
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-254012
VL  - 12
IS  - 2
ER  - 
TY  - JOUR
A1  - Afonso-Grunz, Fabian
A1  - Hoffmeier, Klaus
A1  - Müller, Sören
A1  - Westermann, Alexander J.
A1  - Rotter, Björn
A1  - Vogel, Jörg
A1  - Winter, Peter
A1  - Kahl, Günter
T1  - Dual 3'Seq using deepSuperSAGE uncovers transcriptomes of interacting Salmonella enterica Typhimurium and human host cells
JF  - BMC Genomics
N2  - Background: 
The interaction of eukaryotic host and prokaryotic pathogen cells is linked to specific changes in the cellular proteome, and consequently to infection-related gene expression patterns of the involved cells. To simultaneously assess the transcriptomes of both organisms during their interaction we developed dual 3'Seq, a tag-based sequencing protocol that allows for exact quantification of differentially expressed transcripts in interacting pro-and eukaryotic cells without prior fixation or physical disruption of the interaction. 

Results: 
Human epithelial cells were infected with Salmonella enterica Typhimurium as a model system for invasion of the intestinal epithelium, and the transcriptional response of the infected host cells together with the differential expression of invading and intracellular pathogen cells was determined by dual 3'Seq coupled with the next-generation sequencing-based transcriptome profiling technique deepSuperSAGE (deep Serial Analysis of Gene Expression). Annotation to reference transcriptomes comprising the operon structure of the employed S. enterica Typhimurium strain allowed for in silico separation of the interacting cells including quantification of polycistronic RNAs. Eighty-nine percent of the known loci are found to be transcribed in prokaryotic cells prior or subsequent to infection of the host, while 75% of all protein-coding loci are represented in the polyadenylated transcriptomes of human host cells.

Conclusions: 
Dual 3'Seq was alternatively coupled to MACE (Massive Analysis of cDNA ends) to assess the advantages and drawbacks of a library preparation procedure that allows for sequencing of longer fragments. Additionally, the identified expression patterns of both organisms were validated by qRT-PCR using three independent biological replicates, which confirmed that RELB along with NFKB1 and NFKB2 are involved in the initial immune response of epithelial cells after infection with S. enterica Typhimurium.
KW  - complete genome sequence
KW  - secretion systems
KW  - RNA-Seq
KW  - deepSuperSAGE
KW  - transcriptome
KW  - gene expression
KW  - serovar Typhimurium
KW  - human macrophages
KW  - epithelial cells
KW  - infection
KW  - SuperSAGE
KW  - receptors
KW  - Dual 3'seq
KW  - MACE
KW  - tag based
KW  - simultaneous
KW  - genome wide
KW  - gene expression profiling
KW  - host pathogen interaction
KW  - Salmonella enterica Typhimurium strain SL1344
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143230
VL  - 16
IS  - 323
ER  -