TY  - JOUR
A1  - Heydarian, Motaharehsadat
A1  - Schweinlin, Matthias
A1  - Schwarz, Thomas
A1  - Rawal, Ravisha
A1  - Walles, Heike
A1  - Metzger, Marco
A1  - Rudel, Thomas
A1  - Kozjak-Pavlovic, Vera
T1  - Triple co-culture and perfusion bioreactor for studying the interaction between Neisseria gonorrhoeae and neutrophils: A novel 3D tissue model for bacterial infection and immunity
JF  - Journal of Tissue Engineering
N2  - Gonorrhea, a sexually transmitted disease caused by the bacteria Neisseria gonorrhoeae, is characterized by a large number of neutrophils recruited to the site of infection. Therefore, proper modeling of the N. gonorrhoeae interaction with neutrophils is very important for investigating and understanding the mechanisms that gonococci use to evade the immune response. We have used a combination of a unique human 3D tissue model together with a dynamic culture system to study neutrophil transmigration to the site of N. gonorrhoeae infection. The triple co-culture model consisted of epithelial cells (T84 human colorectal carcinoma cells), human primary dermal fibroblasts, and human umbilical vein endothelial cells on a biological scaffold (SIS). After the infection of the tissue model with N. gonorrhoeae, we introduced primary human neutrophils to the endothelial side of the model using a perfusion-based bioreactor system. By this approach, we were able to demonstrate the activation and transmigration of neutrophils across the 3D tissue model and their recruitment to the site of infection. In summary, the triple co-culture model supplemented by neutrophils represents a promising tool for investigating N. gonorrhoeae and other bacterial infections and interactions with the innate immunity cells under conditions closely resembling the native tissue environment.
KW  - Triple co-culture
KW  - biomimetic 3D tissue model
KW  - Neisseria gonorrhoeae
KW  - perfusion-based bioreactor system
KW  - neutrophil transmigration
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259032
VL  - 12
ER  - 
TY  - JOUR
A1  - Kunz, Tobias C.
A1  - Rühling, Marcel
A1  - Moldovan, Adriana
A1  - Paprotka, Kerstin
A1  - Kozjak-Pavlovic, Vera
A1  - Rudel, Thomas
A1  - Fraunholz, Martin
T1  - The Expandables: Cracking the Staphylococcal Cell Wall for Expansion Microscopy
JF  - Frontiers in Cellular and Infection Microbiology
N2  - Expansion Microscopy (ExM) is a novel tool improving the resolution of fluorescence microscopy by linking the sample into a hydrogel that gets physically expanded in water. Previously, we have used ExM to visualize the intracellular Gram-negative pathogens Chlamydia trachomatis, Simkania negevensis, and Neisseria gonorrhoeae. Gram-positive bacteria have a rigid and thick cell wall that impedes classic expansion strategies. Here we developed an approach, which included a series of enzymatic treatments resulting in isotropic 4× expansion of the Gram-positive pathogen Staphylococcus aureus. We further demonstrate the suitability of the technique for imaging of planktonic bacteria as well as endocytosed, intracellular bacteria at a spatial resolution of approximately 60 nm with conventional confocal laser scanning microscopy.
KW  - high-resolution imaging
KW  - endosomes
KW  - autophagosomes
KW  - host-pathogen interaction
KW  - expansion microscopy
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232292
SN  - 2235-2988
VL  - 11
ER  -