Streptococcus suis (SS) is a zoonotic pathogen that can cause meningitis in humans and pigs. The gastro-intestinal tract is an entry site for SS in both pig and human infection. The capsular polysaccharide (CPS), which determines the 33 serotypes reported so far, appears to play an essential role during the interaction of SS with the host mucosa epithelium. Our recent finding showed that in both in vitro and in vivo models SS was able to adhere to and translocate across the porcine and human intestinal epithelial cell monolayers. However, the precise mechanism of adhesion and translocation of SS across Intestinal Epithelium Cells (IEC) is still unknown. SS serotype 2 (SS2) is responsible for most reported infections in humans and pigs worldwide, whilst SS serotype 9 (SS9) is the most common serotype isolated from diseased pigs, but it has never been isolated from human patients. The SS adhesion to and the translocation across human and porcine IEC is associated with SS serotype and genotype.
The aim of this project is to study the bacterial genome-wide transcription during the interactions of SS2 (zoonotic) and SS9 (no-zoonotic) with human and porcine intestinal mucosa to identify the bacterial factors involved in SS-host IEC interaction. We will use whole transcriptome sequencing (RNA-seq) techniques to quantitatively catalog the transcriptome of zoonotic SS and non-zoonotic SS strains derived during human and pig IEC infection using a comparative experimental design.
- Use of in vitro cell culture model of human and porcine IEC
- Co-culture of SS2 strain and SS9 strain with human and porcine IEC
- Optimization of RNA extraction and enrichment of bacterial RNA
- Library preparation (fragmentation, labelling, retrotranscription) to perform SOLiD Sequencing at the Department of Genome Analysis of AMC
- Transcriptome data analysis: The RNA profiling data will be adjusted to permit a comparison between bacterial gene expression
- The changes in metabolic pathways during the SS interaction with intestinal mucosa will be analyzed by SimPhenyTM Software
The student(s) will get a research project from at least one of the objectives above and can participate in discussions of the medical microbiology and meningitis group meetings.
Techniques and procedures you can get acquainted with:
- culture microbes, safely handling pathogens
- microbial RNA extraction and enrichment, preparation of libraries for RNA-Seq, transcriptome data analysis
- cell-based assays to characterise bacterial binding to host cells
from February 2015; preferably 4-6 months
HMI: Peter van Baarlen: Peter.firstname.lastname@example.org
Dept. of Medical Microbiology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands