Transcription profiling of mouse monocytes recruited in the peritoneum during experimental infection with Listeria monocytogenes

ABSTRACT: The immune system cellular response to tissue damage and infection requires the recruitment of blood leukocytes to the target tissue. This process is mediated through a classical multistep mechanism which involves transient rolling on the endothelium and recognition of inflammation followed by extravasation. We show here, by direct examination of blood monocyte functions in vivo, that resident monocytes monitor the endothelium of healthy tissues through patrolling, a new mechanism which allows extravasation in the absence of rolling.Patrolling depends on the integrin LFA1 and the chemokine receptor CX3CR1, and is required for rapid tissue invasion and initiation of an early immune response by monocytes that differentiate into macrophages at the site of tissue damage and infection. The main goal of the experiment was to compare expression levels of genes in Gr1- monocytes in the blood and after recruitment in the peritoneum during experimental infection with Listeria monocytogenes (time course analysis). Experiment Overall Design: Six-weeks old (C57Bl6, Cx3cr1gfp/+) mice were intraperitonealy infected with a low number (1.104) of L. monocytogenes (EGDe strain) in exponential growth phase (bacteria were grown in BHI at 108/ml, and diluted 10.000x in PBS immediately before injection). Group of three mice were euthanized, before infection (time â??0â??) and 2 and 8 hours after infection (time â??2â?? and â??8â??). Peripheral blood cells were recovered at time â??0â??, and peritoneal cells were recovered at time â??0â??, â??2hâ??, and â??8hâ?? by peritoneal lavage. Cells from individual mice were stained with antibodies to CD11b (PECy7), Gr1 (APC), NK1.1, B220 and CD3 (PE), and F4/80 (biotin-conjugated followed by streptavidinâ??pacific blue) for sorting. Gr1- monocytes were purified as NK1.1- CD3- B220- CD11b+ F4/80low Gr1-, gfphigh; Gr1+ monocytes were purified as NK1.1- CD3- B220- CD11b+ F4/80low Gr1+, gfpint; and polymorphonuclear cells were purified as NK1.1- CD3- B220- CD11b+ F4/80- Gr1high, gfp-. 1.103 cells from each mice, time point, and phenotype were purified by facs sorting according to their phenotype. Samples were kept at 4°C before and during the sort. Cells were directly sorted in the SuperAmp Lysis Buffer (Miltenyi Biotec, Bergisch Gladbach, Germany) using a FACS Aria cell-sorter (BD biosciences).

ORGANISM(S): Mus musculus

SUBMITTER: Silvia Rueberg 

PROVIDER: E-GEOD-8294 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress