Figure 1
From: Stromal cells as trend-setters for cells migrating into the lymph node
Isolated stromal cells (SCs) from different lymph nodes express different levels of cytokines. CD45− SCs from mesenteric LN (mLN) and peripheral LN (pLN) were isolated, and flow cytometry, mRNA expression microarray analysis, real-time PCR, and protein detection were performed. (a) Various subpopulations of SCs were identified within CD45− cells with a purity of ∼98%. Using a combination of anti-CD31 and anti-gp38 antibodies blood endothelial cells (BECs), lymph endothelial cells (LEC), and fibroblastic reticular cells (FRCs) were recognized. Follicular dendritic cells (FDCs) were detected by the anti-FDC-M2 antibody, gated from the CD31/gp38 double-negative (DN) population. (b) Genes encoding interleukins or chemokines that fulfilled the applied filter criteria for different mRNA expressions in mLN vs. pLN SCs are depicted. Fold difference value (pLN/mLN) was calculated from normalized processed signal intensities (nPS) and depicted for both independent experiments performed. Agilent Probe-IDs, accession numbers, and gene symbols are given. In cases in which more than one probe directed against the same transcript were present on the microarray and fulfilled the applied filter criteria, one representative probe measurement was selected for visualization. (c) The cytokine and mRNA levels produced by SCs were determined using a bio-plex analysis and real-time PCR, respectively. SCs were isolated and cultured for 24 h. Supernatants and cells were collected and analyzed (n=3–6). Significant differences in the unpaired t-test are indicated by *P<0.05; **P<0.01.
