Figure 1

A. The data show increased expression of known immune suppressive factors by TLR3-primed hMSCs (MSC2) but not by TLR4-primed hMSCs (MSC1). Methods: MSCs were pre-treated for 1 hr with TLR agonists (LPS for MSC1 or poly(I∶C) for MSC2), washed and cultured for an additional 48 hr prior to harvesting the spent medium and analysis with Bio-Plex Cytokine Assays (Human Group I & II; Bio-Rad, Hercules, CA) following the manufacturer's instructions. Data are presented by the quantitative comparative CT (threshold value) method [46]. Error bars indicate SEM. Data are representative of triplicate measurements with 5 MSC donors. B. The data implicate direct TLR3 induction of IP10 (CCL10) and RANTES (CCL5) secretion. Methods: hMSCs were transfected with pZERO-hTLR3 and pZERO-hTLR4 (Invivogen), using 250 ng plasmid/1×10⁶ cells (nucleofector). Cells from each transfection were left untreated or stimulated with TLR3 and TLR4 agonists for 1 hr washed and incubated for 48 hr. Conditioned medium was harvested and analyzed as in A. Transfection efficiency was determined by these methods to be 30–35%. Data are representative of triplicate measurements with 3 MSC donors.