IFNγ dependent Induction of NOS2/iNOS/Nitrate production in Lymphoid stromal cells.

<p>(<b>a</b>) Costimulatory marker expression on the surface of FRC.5 cells following IFN<b>γ</b> treatment. (<b>b</b>) CFSE dilution profiles of OTI T cells stimulated with peptide-loaded BMDCs in the presence or absence of FRC.5 cells and blocking antibodies to CD80, CD86, or PDL-1. (<b>c</b>) Real-time PCR expression of <i>nos2</i> mRNA in FRC.5 cells cultured with either IFN<b>γ</b>, an iNOS-specific enzyme inhibitor (1400W), or both. (<b>d</b>) Immunofluorescence staining of NOS2 protein in FRC.5 cells cultured with pre-labeled SL8-pulsed BMDC or in the presence of pre-labeled SL8-pulsed BMDC plus pre-labeled T cells. Teal lines around FRC in FRC/DC condition indicate the border of the FRC and white arrowhead indicates a T-DC couple. Scalebar 20 µm. (<b>e</b>) Intracellular flow cytometry for NOS2 protein expression by FRC.5 cells cultured with SL8-pulsed BMDC with or without T cells and with or without IFN<b>γ</b> blockade for 48 hours. (<b>f</b>) Extracellular nitrite concentration measured by Griess assay in FRC.5 cultures either alone; with OTI T cells and peptide-loaded BMDCs; or OTI cells, peptide-loaded BMDCs, and 1400W.