IL-1β signaling is critical for regulating CD8⁺ T cell effector activity within the CNS.

Abstract

<p>Examination of T lymphocyte effector activity in the CNS by flow cytometry. Cell were isolated from the brains of mock or day 6–10 p.i. WT (closed circles) or <i>Il-1r^−/−</i>(open squares) and staining for CD3, CD8 and WNV-NS4b tetramer was performed. Quantitation of CD3⁺/CD8+ T lymphocytes (<b>A</b>), CD3⁺/CD8⁺/NS4b⁺ (<b>B</b>) as derived from total cell numbers or frequency of CD3⁺/CD8⁺/NS4b⁺ cells (<b>C</b>). Brains were harvested from day 8 or 10 p.i. and stimulated with 1 µM WNV-NS4b for 4 hrs (<b>D,E, H,I</b>) and assessed for cytokine expression by intracellular staining. Representative dot-plot analysis for IFN-γ and TNF-α expression at day 10 p.i. in the brain (<b>D</b>) or perforin and granzyme B (<b>H</b>). Data are shown as percent cytokine positive for WT (closed box) or <i>Il-1r^−/−</i> (open box) (<b>E,I</b>). Quantification of IFN-γ secretion by ELISA. Total brain cells (2.5e5) were stimulated with NS4b for 24 hrs and IFN-γ was quantified by ELISA for WT (closed circles) and <i>Il-1r^−/−</i> (open squares) cells (<b>F</b>). Data are shown as mean +/− S.E.M. for n = 3–4 mice per time-point (<b>A–E, H,I</b>) or n = 3–7 (<b>F</b>) and are representative of 2–3 independent experiments. *p<0.05, ** p<0.005.</p