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Lymphotoxin, but Not TNF, Is Required for Prion Invasion of Lymph Nodes

Figure 4

Vessel-associated MadCam1 expression in lymph nodes is preserved in the absence of TNFR1 signaling.

Frozen sections from mesenteric lymph nodes (mLN) of C57BL/6 (WT) Ig-treated (A), TNFR1−/− Ig-treated (B), C57BL/6 (WT) LTβR-Ig-treated (C), or TNFR1−/− LTβR-Ig-treated (D) mice were analyzed by immunofluorescence with MadCam1 antibody. In WT-Ig mLNs (A), MadCam1 robustly stained thick vessels (yellow arrow), while germinal centers were weakly MadCam1-positive (white arrow). In TNFR1−/−-Ig mLNs (B), MadCam1-positive germinal centers were absent, while vessel-associated MadCam1 staining persisted. In contrast, both WT-LTβR-Ig and TNFR1−/−-LTβR-Ig mLNs were MadCam1-negative (C & D). Size bars = 100 µm. Total mRNA was isolated from spleens (E) or mesenteric lymph nodes (F) of C57BL/6 (WT) Ig-treated, C57BL/6 (WT) LTβR-Ig-treated, TNFR1−/− Ig-treated, or TNFR1−/− LTβR-Ig-treated mice and analyzed for MadCam1 expression by Real Time PCR (Mean ± S.E.M.: Spleen – WT-Ig = 104.21±16.56, WT-LTβR-Ig = 29.98±1.28, TNFR1−/−-Ig = 30.41±2.56, and TNFR1−/−-LTβR-Ig = 27.95±3.77; mLN – WT-Ig = 100.89±8.06, WT-LTβR-Ig = 45.40±6.75, TNFR1−/−-Ig = 49.42±0.63, and TNFR1−/−-LTβR-Ig = 24.79±7.25). MadCam1 expression was reduced in spleens of WT-LTβR-Ig-treated, TNFR1−/− Ig-treated, and TNFR1−/− LTβR-Ig-treated mice compared to WT-Ig. MadCam1 expression in WT-LTβR-Ig and TNFR1−/− Ig mLNs was reduced compared to WT-Ig, and MadCam1 expression in TNFR1−/− LTβR-Ig mLNs was reduced compared to TNFR1−/−-Ig.

Figure 4

doi: https://doi.org/10.1371/journal.ppat.1002867.g004