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. Author manuscript; available in PMC: 2022 Feb 18.
Published in final edited form as: Cell. 2021 Jan 27;184(4):1000–1016.e27. doi: 10.1016/j.cell.2020.12.040

Figure 7. Dural sinus-associated APCs present CSF antigens to patrolling T cells.

Figure 7

(A) Immunohistochemistry of Nur77-GFP expression in CD3+ T cells at the superior sagittal sinus of Nur77-GFP mice, representative of n=3 mice.

(B, C) Flow cytometry and quantification of Nur77-GFP expression in CD4 and CD8 T cells in the dura of Nur77-GFP mice, n=4 mice.

(D) Representative two-photon microscopy snapshot and tracers of T cell trajectories over a two hour imaging period at the superior sagittal sinus of RAG2 KO mice 14 days post transfer of UBC-GFP CD4 T cells. Tracers are color coded by the length of the tracing period (0–120 minutes). S.S. sinus=superior sagittal sinus.

(E–G) Quantification of individual T cell total displacement per hour (μm/hr), average velocity (μm/min), and T cell arrest coefficient (fraction of time velocity<0.2 μm/min) during 60–120 minutes of two-photon microscopy. Color coded based on T cells from individual mice, n=3 mice, n=207 individual T cells tracked total.

(H–L) Flow cytometry and quantification of dural Thy1.2– CD4 T cells or Thy1.2+ TCRVβ5.1/2+ OT-II CD4 T cell numbers and phenotype in the dural meninges of Thy1.1 mice three or 14 days following PBS or OVA323–339 i.c.m. administration and i.v. transfer of DC-activated OT-II cells. Activated=CD44+CD69–, tissue resident memory (TRM)=CD44+CD69+.* p<0.05 (Student’s t-test), n=5 mice/group.

(M, N) Immunohistochemistry and quantification of CD3 coverage at non-sinus, superior sagittal sinus, and transverse sinus regions of naïve mice or three-14 days post experimental autoimmune encephalomyelitis (EAE) induction. No stars=not significant, * p<0.05, ** p<0.01, *** p<0.001 vs naïve mice (One-way ANOVA with Dunnett’s post-hoc test) n=4–5 mice/group. T. sinus=transverse sinus, S.S. sinus=superior sagittal sinus.

(O–Q) Flow cytometry and quantification of Thy1.2− CD4 T cells or Thy1.2+ TCRVβ11+ 2D2 CD4 T cell numbers and phenotype in the dural meninges of naïve or day 14 EAE induced Thy1.1 mice after i.v. transfer of naïve 2D2 T cells at day 7. Activated=CD44+CD69−, tissue resident memory (TRM)=CD44+CD69+.** p<0.01 (Student’s t-test), n=5 mice/group.

See also Figure S8.