Activation of bone marrow−resident memory T cells by circulating, antigen-bearing dendritic cells

Lois L Cavanagh, Roberto Bonasio, Irina B Mazo, Cornelia Halin, Guiying Cheng, Adrianus W M van der Velden, Annaiah Cariappa, Catherine Chase, Paul Russell, Michael N Starnbach, Pandelakis A Koni, Shiv Pillai, Wolfgang Weninger & Ulrich H von Andrian

Supplementary Video 1 (mov 3M)
Representative 3D time-lapse movie generated by 2-photon intravital microscopy in the skull of an anesthetized mouse. Hoechst 33342-labeled (blue) P14 TCM were injected into recipient mice and allowed to distribute in the body. 18h later, CMTMR-labeled (red) purified spleen DC were injected i.v. without peptide pulsing. Two hours thereafter, the animal was prepared for 2-photon intravital microscopy of skull BM as described in Materials and Methods. The luminal compartment of BM microvessels was delineated by i.v. injection of FITC-dextran (2MDa). 3D image stacks (23 optical sections, 5 m vertical step size) were acquired every minute during a 30 min scan period. The total imaged volume (w x l x d) was 210 m x 210 m x 110 m.

Supplementary Video 2 (mov 3M)
Representative 3D time-lapse movie generated as described for movie 1, except that DC were pulsed with antigenic gp33 peptide prior to injection. Note that most TCM move at reduced velocity and undergo long-lasting tight contacts with DC.

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