A friend recently presented at a journal club on a paper titled "Effector T cell interactions with meningial vascular structures in nascent autoimmune CNS lesions," where the authors used two-photon imaging to create movies of T cells entering the vasculature and parenchyma of rat spinal cords. These rats had experimental autoimmune encephalomyelitis (EAE), and were administered T cells that were 1) engineered to express GFP, and 2) specific for myelin basic protein (MBP), the pathological antigen associated with EAE. The movies are really fun to watch as the T cells traverse the blood vessels epithelial cells, with great use of the two-photon technology.
I've added some videos from the 10 that are available. At the end of the post, there is a link to download the article and these movies ("supplemental" data, they say!) if your interested in learning more about the model and what is going on!
Supp. Movie #1: Compare the number of T cells between 1 and 2.5 days after transfer of the effector T cells. The arrows signify the direction of blood flow.
Supp. Movie #4: Watch T cells pass through vessel walls, complete with side-by-side 3D models!
Supp. Movie #5: This one is really cool. The authors found that two cell adhesion molecules were particularly important for the T cell locomotion and passage through the blood-brain barrier during EAE. Then they use antibodies against the CAMs to "wash away" the T cells. In this video they first pretreat with antibody against one of the CAMs (LFA-1), and then they add antibody against the other (VLA-4).Note the countdown as they add the second antibody!
Supp. Movie #6: My personal favorite. Using Z-stacks and maximum projections, the authors were able to get a huge image of many vessels, allowing them to display the escalation of T cells infiltrating the CNS parenchyma.
Supp. Movie #8: What happens to the T cells once they pass the blood-brain barrier? The find antigen-presenting phagocytes that patrol the perivascular and meningial space. More 3D rendering as well!
Supp. Movie #9: When the perivascular/meningial phagocytes and T cells meet up, they have prolonged contact. The article also mentions that the T cells begin to substantial amounts of inflammatory cytokines, including IFNg, IL-17, and and TNFa.
Hope you enjoyed the videos, and for the article and files, click the link below:
http://www.nature.com/nature/journal/v462/n7269/suppinfo/nature08478.html
Reference:
Ingo Bartholomäus,
Naoto Kawakami,
Francesca Odoardi,
Christian Schläger,
Djordje Miljkovic,
Joachim W. Ellwart,
Wolfgang E. F. Klinkert,
Cassandra Flügel-Koch,
Thomas B. Issekutz,
Hartmut Wekerle
&
Alexander Flügel
Nature 462, 94-98(5 November 2009)
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