The Journal of Experimental Medicine
Keystone Symposia
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Published online January 29, 2007
doi:10.1084/jem.20061890
The Journal of Experimental Medicine, Vol. 204, No. 2, 345-356
The Rockefeller University Press, 0022-1007 $30.00
© 2007 Boissonnas et al.
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ARTICLE

 In vivo imaging of cytotoxic T cell infiltration and elimination of a solid tumor

Alexandre Boissonnas1, Luc Fetler2, Ingrid S. Zeelenberg1, Stéphanie Hugues1, and Sebastian Amigorena1

1 Institut National de la Santé et de la Recherche Médicale U653, Immunité et Cancer, Pavillon Pasteur, Institut Curie, F-75245 Paris Cedex 05, France
2 Centre National de la Recherche Scientifique UMR 168, Laboratoire de Physico-Chimie Curie, Institut Curie, F-75245 Paris Cedex 05, France

CORRESPONDENCE Sebastian Amigorena: sebas{at}curie.fr

Although the immune system evolved to fight infections, it may also attack and destroy solid tumors. In most cases, tumor rejection is initiated by CD8+ cytotoxic T lymphocytes (CTLs), which infiltrate solid tumors, recognize tumor antigens, and kill tumor cells. We use a combination of two-photon intravital microscopy and immunofluorescence on ordered sequential sections to analyze the infiltration and destruction of solid tumors by CTLs. We show that in the periphery of a thymoma growing subcutaneously, activated CTLs migrate with high instantaneous velocities. The CTLs arrest in close contact to tumor cells expressing their cognate antigen. In regions where most tumor cells are dead, CTLs resume migration, sometimes following collagen fibers or blood vessels. CTLs migrating along blood vessels preferentially adopt an elongated morphology. CTLs also infiltrate tumors in depth, but only when the tumor cells express the cognate CTL antigen. In tumors that do not express the cognate antigen, CTL infiltration is restricted to peripheral regions, and lymphocytes neither stop moving nor kill tumor cells. Antigen expression by tumor cells therefore determines both CTL motility within the tumor and profound tumor infiltration.

Abbreviations used: CFP, cyan fluorescent protein; PI, propidium iodide; SHG, second harmonic generation; TPLSM, two-photon laser-scanning microscopy.

A. Boissonnas and L. Fetler contributed equally to this work.


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