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Journal of Virology, October 2008, p. 9978-9993, Vol. 82, No. 20
0022-538X/08/$08.00+0     doi:10.1128/JVI.01326-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

CD20, CD3, and CD40 Ligand Microclusters Segregate Three-Dimensionally In Vivo at B-Cell-T-Cell Immunological Synapses after Viral Immunity in Primate Brain{triangledown}

Carlos Barcia,1,2 Aurora Gomez,1,2 Vicente de Pablos,1,2 Emiliano Fernández-Villalba,1,2 Chunyan Liu,3 Kurt M. Kroeger,3 Javier Martín,1 Andrés Fernández Barreiro,1,2 Maria G. Castro,3,4 Pedro R. Lowenstein,3,4,{dagger} and Maria-Trinidad Herrero1,2*,{dagger}

Clinical and Experimental Neuroscience,1 Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas, School of Medicine, University of Murcia, Campus de Espinardo, 30100 Murcia, Spain,2 Board of Governors’ Gene Therapeutics Research Institute, Cedars-Sinai Medical Center, Los Angeles, California 90048,3 Department of Medicine, and Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 900954

Received 25 June 2008/ Accepted 28 July 2008

The clearance of virally infected cells from the brain is mediated by T cells that engage antigen-presenting cells to form supramolecular activation clusters at the immunological synapse. However, after clearance, the T cells persist at the infection site and remain activated locally. In the present work the long-term interactions of immune cells in brains of monkeys were imaged in situ 9 months after the viral inoculation. After viral immunity, the persistent infiltration of T cells and B cells was observed at the infection sites. T cells showed evidence of T-cell receptor signaling as a result of contacts with B cells. Three-dimensional analysis of B-cell-T-cell synapses showed clusters of CD3 in T cells and the segregation of CD20 in B cells, involving the recruitment of CD40 ligand at the interface. These results demonstrate that immunological synapses between B cells and T cells forming three-dimensional microclusters occur in vivo in the central nervous system and suggest that these interactions may be involved in the lymphocyte activation after viral immunity at the original infection site.

* Corresponding author. Mailing address: Clinical and Experimental Neuroscience, CIBERNED, School of Medicine, University of Murcia, Campus de Espinardo, 30100 Murcia, Spain. Phone: 34 968 36 46 83. Fax: 34 968 36 41 50. E-mail: mtherrer{at}um.es

{triangledown} Published ahead of print on 6 August 2008.

{dagger} P. R. Lowenstein and M. T. Herrero contributed equally to this work.

Journal of Virology, October 2008, p. 9978-9993, Vol. 82, No. 20
0022-538X/08/$08.00+0     doi:10.1128/JVI.01326-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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