Clonal Expansions of CD8+ T Cells Dominate the T Cell Infiltrate in Active Multiple Sclerosis Lesions as Shown by Micromanipulation and Single Cell Polymerase Chain Reaction

doi:10.1084/jem.192.3.393

Published online 8 August 2000.

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© The Rockefeller University Press, 0022-1007/2000/8/393/ $5.00
The Journal of Experimental Medicine, Volume 192, Number 3, August 7, 2000 393-404

Original Article

Clonal Expansions of CD8⁺ T Cells Dominate the T Cell Infiltrate in Active Multiple Sclerosis Lesions as Shown by Micromanipulation and Single Cell Polymerase Chain Reaction

Holger Babbe^a, Axel Roers^a, Ari Waisman^a, Hans Lassmann^c, Norbert Goebels^d, Reinhard Hohlfeld^d, Michael Friese^b, Roland Schröder^b, Martina Deckert^e, Stephan Schmidt^f, Rivka Ravid^g, and Klaus Rajewsky^a
^a Institute for Genetics, Institute for Pathology, University of Cologne, 50931 Cologne, Germany
^b Department of Neuropathology, Institute for Pathology, University of Cologne, 50931 Cologne, Germany
^c Department of Neuroimmunology, Brain Research Institute, University of Vienna, 1090 Vienna, Austria
^d Department of Neuroimmunology, Max-Planck-Institute for Neurobiology, 82152 Martinsried, Germany
^e Department of Neuropathology, University of Bonn, 53105 Bonn, Germany
^f Department of Neurology, University of Bonn, 53105 Bonn, Germany
^g Netherlands Brain Bank, 1105 AZ Amsterdam, The Netherlands

Correspondence to: Axel Roers, Department of Immunology, Institute for Genetics, University of Cologne, Weyertal 121, 50931 Cologne, Germany. Tel:49-221-478-4543 Fax:49-221-478-6360 E-mail:axel.roers{at}uni-koeln.de.

Clonal composition and T cell receptor (TCR) repertoire of CD4⁺and CD8⁺ T cells infiltrating actively demyelinating multiplesclerosis (MS) lesions were determined with unprecedented resolutionat the level of single cells. Individual CD4⁺ or CD8⁺ T cellswere isolated from frozen sections of lesional tissue by micromanipulationand subjected to single target amplification of TCR-ßgene rearrangements. This strategy allows the assignment ofa TCR variable region (V region) sequence to the particularT cell from which it was amplified. Sequence analysis revealedthat in both cases investigated, the majority of CD8⁺ T cellsbelonged to few clones. One of these clones accounted for 35%of CD8⁺ T cells in case 1. V region sequence comparison revealedsigns of selection for common peptide specificities for someof the CD8⁺ T cells in case 1. In both cases, the CD4⁺ T cellpopulation was more heterogeneous. Most CD4⁺ and CD8⁺ cloneswere represented in perivascular infiltrates as well as amongparenchymal T cells. In case 2, two of the CD8⁺ clones identifiedin brain tissue were also detected in peripheral blood. Investigationof the antigenic specificities of expanded clones may help toelucidate their functional properties.

Key Words: autoimmunity, demyelinating disease, T cell receptor ßchain, gene rearrangement, peripheral blood

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