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Journal of Histochemistry and Cytochemistry, Vol. 45, 1293-1298, Copyright © 1997 by The Histochemical Society, Inc.

TECHNICAL NOTE

Murine Thymic Nurse Cells and Rosettes: Analysis of Adhesion Molecule Expression Using Confocal Microscopy and a Simplified Enrichment Method

Antonio J. Oliveira-dos-Santosa, Theresa Rieker-Geleya, Heidrun Recheisa, and Georg Wicka
a Institute for General and Experimental Pathology, Medical School, University of Innsbruck, Innsbruck, Austria

Correspondence to: Georg Wick, Inst for General and Experimental Pathology, U. of Innsbruck, Fritz-Pregl-Str. 3/IV, Innsbruck A-6020, Austria.

Thymic nurse cells (TNC) and T-cell stromal rosettes (ROS) are two in vivo models for stromal cell-thymocyte interactions. We describe a simplified enrichment method for TNC and ROS that overcomes the necessity for large amounts of tissue. The complexes were further analyzed with confocal microscopy, and three subunits of ROS were defined on the basis of their central cell phenotype, i.e., macrophage, dendritic, or epithelial cell rosettes. Because adhesion molecules are proposed to play a crucial role in T-cell development, we investigated CD44, LFA-1, and ICAM-1 expression in such complexes. The epithelial component of TNC expresses CD44 and ICAM-1, whereas intra-TNC thymocytes are LFA-negative. With regard to ROS, all subsets expressed CD44, and macrophage and dendritic cell ROS were also ICAM-1-positive and LFA-1-positive. The current protocol opens the possibility for further in vivo analysis of stromal cell-thymocyte interactions, e.g., for studies of scarce gene mutant mice. (J Histochem Cytochem 45:1293-1297, 1997)

Key Words: thymic nurse cell, thymic rosette, thymocyte, confocal microscopy


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