Fig. 6. Detection of integrin _E receptor (CD103) on human thymocyte subpopulations. (A) DN, DP and SP thymocyte subpopulations were isolated by magnetic cell sorting and subsequently labelled with the antibody against CD103. No expression of CD103 was observed in the DP thymocyte cell population, and only a marginal expression was seen in the CD4⁺ SP population. 12% of the DN subpopulation expressed CD103. The highest expression, however, was observed in the CD8⁺ SP population, in which more than one third of this subpopulation expressed CD103. (B) MACS-isolated CD4^- CD8^- DN thymocytes were triple-stained with antibodies against CD103 (FITC), CD25 (APC), and TCR /ß (PE) or TCR / (PE), respectively. The CD103⁺ DN thymocytes did not show any significant expression of TCR /ß or CD25. A subpopulation of CD103⁺ DN thymocytes expressed the TCR /, the majority of the CD103⁺ DN cells, however, did not express TCR /. (C) Isolated CD8⁺ SP thymocytes were analysed for TCR expression. By one round of magnetic cell sorting, a purity of >92% of CD8⁺ cells was achieved. 4.3% of the CD8⁺ cells expressed the / TCR, whereas 89% of these cells expressed the /ß TCR. By double staining of CD8⁺ cells it was shown that CD103⁺ CD8⁺ cells mostly expressed the /ß TCR, but not the / TCR. (D) Four-colour flow cytometric analysis was performed with MACS-sorted CD8⁺ SP thymocytes, which were labelled with antibodies against CD103 (FITC), CD24 (PE), CD62L (Cy5) and CD69 (APC). All CD103⁺ CD8⁺ SP thymocytes expressed CD69, but not CD24. When the CD103⁺ CD8⁺ cells were gated it could be demonstrated that all of these cells express L-selectin (CD62L) and CD69 simultaneously.

Return to article