Automated technique to evaluate thymocyte adhesion to thymic epithelial cells.
Thymocyte adhesion to thymic epithelial cells is a relevant issue during intrathymic T-cell differentiation, and directly intervenes in the generation and expansion of the T-cell repertoire. In view of these data, it was apparent the usefulness of an automated strategy to evaluate the degree of thymic epithelial cell-thymocyte adhesion. This prompted us to develop an ELISA procedure (using an anti-Thy1 reagent) to determine the degree of thymocyte adhesion onto cultured thymic epithelial cells. The procedure described herein is simple, non-radioactive and reproducible. Additionally, it can potentially be applied to quantitate the degree of thymocyte adhesion to any cellular or non-cellular substrate (for example, extracellular matrix). Moreover, it detected fluctuations of thymocyte adhesion secondary to glucocorticoid treatment of epithelial cells. Thus, it can be regarded as a further tool to analyze intrathymic interactions.
Department of Immunology, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
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Cirne-Lima
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Savino
D000818:Animals; D001331:Automation; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000295:instrumentation; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D018655:Lymphocyte Count; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D013601:T-Lymphocytes / Q000166:cytology* / Q000187:drug effects; D013950:Thymus Gland / Q000166:cytology*