Subtractive isolation of phage-displayed single-chain antibodies to thymic stromal cells by using intact thymic fragments
Proceedings of the National Academy of Sciences of the United States of America , Volume 94 - Issue 8 p. 3903- 3908
In the murine thymus, the stroma forms microenvironments that control different steps in T cell development. To study the architecture of such microenvironments and more particularly the nature of communicative signals in lympho–stromal interaction during T cell development, we have employed the phage antibody display technology, with the specific aim of isolating thymic stromal cellspecific single-chain antibodies from a semisynthetic phage library. A subtractive approach using intact, mildly fixed thymic fragments as target tissue and lymphocytes as absorber cells generated monoclonal phages (MoPhabs) detecting subsets of murine thymic stromal cells.
In the present paper we report on the reactivity of single-chain antibodies derived from three MoPhabs, TB4–4, TB4–20, and TB4–28. While TB4–4 and TB4–20 are both epithelium specific, TB4–28 detects an epitope expressed on both epithelial- and mesenchymal-derived stromal cells. TB4–4 reacts with all cortical epithelial cells and with other endoderm-derived epithelia, but this reagent leaves the majority of medullary epithelial cells unstained. In contrast, MoPhab TB4–20 detects both cortical and medullary thymic epithelial cells, as well as other endoderm- and ectoderm-derived epithelial cells. Cross-reaction of single-chain antibodies to human thymic stromal cells shows that our semisynthetic phage antibody display library, in combination with the present subtractive approach, permits detection of evolutionary conserved epitopes expressed on subsets of thymic stromal cells.
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|Proceedings of the National Academy of Sciences of the United States of America
|Department of Immunology
van Ewijk, W., de Kruif, J., Germeraad, W., Berendes, P., Röpke, C., Platenburg, P. P., & Logtenberg, T. (1997). Subtractive isolation of phage-displayed single-chain antibodies to thymic stromal cells by using intact thymic fragments. Proceedings of the National Academy of Sciences of the United States of America, 94(8), 3903–3908. doi:10.1073/pnas.94.8.3903