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Isolation and characterization of high endothelial cell lines derived from mouse lymph nodes

  • Immunology
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Summary

Two long-term cultured cell lines were established from BALB/c mouse axillary and cervical lymph nodes that exhibited a combination of functional, morphological, and phenotypic characteristics consistent only with high endothelial venule cells. Spleen lymphocytes selectively bound and migrated across the cell lines. On Matrigel, these cell lines formed tubules with lumens, a characteristic unique to endothelial cells. Morphologically the cells were 20–30 μm in diameter and exhibited contact inhibition. The cells were not myeloid in origin because they lacked sodium fluoride-inhibitable nonspecific esterase activity, myeloperoxidase activity, and F4/80 antigen. The cell line phenotypes were compared to high endothelial venule (HEV) cells in tissue sections. HEV cells in lymph node tissue sections expressed endoglin, PECAM-1, ICAM-1, VCAM-1, laminin, fibronectin, collagen IV, H2Kd, MECA 79, MECA 325, and vWF. The cell lines expressed endoglin, VCAM-1, fibronectin, and H2Kd. The cell line derived from cervical lymph nodes also expressed laminin and H2Dd. Neither cell line expressed collagen IV, IAd, ICAM-1, ICAM-2, dendritic cell antigen, or PECAM-1. They also did not express MECA antigens or intracellular vWF, consistent with reports of many cultured endothelial cells. To further substantiate cell line identification, antiserum generated against the cell lines bound specifically to HEV cells in frozen lymph node tissue sections and to both of the lymph node-derived cell lines but not control cell lines. Thus, the lymph node derived-cell lines expressed molecules found on HEV cellsin vivo and most importantly retained the functions of tubule formation, lymphocyte adhesion, and promotion of lymphocyte migration.

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  1. Department of Pathology & Laboratory Medicine, University of Cincinnati College of Medicine, 231 Bethesda Avenue, P.O. Box 670529, 45267-0529, Cincinnati, Ohio

    Joan M. Cook-Milis & Joan S. Gallagher

  2. Department of Microbiology & Immunology, Northwestern University Medical School, 60611, Chicago, Illinois

    Joan M. Cook-Milis & Thomas L. Feldbush

  3. Veterans Administration Lakeside, 60611, Chicago, Illinois

    Joan M. Cook-Milis

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Cook-Milis, J.M., Gallagher, J.S. & Feldbush, T.L. Isolation and characterization of high endothelial cell lines derived from mouse lymph nodes. In Vitro Cell.Dev.Biol.-Animal 32, 167–177 (1996). https://doi.org/10.1007/BF02723682

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