{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,3]],"date-time":"2026-02-03T01:50:14Z","timestamp":1770083414530,"version":"3.49.0"},"reference-count":23,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2001,3,1]],"date-time":"2001-03-01T00:00:00Z","timestamp":983404800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2001,3,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>We have previously shown that human endothelial cells (EC) are less efficient than professional APC, e.g., B lymphoblastoid cells (BLC), at stimulating allogeneic CD8+ T cells to develop into CTL. In this study we describe FACS-based limiting dilution analyses using the dilution of the intracellular dye CFSE as an indicator of CD8+ T cell alloactivation and expansion with significantly increased sensitivity compared with conventional, cytotoxicity-based assays. In addition, this assay permits the relative size of clonal CTL populations that are generated in individual CD8+ T cell cultures to be determined (clonal burst size). We have applied this method to quantitatively compare the generation of CTL at the clonal level following stimulation of allogeneic CD8+ T cells by either BLC or HUVEC derived from the same donor. CD8+ T cells expanded by allostimulation were identified as CD8+, CFSElow cells and were categorized as CTL by the expression of intracellular perforin and IFN-\u03b3. Precursor frequencies for EC-stimulated CTL were 5- to 40-fold (mean, 7.5-fold) lower compared with BLC-stimulated CTL (p &amp;lt; 0.01). Concomitantly, the average clonal burst sizes in EC-stimulated CTL cultures were significantly smaller than those in conventional CTL cultures, primarily due to the occurrence of some very large clone sizes exclusively with BLC stimulation. Although EC-stimulated CTL were generated only from the memory subset of CD8+ T cells, BLC-stimulated very large burst sizes of CTL were observed from both naive and memory CD8+ T cell precursors. These data establish that both a lower frequency of reactive precursors and more limited clonal expansion, but not regulatory T cells, contribute to the reduced capacity of EC to promote alloreactive CTL differentiation compared with that of professional APC.<\/jats:p>","DOI":"10.4049\/jimmunol.166.6.3846","type":"journal-article","created":{"date-parts":[[2014,4,20]],"date-time":"2014-04-20T19:32:32Z","timestamp":1398022352000},"page":"3846-3854","source":"Crossref","is-referenced-by-count":27,"title":["Human Vascular Endothelial Cells Stimulate a Lower Frequency of Alloreactive CD8+ Pre-CTL and Induce Less Clonal Expansion than Matching B Lymphoblastoid Cells: Development of a Novel Limiting Dilution Analysis Method Based on CFSE Labeling of Lymphocytes"],"prefix":"10.1093","volume":"166","author":[{"given":"Thomas J","family":"Dengler","sequence":"first","affiliation":[{"name":"Interdepartmental Program in Vascular Biology and Transplantation, Boyer Center for Molecular Medicine, Yale University School of Medicine , New Haven, CT 06510"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David R","family":"Johnson","sequence":"additional","affiliation":[{"name":"Interdepartmental Program in Vascular Biology and Transplantation, Boyer Center for Molecular Medicine, Yale University School of Medicine , New Haven, CT 06510"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jordan S","family":"Pober","sequence":"additional","affiliation":[{"name":"Interdepartmental Program in Vascular Biology and Transplantation, Boyer Center for Molecular Medicine, Yale University School of Medicine , New Haven, CT 06510"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2001,3,15]]},"reference":[{"key":"2026020208085001800_R1","first-page":"403","volume-title":"Transplantation","year":"1996"},{"key":"2026020208085001800_R2","first-page":"285","volume-title":"Clin. 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