{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,10,16]],"date-time":"2023-10-16T07:18:32Z","timestamp":1697440712434},"reference-count":40,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2001,10,26]],"date-time":"2001-10-26T00:00:00Z","timestamp":1004054400000},"content-version":"vor","delay-in-days":11,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry"],"published-print":{"date-parts":[[2001,10,15]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Since vascular endothelium is now recognized as an immunologically active tissue, a better understanding of the relationship between endothelial cells and T lymphocytes is critical to the field of solid organ transplantation. Investigations of endothelial cell\u2010T cell interactions have been limited by methodology. We developed a flow cytometric method allowing for concurrent investigation of multiple cell populations within the same culture that can be applied to these complex interactions. Allogeneic CD8<jats:sup>+<\/jats:sup> or CD4<jats:sup>+<\/jats:sup> T cells labeled with 5,6\u2010carboxyfluorescein diacetate succinimidyl ester (CFSE) were added to a murine endothelial cell monolayer, in which endothelial proliferation was not inhibited by irradiation or addition of a cell cycle\u2010blocking agent. At specific time points, the coculture was analyzed by flow cytometry. T\u2010cell proliferation could be detected by gating on the T\u2010cell subset and evaluating the CFSE fluorescence peaks. By directly analyzing cellular division, we minimized erroneous interpretation of the data encountered by previous studies, which utilized <jats:sup>3<\/jats:sup>H\u2010thymidine incorporation as sole measure of proliferation. Further subgating on cells that divided facilitated the study of CD8<jats:sup>+<\/jats:sup> lymphocyte activation, differentiation, and acquisition of effector function. By gating on the endothelial cell population, phenotypic changes such as upregulation of surface MHC molecules or immune\u2010mediated apoptosis could be detected. In conclusion, we present a flow cytometric approach that could have important applications for clinical immunological monitoring in allogeneic or xenogeneic transplantation, and might provide the requisite information to better tailor immunotherapy to prevent chronic rejection. Cytometry (Comm. Clin. Cytometry) 46:271\u2013280, 2001. \u00a9 2001 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/cyto.1168","type":"journal-article","created":{"date-parts":[[2002,8,25]],"date-time":"2002-08-25T21:17:12Z","timestamp":1030310232000},"page":"271-280","source":"Crossref","is-referenced-by-count":9,"title":["Multiparameter flow cytometric approach for simultaneous evaluation of T lymphocyte\u2010endothelial cell interactions"],"prefix":"10.1002","volume":"46","author":[{"given":"Alexander S.","family":"Krupnick","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Kreisel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wilson Y.","family":"Szeto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sicco H.","family":"Popma","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kunjlata M.","family":"Amin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonni S.","family":"Moore","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bruce R.","family":"Rosengard","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2002,12,26]]},"reference":[{"key":"e_1_2_5_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/3982"},{"key":"e_1_2_5_3_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.76.4.827"},{"key":"e_1_2_5_4_2","volume-title":"Clinical transplants","author":"Terasaki P","year":"1987"},{"key":"e_1_2_5_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(01)00119-4"},{"key":"e_1_2_5_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/35003221"},{"key":"e_1_2_5_7_2","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199602150-00001"},{"key":"e_1_2_5_8_2","first-page":"325","article-title":"Transplant risks","author":"Takemoto SK","year":"1999","journal-title":"Clin Transplants"},{"key":"e_1_2_5_9_2","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-198207000-00005"},{"key":"e_1_2_5_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.153.3735.545"},{"key":"e_1_2_5_11_2","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199505000-00011"},{"key":"e_1_2_5_12_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.92.2.205"},{"key":"e_1_2_5_13_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.165.8.4182"},{"key":"e_1_2_5_14_2","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-200006270-00019"},{"key":"e_1_2_5_15_2","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.2000.2410"},{"key":"e_1_2_5_16_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.188.9.1751"},{"key":"e_1_2_5_17_2","doi-asserted-by":"publisher","DOI":"10.1006\/cimm.2000.1638"},{"key":"e_1_2_5_18_2","doi-asserted-by":"publisher","DOI":"10.1053\/paor.1999.0049"},{"key":"e_1_2_5_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(94)90236-4"},{"key":"e_1_2_5_20_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI119873"},{"key":"e_1_2_5_21_2","doi-asserted-by":"crossref","first-page":"5212","DOI":"10.4049\/jimmunol.162.9.5212","article-title":"Dynamics and requirements of t cell clonal expansion in vivo at the single\u2010cell level: effector function is linked to proliferative capacity","volume":"162","author":"Gudmundsdottir H","year":"1999","journal-title":"J Immunol"},{"key":"e_1_2_5_22_2","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199905150-00015"},{"key":"e_1_2_5_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(97)00089-6"},{"key":"e_1_2_5_24_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0320(19980615)34:3<143::AID-CYTO4>3.0.CO;2-I"},{"key":"e_1_2_5_25_2","doi-asserted-by":"publisher","DOI":"10.1002\/1097-0320(20001015)42:5<277::AID-CYTO4>3.0.CO;2-A"},{"key":"e_1_2_5_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(01)00371-4"},{"key":"e_1_2_5_27_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.163.6.3131"},{"key":"e_1_2_5_28_2","doi-asserted-by":"publisher","DOI":"10.1002\/eji.1830031011"},{"key":"e_1_2_5_29_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.179.4.1349"},{"key":"e_1_2_5_30_2","doi-asserted-by":"crossref","first-page":"3919","DOI":"10.4049\/jimmunol.156.10.3919","article-title":"Protective immunity in balb\/c mice against the simian virus 40\u2010induced mksa tumor resulting from injection of recombinant large T antigen. 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