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Titolo:
Multiparameter flow cytometric approach for simultaneous evaluation of T lymphocyte-endothelial cell interactions
Autore:
Krupnick, AS; Kreisel, D; Szeto, WY; Popma, SH; Amin, KM; Moore, JS; Rosengard, BR;
Indirizzi:
Univ Penn, Dept Surg, Div Cardiothorac Surg, Med Ctr, Philadelphia, PA 19104 USA Univ Penn Philadelphia PA USA 19104 , Med Ctr, Philadelphia, PA 19104 USA Univ Penn, Med Ctr, Dept Pathol & Lab Med, Philadelphia, PA 19104 USA UnivPenn Philadelphia PA USA 19104 & Lab Med, Philadelphia, PA 19104 USA Univ Penn, Med Ctr, Canc Ctr Flow Cytometry & Cell Sorting Facil, Philadelphia, PA 19104 USA Univ Penn Philadelphia PA USA 19104 ing Facil, Philadelphia, PA 19104 USA
Titolo Testata:
CYTOMETRY
fascicolo: 5, volume: 46, anno: 2001,
pagine: 271 - 280
SICI:
0196-4763(20011015)46:5<271:MFCAFS>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
VASCULAR ENDOTHELIUM; CLONAL EXPANSION; EXPRESSION; APOPTOSIS; INDUCTION; CYCLE; ARTERIOSCLEROSIS; ACTIVATION; ALLOGRAFTS; SECRETION;
Keywords:
endothelial cells; mouse; 5,6-carboxyfluorescein diacetate succinimidyl ester; cytometry;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Rosengard, BR Univ Penn, Dept Surg, Div Cardiothorac Surg, Med Ctr, 6 Silverstein,3400 Spruce, Philadelphia, PA 19104 USA Univ Penn 6 Silverstein,3400 Spruce Philadelphia PA USA 19104
Citazione:
A.S. Krupnick et al., "Multiparameter flow cytometric approach for simultaneous evaluation of T lymphocyte-endothelial cell interactions", CYTOMETRY, 46(5), 2001, pp. 271-280

Abstract

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-T 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(+) or CD4(+) Tcells labeled with 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) were added to a murine endothelial cell monolayer, in which endothelialproliferation was not inhibited by irradiation or addition of a cell cycle-blocking agent. At specific time points, the coculture was analyzed by flow cytometry. T-cell proliferation could be detected by gating on the T-cellsubset and evaluating the CFSE fluorescence peaks. By directly analyzing cellular division, we minimized erroneous interpretation of the data encountered by previous studies, which utilized H-3-thymidine incorporation as sole measure of proliferation. Further subgating on cells that divided facilitated the study of CD8(+) 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-mediated 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. (C) 2001 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/12/20 alle ore 15:22:55