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Titolo:
INTERFERON-GAMMA-SECRETING T-CELL POPULATIONS IN REJECTING MURINE CARDIAC ALLOGRAFTS - ASSESSMENT BY FLOW-CYTOMETRY
Autore:
STINN JL; TAYLOR MK; BECKER G; NAGANO H; HASEGAWA S; FURAKAWA Y; SHIMIZU K; LIBBY P; MITCHELL RN;
Indirizzi:
BRIGHAM & WOMENS HOSP,DEPT PATHOL,DIV IMMUNOL,LMRC 515,221 LONGWOOD AVE BOSTON MA 02115 BRIGHAM & WOMENS HOSP,DEPT PATHOL,DIV IMMUNOL BOSTON MA 02115 BRIGHAM & WOMENS HOSP,DEPT MED BOSTON MA 02115 OSAKA UNIV,SCH MED,DEPT SURG OSAKA JAPAN TOKYO MED & DENT UNIV,DEPT CARDIOTHORAC SURG TOKYO 113 JAPAN
Titolo Testata:
The American journal of pathology
fascicolo: 5, volume: 153, anno: 1998,
pagine: 1383 - 1392
SICI:
0002-9440(1998)153:5<1383:ITPIRM>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE CHAIN-REACTION; CYTOKINE PRODUCTION; IN-VIVO; EXPRESSION; ACTIVATION; MOUSE; ARTERIOSCLEROSIS; MECHANISMS; T-HELPER-1; SUBSETS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
32
Recensione:
Indirizzi per estratti:
Citazione:
J.L. Stinn et al., "INTERFERON-GAMMA-SECRETING T-CELL POPULATIONS IN REJECTING MURINE CARDIAC ALLOGRAFTS - ASSESSMENT BY FLOW-CYTOMETRY", The American journal of pathology, 153(5), 1998, pp. 1383-1392

Abstract

Interplay between T-helper-1 (Th1) and T-helper-2 (Th2) cells is considered important in the development of acute allograft rejection and many other immune-mediated disease processes. Existing methods for evaluating expression of Th1 and Th2 cytokines, including reverse transcriptase polymerase chain reaction (RT-PCR), RNase protection assay (RPA), immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA) all have limitations; alternate techniques to quantify cell populations expressing specific cytokine proteins, generate statistically analyzable data, and allow simultaneous identification of cytokine-secretingcell type are needed. To this end, we adapted a now cytometric technique for intracellular cytokine immunofluorescence staining for use with cells belated from solid tissue. To demonstrate the utility of the method, we determined the number of CD4(+) and CD8(+) cells secreting the prototypical Th1 and Th2 cytokines, interferon (IFN)-gamma, and interleukin (IL)-4 in acutely rejecting murine cardiac allografts. We also measured the cytokine production via ELISA, RPA, and semiquantitative competitive RT-PCR. The number of CD4+ cells producing IFN-gamma increased as rejection proceeded, in agreement with previous data; we detected no IL-4 production at any time, although relatively low numbers of IL-10-producing cells were identified. In addition, a high percentage of CD8+ cells, which outnumber CD4(+) cells at day 6 after transplant, also produce IFN-gamma, suggesting that cytotoxic lymphocytes contribute significantly to the local cytokine milieu. This new application of intracellular cytokine staining provides a powerful methodology for studying transplantation immunology. The method may also be easily adapted to the study of other immune-mediated processes.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 09/07/20 alle ore 23:09:01