行政院國家科學委員會專題研究計畫成果報告:犬傳染性花柳性腫瘤細胞所分泌之毒殺樹枝狀細胞物質之分離及特性分析
Date Issued
2005
Date
2005
Author(s)
朱瑞民
DOI
932313B002104
Abstract
For therapeutic purposes, large numbers of dendritic cells (DC) are essential. However,
the isolation efficiency of canine DC was low. In this study, we established a modified
adherence step to reliably enrich CD14+ monocytes from peripheral blood mononuclear
cells (PBMC). Canine DC were generated from the adherent monocytes. Adherent
monocytes were cultured for 6 days in human GM-CSF, Flt3L, and canine IL-4. This
improved procedure yielded 2–4 times more canine DC than other published methods.
Canine DC characteristics, including surface phenotype and biological functions during
maturation, were also investigated in this study. In canine immature DC (iDC), DLA
class II, CD1a, CD11c, CD40, and CD86 expression was high, based on flow cytometry
and RT-PCR assays. During maturation, which was stimulated by LPS, CD80 and
CD83 were up-regulated in mDC. However, DLA class II, CD1a, CD11c, and CD40
did not increase in mature DC (mDC). Functional maturation was assessed by antigen
uptake ability and the allogenic MLR. Our data showed that incubating canine iDC
with LPS decreased antigen uptake and increased the immunostimulatory capacity,
indicating that LPS accelerates DC maturation. The TNF-a gene expression was
decreased during DC differentiation from monocytes confirm the functional difference
between monocytes and DC. This new protocol markedly increased the cell number
with in vitro culture of DC precursors and thus may facilitate the use of DC in clinical
cellular immunotherapy.
Publisher
臺北市：國立臺灣大學獸醫學系暨研究所
Type
report
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