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Isolation, phenotype, and allostimulatory activity of mouse liver dendritic cells

Woo, J and Lu, L and Rao, AS and Li, Y and Subbotin, V and Starzl, TE and Thomson, AW (1994) Isolation, phenotype, and allostimulatory activity of mouse liver dendritic cells. Transplantation, 58 (4). 484 - 491. ISSN 0041-1337

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Donor liver-derived dendritic cells (DC) have recently been identified within various lymphoid and nonlymphoid tissues of organ allograft recipients, including nonimmunosuppressed mice transplanted with and permanently accepting major histocompatibility complex (MHC)-disparate hepatic allografts. These findings have raised questions about the basis of the tolerogenicity of the liver—and, in particular, about the properties of liver-derived DC. To study further the structure, immunophenotype and allostimu-latory activity of leukocytes resident in normal mouse (B10.BR; H-2k, I-Ek) liver, a procedure was developed to maximize the yield of viable, nonparenchymal cells (NPC) obtained following collagenase digestion of perfused liver fragments and density centrifugation (Per-coll). These cells comprised populations expressing lymphoid and myeloid cell surface antigens. As compared with spleen cells, they proved good allostimula-tors of naive (BIO; H-2b, I-E") splenic T cells when tested in primary mixed leukocyte reactions (MLR). After overnight (18-hr) incubation of the NPC, enrichment for transiently adherent, low-density (LD) cells on metrizamide gradients permitted the recovery of low numbers of cells (approx. 2-5 × 105 per liver), many of which displayed distinct DC morphology. Flow cytometric analysis revealed that these cells were CD3-, CD4-, CD8-, and B220-, but strongly expressed CD45 (leukocyte-common antigen), and mild-to-moderate levels of CD lib, heat-stable antigen, and CD44. The cells also expressed moderate intensity of NLDC 145 but not 33D1, DC restricted markers which have been shown to be differentially expressed on mouse DC isolated from various organs. This DC-enriched population was more strongly MHC class II(I-Ek)+ than NPC, as determined by immunocytochemistry and flow cytometry and exhibited much more potent allo-stimulatory activity for naive T cells. These findings demonstrate that freshly isolated murine liver NPC, and perhaps their counterparts in situ, exhibit allo-stimulatory activity that is enhanced in the nonadherent, low-density (DC-enriched) fraction after overnight culture. They further suggest that the © 1994 by Williams and Wilkins.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Woo, J
Lu, L
Rao, AS
Li, Y
Subbotin, V
Starzl, TEtes11@pitt.eduTES11
Thomson, AW
Centers: Other Centers, Institutes, Offices, or Units > Thomas E. Starzl Transplantation Institute
Date: 27 August 1994
Date Type: Publication
Journal or Publication Title: Transplantation
Volume: 58
Number: 4
Page Range: 484 - 491
DOI or Unique Handle: 10.1097/00007890-199408270-00015
Institution: University of Pittsburgh
Refereed: Yes
ISSN: 0041-1337
Other ID: uls-drl:31735062125046, Starzl CV No. 1659
Date Deposited: 08 Apr 2010 17:28
Last Modified: 11 Jun 2021 22:55


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